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Background: Acinetobacte is the leading cause of pneumonia and sepsis in the ICU ward. Accordingly, in the present study, the antibiotic susceptibility pattern, presence, and dissemination of different classes of integrons and fluoroquinolone resistance genes were investigated among A. baumannii isolates. Methods: In this descriptive, cross-sectional study, during a period of 24 months (2018 - 2020), 100 isolates of A. baumannii were isolated from different clinical specimens of patients admitted to the two teaching hospital in Ardabil province in the northwest of Iran. Kirby -Bauer disk diffusion, PCR, and sequencing methods were used for antimicrobial susceptibility testing and gene and mutation verification. Results: The resistance rates to all tested antibiotics were found to be between 78% and 100%. No isolate was resistant to polymyxin B. Multidrug-resistant (MDR) rate among tested clinical isolates was about 99%. The prevalence of class 1, 2, and 3 integrons was found to be 70%, 21%, and 0%, respectively. The aadA1 cassette gene was detected in all class 1 integron-carrying strains. Conclusions: High-level antibiotic resistance and a high prevalence of integrons were observed among these clinical isolates. Our findings highlighted the need for continuous monitoring of resistant isolates.

Due to the lack of effective vaccine(s) against leishmania and also pharmacokinetics issues of current drugs, it is necessary to discover new antileishmanial agents. Within this particular study, a series of novel 1-aryl/alkyl-3-benzoyl/cyclopropanoyl thiourea derivatives were synthesized (yields 69-84%) and evaluated as antileishmanial compounds (1-11). Synthetic derivatives were subjected to in vitro antileishmanial assessment against Leishmania major promastigotes by colorimetric MTT assay. Compounds 3 (IC

Background Blastocystis is a common intestinal protozoa found in animal and human fecal samples, with over 1 billion individuals infected worldwide. Since domestication, dogs and cats have had a close bond with humans. However, their close proximity poses a potential health risk since they may harbor several zoonotic agents. A global estimate of Blastocystis infection and subtype (ST) distribution in dogs and cats would therefore be of great health importance to humans. Methods We performed a comprehensive systematic search of four English-language databases (PubMed, Scopus, Google Scholar, Web of Science) for relevant articles up to 8 November 2021. The random-effects model was used to make pooled estimates with confidence intervals (CIs). Results In total, we identified 49 publications that met our inclusion criteria and subsequently analyzed the 65 datasets in these articles, of which 23 and 42 datasets were on cats and dogs, respectively. Among the 2934 cats included in the 23 datasets, which involved 16 countries, the prevalence rate of Blastocystis infection was 9.3% (95% CI 5.3–15.9%). The prevalence of Blastocystis infection was slightly lower [7%, 95% CI 4.7–10.4%) among the 7946 dogs included in the 42 datasets, involving 23 countries. The sensitivity analysis showed that no remarkable variation in the estimates upon the stepwise removal of each dataset. Higher ST diversity was found among the examined dogs (ST1-8, ST10, ST23, ST24) than among cats (ST1-4, ST10, ST14). Among dogs, ST3 was the most frequent ST (41.3%), followed by ST2 (39.3%), ST1 (30.9%), ST4 (13.4%), ST8 (12.7%), ST10 (11%) and ST5 (8.1%). Also among dogs, each of ST6, ST7, ST23 and ST24 was observed in only one study. Of the ST found in the cats examined, ST4 (29.5%), followed by ST10 (22.5%), ST1 (19.8%) and ST3 (17.6%) were the most common. A single study also reported the presence of both ST2 and ST14 in cats. With respect to zoonotic Blastocystis STs (ST1–ST9 and ST12), eight were reported from dogs (ST1-ST8) and four were isolated from cats (ST1–ST4), showing the implication of dog and cats in zoonotic transmission. Conclusions Taken together, our results show that elucidation of the true epidemiology and ST distribution of Blastocystis in dogs and cats demands more comprehensive studies, particularly in the negelected regions of the world. Graphical Abstract

