behnam mohammadi-ghalehbin

Due to the increasing cohabitation of humans with infected animals, such as pets or wildlife, human infection with helminthic parasites has become an important emerging health concern. This study aimed to investigate the prevalence of helminthic parasite infections in canines in the Mughan Plain area of Ardabil Province, Iran. A total of 87 samples were collected from different canines, including stray dogs, foxes, and jackals, in the Mughan Plain area of Ardabil Province, Iran. The organs of the animals were sectioned and examined macroscopically for helminth parasitic infections. Isolated helminth species were morphologically identified using a valid identification key. The results showed that that Mesocestoides sp. was the most prevalent helminth species, with a frequency of 30 in canids (34.5%), including 8 red foxes (Vulpes vulpes, 53.33%), 12 jackals (Canis aureus, 48%), and 10 stray dogs (Canis familiaris, 21.3%). In addition, a high frequency of Toxascaris leonina was observed in 13 canines (14.9%), including 2 foxes (13%), 4 jackals (16%), and 7 wild dogs (14.9%). The prevalence of Echinococcus granulosus was reported in 9 canines (10.3%), including 1 jackal (4%) and 8 wild dogs (17.02%). The results of this study indicate that Mesocestoides sp. is the most common helminth species in canids. Our findings indicate that canines in Iran serve as both intermediate and final definitive hosts for several harmful parasites, posing a risk to humans and animals health. Further research and interventions are needed to better understand the transmission dynamics and to develop effective strategies for helminth control in canids.

Here we report biliary fascioliasis in two women 27 and 54 years old from Ardabil Province, Iran who presented with nausea, anorexia, weight loss, and pain between shoulder blades. Endoscopic ultrasound showed distal common bile duct strictures (CBD) and a lesion in its proximal. Endoscopic retrograde cholangiopancreatography (ERCP) was performed and live parasites were diagnosed and successfully managed into the duodenal lumen. The clinical findings of the patient improved after the procedure. This report emphasizes that the prevalence of Fasciola in Ardabil is predictable and this infection could be simultaneously diagnosed and treated by the ERCP.

 The available drugs for the treatment of leishmaniasis are highly toxic and extremely expensive, with low efficiency; therefore, the development of effective therapeutic compounds is essential.

 The present study aimed to explore the antileishmanial effects of ethyl acetate extract, methanol extract, and fractions 1-4 (F1-F4) of Ferula tabasensis, alone or in combination with shark cartilage extract (ShCE), on L. major in vitro.

 In this study, ethyl acetate, methanol, and n-hexane extracts were extracted from the aerial roots of F. tabasensis by the maceration method. The silica gel column chromatography was used to separate n-hexane extracts at varying polarities (F1-F4 fractions). Subsequently, the effects of extracts and fractions against promastigotes were assessed by the parasite counting method microscopic inhibition test and MTT assay. Besides, their effects on the infected macrophage cells and the number of amastigotes were investigated. Cytotoxicity was evaluated in non-infected J774A.1 macrophage cells. Finally, apoptosis induction of promastigotes, including infected and non-infected macrophages, was evaluated.

 The results indicated the highly potent activity of F. tabasensis extracts and F1-F4 fractions, alone or in combination with ShCE, against L. major promastigotes and amastigotes in a dose-dependent manner (P < 0.05). The F1 fraction and methanol extract showed markedly higher toxicity compared to the other extracts and fractions, with 50% inhibitory concentrations (IC50/72h) of 2.4 ± 0.29 and 2.9 ± 0.55 µg/mL against promastigotes and 1.79 ± 0.27 µg/mL and 1.39 ± 0.27 µg/mL against amastigotes (P < 0.001). Moreover, they had a high selectivity index (SI) due to the low toxicity of macrophages (P < 0.0001). The results of flow cytometry indicated that the percentages of apoptotic promastigote cells in contact with IC50 concentrations of F1 and methanol extract alone after 72 h were 43.83 and 43.93%, as well as 78.4%, and 65.45% for their combination with ShCE, respectively.
Also, apoptosis of infected macrophages induced by F1 and methanol extracts was estimated at 68.5% and 83.7%, respectively.

 In this study, the F1 fraction and methanol extract of F. tabasensis showed potent efficacy against L. major, associated with low toxicity and apoptosis induction. Therefore, they can be promising therapeutic candidates in future animal and even human studies.

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.

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.

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.

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.

Toxocariasis is a worldwide-distributed helminth parasitic infection. This study aimed to evaluate the seroprevalence of human toxocariasis in children living in Ardabil Province, North-West of Iran.

A seroepidemiological study to evaluate human toxocariasis among urban and rural populations of Ardabil County, North-West of Iran using ELISA test was carried out from 2019 to 2020. The study population was 472 children (185 females and 287 males) aged between 5 and 15 yr old. Immunoglobulin G antibodies against Toxocara spp. were analyzed by ELISA test.