Background & objectives: Drug-repurposing is the study on application of existing drugs for treatment or control of other diseases. Major advantage of the technique relies on a nominated drug molecule that is applied for pharmacodynamics optimization due to lack of serious pharmacokinetics challenges. According to the importance of the subject, a present contribution has been dedicated to the in-silico analysis of a few drug classes with the aim of achieving potential anti-leishmanial pharmacophores. Methods: 3D structure of protein targets within leishmania parasite were retrieved from Brookhaven Protein Data Bank (PDB) on the basis of literature reports to evaluate the related complexes with drugs via molecular docking. Qualitative and quantitative analysis of drug-target interaction patterns in docked complexes offered drugs with higher binding affinities toward targets and finally structural patterns or hypothetical anti-leishmanial pharmacophores were proposed with regard to the top-ranked pharmaceutical compounds. Results: Highest free binding energy could be estimated for Nateglinide in binding to farnesyl diphosphate synthase (ΔGb -13.30 kcal/mol). Among steroids, Norgestrel synthase (ΔGb -9.48 kcal/mol) and Testosterone synthase (ΔGb -8.05 kcal/mol) exhibited higher enzyme binding affinities and Arg82 was a key residue in making hydrogen bonds. Within fused tricyclic structural patterns, mirtazapine exhibited highest binding affinity to deoxy uridine triphosphate (ΔGb -8.64 kcal/mol). In Carbamazepine, amide substituent of the central ring facilitated the formation of two effective hydrogen bonds with Gln21 and Asn25 in deoxy uridine triphosphate. Conclusion: On the basis of obtained results for steroids and fused tricyclic scaffolds, it will be possible to design molecules that can inhibit several pathogenic targets simultaneously.

Background and purpose: The prevalence of leishmaniasis is reported in more than 98 countries and Iran is one of the endemic areas. There is no vaccine for this disease and few effective drugs are available to treat it. Moreover, drug resistance to the disease is increasing. During the past decade, several in vitro and in vivo studies have been performed on dihydropyrimidine derivatives as antileishmanial agents. Experimental approach: In the present project, a few 6-methyl-4-aryl-N-aryl dihydropyrimidinone/thiones (A7-A11) and N-heteroaryl-3-(para-methoxy benzyl) amino but-enamides (A1-A6) were synthesized, structurally characterized, and finally subjected to in vitro anti-leishmanial effect against Leishmania major promastigotes. Findings / Results: Results of the study showed that compound A10, 4-(3-chlorophenyl)-6-methyl-N-phenyl- 2-thioxo-1,2,3,4-tetrahydropyrimidine-5-carboxamide, exhibited superior anti-leishmanial effect with IC50 value of 52.67 μg/mL (more active than standard drug Glucantim® with IC50 71000 ± 390 μg/mL). Conclusion and implications: It was demonstrated that some dihydropyrimidine thiones were able to inhibit Leishmania major promastigotes. Structure-activity relationship evaluations indicated that more electron-poor rings such as isoxazole afforded higher activity within A1-A6 series and in these derivatives, N-benzothiazole rings reinforced anti-leishmanial activity concerning thiazole. It was also observed that higher anti-parasite activities of A10 and A11 concerning A7-A9 might be related to the incorporation of the sulfur atom into C2 position, replacement of N-thiazole carboxamide by N-phenyl carboxamide on C5 position of dihydropyrimidine ring, and also replacement of para with meta-substituted phenyls within C4 of dihydropyrimidine ring. The results may help unveil new 4-aryl-5-carboxamide dihydropyrimidines as potential anti-leishmanial agents and their further structural modification toward more potent derivatives.

Sarcocystis species are diverse apicomplexan parasites, though only two zoonotic species (S. hominis and S. heydorni) circulate between cattle and humans. Due to the importance of cattle in the human food chain and to prevent the consequences of parasitosis in humans, the first global systematic review and meta-analysis on molecular epidemiology, species distribution, and zoonotic significance of Sarcocystis infection in cattle was performed.For this aim, four international English databases (PubMed, Scopus, Google Scholar, and Web of Science) were systematically searched till 20th September 2021, and random-effect models were drawn to calculate total estimates and their 95% confidence intervals (CIs).Finally, 44 papers from 21 countries were qualified for this review which examined 8526 cattle regarding Sarcocystis infection, rendering a total prevalence of 62.7% (95% CI 53-71.5%). Globally, 12 Sarcocystis spp. have been reported from cattle, including S. cruzi, S. hominis, S. hirsuta, S. rommeli, S. heydorni, S. bovifelis, S. bovini, S. sinensis, S. gigantea, S. fusiformis, S. hjorti and S. tenella. Among them, S. cruzi (37 studies), S. hominis (22 studies) and S. hirsuta (19 studies) were the 3 most common species, with 76.4% (95% CI 64.8-85%), 30.2% (95% CI 19.3-44%) and 8.7% (95% CI 3.8-18.6%), respectively. However, molecular identification was not performed in 48.4% (95% CI 27.3-70.1%) of the positive samples.Despite the zoonotic significance of Sarcocystis spp., particularly S. hominis, the epidemiology and distribution of Sarcocystis infection in cattle remains unclear and demands more extensive researches around the world.

Background: Cystic Echinococcosis is considered a cosmopolitan cyclozoonotic parasitic infection. This study aimed at evaluating the seroprevalence of human hydatidosis using ELISA test and find the role of mutable factors such as age, sex, occupation, residency in the broadcast of the parasites in rural Ardabil Province, North-West of Iran. Methods: The study population was 950 asymptomatic individuals selected randomly from urban and rural populations of Ardabil province, North-West of Iran by randomized cluster sampling in 2019-2020. Immunoglobulin G antibodies against Echinococcus granulosus spp. were analyzed by ELISA test. Data were analyzed using SPSS software and Multivariable logistic regression model. Results: Overall, 42 (4.4%) of the participants had anti E. granulosus antibodies in this region. High titer antibodies were most prevalent in people age group of >70 yr old, rural areas, females and people having history of contact with dog that showed significant difference. There was no significant association between the presence of Echinococcus antibodies and sex, occupation, having history of eating unwashed vegetable. Conclusion: This is the first description of the seroprevalence of E. granulosus infection in the population in Ardabil Province, North-West of Iran. Obtained rate of hydatidosis approves the importance of diagnosing human cystic echinococcosis in these regions and it is expected that the authorities be careful to screen the disease.

In the microevolutionary scale of Giardia lamblia, the gene migration indicates how G. lamblia assemblages have transmitted between adjacent counties. 33 positive fecal samples were taken from patients suffering gastrointestinal disorders (nausea, bloating, burping constipation and fatty diarrhea) at Tabriz and Ardabil cities, where located in the cold regions of northwest Iran. Following parasitological examinations, DNA samples were extracted, amplified and digested by single-step PCR-RFLP assay, targeting the glutamate dehydrogenase (gdh) locus to distinguish within and between assemblages A and B. PCR products were directly sequenced to reconfirm their heterogeneity traits and phylogenetic analysis. Of the 33 isolates, 81.9% (n: 27), 9% (n: 3) and 9% (n: 3) were successfully identified as assemblages A (genotype AII), B (genotype BIII) and the mixed of genotypes AII and B, respectively. Despite the presence of heterogeneous clinical backgrounds, a low genetic diversity of sub-assemblage AII was identified among symptomatic cases. A low value of pairwise fixation index showed that G. lamblia sub-assemblage AII is not genetically differentiated among northwest regions of Iran. The occurrence of haplotypes TAB-1/ARD-1 between two regional populations indicates that there is a dawn of G. lamblia gene flow due to transfer of alleles through host mobility and/or ecological alterations. To assess the hypothetical evolutionary scenario, further studies are essential for multilocus genotyping of G. lamblia in tropical regions of Iran and neighboring countries.

Livestock are commonly affected by gastrointestinal protozoan parasites, including Giardia duodenalis. In small ruminants, G. duodenalis infection may decrease carcass weight and dressing percentage. Current study was done to determine the prevalence, assemblage distribution, and zoonotic significance of Giardia infection in sheep and goats in Shiraz, southwestern Iran. In total, 200 fecal samples were randomly collected from 100 sheep and 100 goats in 10 farms in Shiraz, southwest of Iran (June 2021-February 2022) and directly examined for G. duodenalis cyst by saline/iodine wet mount examination using a light microscope (400 ×). Positive samples were further genotyped using a nested-PCR and sequencing methods. A mean prevalence of 5.5% (11/200) was estimated for G. duodenalis infection among small ruminants, with 7% and 4% for sheep and goats, respectively. Next, 9 out of 11 positive samples were amplified and only 5 were successfully sequenced at the tpi locus. Our results showed that 80% (4/5) isolates belonged to assemblage E, while only 20% (1/5) were associated with the assemblage A (subtype AI). Of note, 2 E assemblages were isolated from goats and 2 E assemblages plus 1 assemblage A were isolated from sheep. The major finding in the present study was the isolation of assemblage A from sheep in Shiraz, highlighting the zoonotic transmission of Giardia infection in the study area. In general, the information mentioned in the present study is very limited and more extensive studies in this field are needed to achieve more conclusive results.

Giardia duodenalis is a well-known flagellated parasite and the causative agent of protozoal diarrhea in animals and humans worldwide. Current study was aimed at determination of G. duodenalis prevalence, genetic variation and zoonotic significance in various species of rodents in Shiraz, southwestern Iran. In brief, 120 fecal specimens were collected from rodents (Rattus rattus, Rattus norvegicus, and Mus musculus) during May up to November 2021 and microscopically examined for Giardia cysts. Further molecular characterization of positive samples was done by nested-PCR, followed by nucleotide sequencing of the triose phosphate isomerase (tpi) gene. A total prevalence of 3.3% (4/120) was observed in rodents, with highest rate in black rats [5% (2/40)]. Regarding brown rats and house mice, only one sample was found to be positive, showing 2.5% and 2.5% prevalence, respectively. It is noteworthy that Giardia B and G assemblages were found in black rats (one case/genotype), whereas the only positive samples from brown rats and house mice were characterized as assemblage G. The major findings of the present study were the presence of both zoonotic and non-zoonotic Giardia assemblages in examined rats in Shiraz and the potential of black rats to harbor Giardia infection to humans. These concerns should be taken seriously in terms of public health. Nevertheless, the true epidemiology and assemblage distribution of Giardia is still open to question.

Background and Objectives: Nowadays, high-level aminoglycosides and ampicillin resistant Enterococcus species are among the most common causes of nosocomial infections. The present study was conducted to determine the prevalence of high-level resistance to aminoglycosides and ampicillin among clinical isolates of Enterococcus species in Ardabil, Iran. Materials and Methods: In this cross–sectional study, a total of 111 Enterococcus species were collected from different clinical specimens between 2013 and 2015. Enterococcus species were identified using standard phenotypic and genotypic methods. BHI agar screen and agar dilution methods were used for detection of high-level gentamicin and streptomycin resistance (HLGR and HLSR) and minimal inhibitory concentration (MIC) of ampicillin, respectively. Results: Of 111 clinical isolates, 59 (53.2%) and 25 (22.5%) isolates were E. faecalis and E. faecium, respectively, based on the PCR results. Totally, 60.3% and 56.7% of isolates were HLGR and HLSR, respectively, as well as 51.35% were HLGR plus HLSR. Among HLGR isolates, 36 (61.01%), 18 (72%) and 13 (48.14%) were E. faecium, E. faecalis and non-faecalis non-faecium species, respectively. Among HLSR isolates, 33 (55.93%), 16 (64%) and 14 (51.85%) were E. faecalis, E. faecium and non-faecalis non-faecium species, respectively. All HLGR isolates contained aac(6′)Ie-aph(2″)Ia gene. Overall, the prevalence of high-level ampicillin resistance among Enterococcus species was 17.1%. For E. faecalis, E. faecium and non-faecalis non-faecium species, ampicillin resistance rates were as follows: 11 (40.74%), 7 (28%) and 1 (1.69%), respectively. For aminoglycoside antibiotics, the resistance rate was significantly higher in E. faecium isolates and for ampicillin it was higher in E. faecalis isolates. Conclusion: The frequency of high-level aminoglycoside resistant enterococcal isolates in our hospital was high and significant ampicillin resistance was noticed. This would require routine testing of enterococcal isolates for HLAR and ampicillin susceptibility.

Background and Objectives: Nowadays, high-level aminoglycosides and ampicillin resistant Enterococcus species are among the most common causes of nosocomial infections. The present study was conducted to determine the prevalence of high-level resistance to aminoglycosides and ampicillin among clinical isolates of Enterococcus species in Ardabil, Iran. Materials and Methods: In this cross–sectional study, a total of 111 Enterococcus species were collected from different clinical specimens between 2013 and 2015. Enterococcus species were identified using standard phenotypic and genotypic methods. BHI agar screen and agar dilution methods were used for detection of high-level gentamicin and streptomycin resistance (HLGR and HLSR) and minimal inhibitory concentration (MIC) of ampicillin, respectively. Results: Of 111 clinical isolates, 59 (53.2%) and 25 (22.5%) isolates were E. faecalis and E. faecium, respectively, based on the PCR results. Totally, 60.3% and 56.7% of isolates were HLGR and HLSR, respectively, as well as 51.35% were HLGR plus HLSR. Among HLGR isolates, 36 (61.01%), 18 (72%) and 13 (48.14%) were E. faecium, E. faecalis and non-faecalis non-faecium species, respectively. Among HLSR isolates, 33 (55.93%), 16 (64%) and 14 (51.85%) were E. faecalis, E. faecium and non-faecalis non-faecium species, respectively. All HLGR isolates contained aac(6´)Ie-aph(2″)Ia gene. Overall, the prevalence of high-level ampicillin resistance among Enterococcus species was 17.1%. For E. faecalis, E. faecium and non-faecalis non-faecium species, ampicillin resistance rates were as follows: 11 (40.74%), 7 (28%) and 1 (1.69%), respectively. For aminoglycoside antibiotics, the resistance rate was significantly higher in E. faecium isolates and for ampicillin it was higher in E. faecalis isolates. Conclusion: The frequency of high-level aminoglycoside resistant enterococcal isolates in our hospital was high and significant ampicillin resistance was noticed. This would require routine testing of enterococcal isolates for HLAR and ampicillin susceptibility.

This is the first study aiming to determine the therapeutic effects of the Sambucus ebulus aquatic extract as an antileishmanial herbal drug and evaluate the immune responses in Leishmania major major infected BALB/c mice. The antileishmanial activity of S ebulus aquatic extract was evaluated using MTT test as well as parasite rescue and transformation assay. Footpad swelling and parasite load of infected mice were measured by several techniques. The immune responses were evaluated by measuring the levels of IFN-γ, IL-4, nitric oxide and arginase. The results indicated that S. ebulus can significantly decrease L. major promastigotes and amastigotes viability, but it was not toxic to macrophages. The lesion size, parasite burden and the level of ARG decreased in the treated infected mice, while the IFN-γ-to-IL-4 ratio and the level of NO increased significantly. Altogether, the S. ebulus extract is an effective compound for killing Leishmania parasite without excessive toxicity to the host cells and can cure the CL by switching the host immune responses towards Th1 response. Thus, it may be a perfect therapeutic option for CL treatment.

Schwann cells (SCs), the supporting cells of the peripheral nerves, are indispensable for regenerating the peripheral and central nervous system. Copious preparation of these cells in a well-defined manner is to be a privileged position. SCs cultivation is overwhelmed by contaminating fibroblasts which are often outgrowing as the predominant cell type in an in vitro culture. This study introduces a technically simple and efficient procedure for SCs isolation and enrichment based on implementing recombinant and defined supplements. Collected adult rat sciatic nerves were cultured for 10 days as in vitro predegeneration. After dissociation and plating, the medium changed to knockout serum replacement supplemented DMDM/F12 medium containing various growth factors. The whole procedure took 3 weeks and SCs purity was then evaluated through implementing specific cytoplasmic and membranous markers. The viability of enriched SCs were evaluated by MTT assay. Within 10 days, over 99 % homogenous SCs were achieved and confirmed through immunofluorescence staining and flow-cytometry for P75(NTR) and S100 markers, respectively. MTT data revealed that the viability and metabolic activities of purified SCs were increased in expansion medium. This study provides a technically easy and efficient method with the benefits of not utilizing bovine serum or other animal products for SCs isolation and enrichment.

Background & objective: Pneumocystis pneumonia (PCP) is responsible for pulmonary infection in immunocompromised patients. This study aimed to investigating the frequency of Pneumocystis colonization in patients hospitalized in the intensive care unit (ICU) and evaluating the relationship between PCP and Pneumocystis coloniza­tion. Methods: In the current cross sectional study bronchoalveolar lavage (BAL)fluids of 100 patients were collected from surgery and neurosurgery ICUs with different underlying corticosteroid therapy conditions. Patients were divided into 2 groups (patients who received corticosteroids and not received corticosteroids). Direct examination on BAL fluids was performed by the Gomori methenamine silver andGiemsa staining techniques. Additionally, 2 filtered air samples of the 2 above mentioned units were collected. A nested-PCR targeted mtLSUrRNA gene and sequencing were used to identify Pneumocystis spp. Results: In direct microscopy, 31 out of 100 hospitalized patients (31%) showed positive results. Twenty-three (46%) of smear positive patients were from the group of patients who received corticosteroid, the other 8(16%) were from the group of patients who didn’t receive corticosteroids (P= 0.001). Pneumocystis jirovecii DNA was detected in 77 out of 100 BAL samples by nested-PCR (77%) in which 40(52%) and 37(48%) samples were obtained from the patients who received and not received corticosteroids, respectively. Pneumocystis genome was found in 1 of the 2 filtered air samples. Conclusion: A significant number of patients who received corticosteroids were also colonized by P. jirovecii that may predispose to PCP or be transmitted to susceptible patients. A significant relationship was observed between the mean hospital stay and detection rate.

Animal and human fascioliasis is a health and economic problem in few of tropical and subtropical areas of the world, including Iran. The present study aimed to determine the genotype diversity of Fasciola isolates in different hosts from Gilan province, northern Iran, and compare it with those isolates from southwestern Iran. Forty-eight adult Fasciola spp. were collected from cattle, sheep, and goats from slaughterhouse in Talesh, north of Iran. DNA was extracted from each fluke and PCR-RFLP was used to find out the species of the isolates. The ribosomal ITS1 and ITS2, and mitochondrial genes of NDI and COI from individual Fasciola isolates of each host were PCR-amplified and the PCR products were sequenced. Genetic variation within and between the isolates was evaluated by comparing the sequences of ribosomal and mitochondrial genes. For analysis of phylogenetic diversity of the flukes, phylogenetic trees were constructed, using ITS1, ITS2, NDI, and COI sequences of the isolates. Based on PCR-RFLP profile, 5 (22.7%) of the total of sheep isolates and 18 (90%) of cattle isolates were identified as F. gigantica and other remaining samples from sheep, cattle and goats were identified as F. hepatica. Based on ITS1 and ITS2 sequences, six and seven nucleotide polymorphism were respectively noted in the isolates. On the other hand, CO1 region sequences showed considerable variation, which laid Talesh (north) isolates in a separate cluster. Findings of the study showed that the sequences of CO1 isolates from north and southwest have substantial differences mainly in CO1 region.

Background: Garlic and Zataria multiflora boiss (ZMB) are herbal medicines used traditionally in the treatment of infections, including candidal vaginitis. Objectives: This study aimed to examine the effect of garlic and ZMB vaginal cream 2%, in the treatment and recurrence of candidal vaginitis. Methods: This randomized, double-blinded clinical trial was conducted on 240 married women with candidal vaginitis. The patients used 5 gr garlic, ZMB or clotrimazole vaginal cream 2% daily, for 7 days. Complaints of the patients, clinical observations and laboratory parameters were recorded before treatment, on days 7 and 30, after treatment. The data were analyzed by SPSS v. 21 through chi-square, Fisher, ANOVA with repeated measures, McNemar and Kruskal-Wallis tests. A P 0.05). Mean scores of candidal symptoms reduced significantly in all groups at on 7 and 30, after treatment (P < 0.001). Conclusions: Garlic and ZMB vaginal cream 2% can be used as an effective treatment option in cases of drug resistance and, also, by the people who are interested in treating with herbal medicines.

Methicillin-resistant Staphylococcus aureus (MRSA) is a major cause of health care-associated infections. In this study, a total of 173 nurses was screened for S aureus nasal colonization, of which 8 (4.6%) were MRSA carriers. Among the MRSA isolates, 6 were mecA positive and oxacillin resistant, and 2 were mecA-positive, oxacillin-susceptible (OS-MRSA) strains. Reports of the OS-MRSA strains are increasing worldwide. To the best of our knowledge, this study is the first report on the occurrence of OS-MRSA strains in Iran.

Objective(s): Predegeneration is a standard technique to obtain mitotically activated and enriched cultures of Schwann cells (SCs). This study, for the first time, evaluated the impact of various duration of predegeneration on cell yield and enrichment of SCs from dog peripheral nerve. Materials and Methods: Dog sural nerves were subjected to 5, 10, 15 day-long in vitro predegeneration. The total cell yield and the purity of SCs were evaluated in each group on the first and seventh day after plating. Results: The maximum and minimum numbers of cells were counted in 15 day-long predegene-ration and control groups which underwent no predegeneration. The 10 day-long in vitro predegeneration group with 80±0.5% SCs enrichment had the best purity after plating day and could maintain its purity with elapsing on cultures. Conclusion: 10 day-long predegeneration results in the higher cell number and the better and prolonged purity of SCs in culture.

Visceral leishmaniasis (VL) is caused by Leishmania infantum in Mediterranean basin and is an endemic disease in some parts of Iran. Canines are the main reservoirs of VL in most of the endemic areas. Different serological methods have been introduced for diagnosis of canine visceral leishmaniasis (CVL).In this survey a Fucose-Mannose Ligand (FML) ELISA, using native L. infantum antigen, was developed and its validity for detection of infected dogs in comparison with direct agglutination test (DAT) and PCR was evaluated.Blood samples of sixty ownership dogs (≤ 3 years old) were collected from Meshkin-shahr district in Ardabil province, North-west of Iran. Sera were separated for serological assays (DAT and FML-ELISA) and the buffy coats were collected for molecular evaluation.Two out of the 60 (3.33%) samples were found to be positive (antibody titer of ≥ 1/320) in DAT while seven of the 60 (11.66%) samples were positive by FML-ELISA. Nine out of 60 (15%) buffy coat samples showed a band about 680 bp indicative of L. infantum in PCR. Three out of 60 dogs had Kala-azar symptoms and were positive by PCR and FML-ELISA, while two of these three dogs had antibody titers1/320 in their serum samples. The sensitivity and specificity of FML-ELISA for the detection of CVL in both symptomatic and asymptomatic dogs were found to be 77.8% and 100%, respectively.Considering the acceptable sensitivity and high specificity of FML-ELISA, use of this serological method can be recommended for epidemiological surveys of CVL.

Fascioliasis is a zoonose parasitic disease caused by Fasciola hepatica and Fasciola gigantica and is widespread in most regions of the world. Ectopic fascioliasis usually caused by juvenile Fasciola spp., but in recent years a few cases of tissue-embedded ova have been reported from different endemic areas. A 79-year-old Iranian man resident in Eird-e-Mousa village from Ardabil Province, north-west of Iran, complained with abdominal pain, nausea, and intestinal obstruction symptoms referred to Ardabil Fatemi hospital. In laparotomy multiple intestinal masses with peritoneal seeding resembling of a malignant lesion were seen. After appendectomy and peritoneal mass biopsy with numerous intraperitoneal adenopathy, paraffin embedded blocks were prepared from each tissues. A blood sample was taken from the patient 5 months later for serological diagnosis. Histopathological examination of sections showed fibrofatty stroma with dense mixed inflammatory cells infiltration and fibrosis in peritoneal masses. Large numbers of ova of Fasciola spp. were noted with typical circumscribed granulomas. Despite of anti-fasciola treatment, IHA test for detecting anti F. hepatica antibodies was positive 5 months after surgery with a titer of 1/128. Due to multiple clinical manifestation of extra-hepatic fascioliasis, its differential diagnosis from intraperitoneal tumors or other similar diseases should be considered.