Of the 472 collected serum samples, 66 (14.0%) were positive for antibodies against human toxocariasis. The highest prevalence of infection was observed in children with age groups of 5-6 years. Data demonstrate higher seroprevalence among males (15.3%) than females (11.8%). Rural areas prevalence was significantly higher (24.4%) than the urban area (8.65%). The rate of the diseases was 22.8% in children having history of contact with dog.

Toxocariasis is prevalent in the children of Ardabil region. The present study can increase the awareness of the population about the risk of zoonotic diseases as well as the anthelmintic treatment of dogs by veterinarians and, more importantly, the control of stray dogs.

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.

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.

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.

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.

 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.   

 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.

 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. 

 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.

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.

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.

Results of the study showed that compound A10, 4-(3-chlorophenyl)-6-methyl-N-phenyl2-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).

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.

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.

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.

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.

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.

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.
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.
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.
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.

Canine visceral leishmaniasis (CVL) caused by Leishmania infantum is endemic in the northwest and south of Iran. An appropriate vaccine can help to prevent and control visceral leishmaniasis in both humans and animals. Few studies have confirmed that the fucose-mannose ligand (FML) antigen of Leishmania donovani produced protective immunity in dogs against CVL.

To evaluate the immune responses of vaccinated dogs against FML antigen of L. infantum.

We isolated the FML antigen from native L. infantum and vaccinated the dogs with FML-saponin in an endemic area of VL in Iran to evaluate the immune responses of vaccinated dogs against this antigen.

Our results indicated a significant increase in the expression of IFN-γ, IL-10 and IL-13, but not IL-12A, gene transcripts in PBMCs of FML-saponin vaccinated dogs in comparison with controls. Our findings showed a significant difference in the ratio of IFN-γ/IL-10 mRNA expression in FML-saponin vaccinated dogs in comparison with two control groups. Moreover, a significant level of anti-FML antibodies was detected in serum of vaccinated dogs.

These findings showed that FML-saponin stimulates both Th1 and Th2 immune responses with predominant Th1 and strong humoral immune responses to produce protective immunity against CVL.

Despite continued and comprehensive planning of the world health organization (WHO), intestinal parasitic infections are a serious public problem in developing countries. Due to the high prevalence of cancers in Ardabil province and subsequently the high possibility of intestinal parasitic infections among the people, the aim of this study was to assess the frequency of intestinal parasites in patients with malignancy in this area.
In this cross-sectional study, 100 fecal samples were collected from patients with cancer during February to September 2015. The specimens were examined for intestinal parasites using direct smear, formol-ethyl acetate concentration, agar plate culture and Ziel-Neelsen staining technique.
The overall frequency of intestinal parasitic infections in studied cancer patients was 10%. The infection rates of detected intestinal parasites were Cryptosporidium spp. oocyst 4%, Blastocystis hominis 3%, Giardia lamblia 2% and Taenia spp. 1%.
Despite the low frequency of intestinal parasites, there is a need to screen cancer patients for some important parasitic infections such as Cryptosporidium spp. and Strongiloides stercoralis because of irreparable effects of those parasites on thepatients and to increase awareness among clinicians regarding the occurrence of parasitic infections in these patients.

Human toxocariasis is contained in the list of neglected diseases. The infection occurs after ingestion of embryonated eggs in contaminated soil. The present study was carried out to estimate the extent of soil contamination with Toxocara spp. eggs in the public places.
Soil samples were collected randomly from 41 public places in various parts in and around of Ardabil, Iran, between March 2013 and March 2014. Data were examined by microscopy following sodium nitrate flotation.
Of the 200 collected soil samples, 35 (17.5%) were positive for soil parasites. The eggs of Toxocara spp. were found in 14 (7%) soil samples.
This investigation gives baseline knowledge regarding soil contamination with Toxocara spp. eggs in Ardabil city and provides information for local control of toxocariasis.

Visceral leishmaniasis (VL) or kala-azar is a protozoan disease caused by some species of Leishmania donovani complex. Mediterranean type of the disease is endemic in various parts of Iran. A cross-sectional study was designed to determine the seroprevalence of VL among asymptomatic adult population in Meshkin-Shahr area from the Northwest of Iran as an endemic focus of VL.
Methods & Materials: Altogether, 180 blood samples were collected from asymptomatic adults’ population throughout 2015. Before sampling, a questionnaire was separately completed for each individual. All the collected blood samples were examined by direct agglutination test (DAT) after plasma separation. Anti-Leishmania infantum antibodies at titers 1:100 to 1:1600 was considered as L. infantum infection, while the cut-off titer of ≥ 1:3200 with specific signs and symptoms was considered as VL.
From 180 collected plasma samples, nine (5%) of them showed anti-Leishmania antibodies at titers 1:400 and higher. Distribution of anti-Leishmania antibodies titers was 1:400 (n = 2), 1:800 (n = 4), and 1:1600 (n = 3). All of the seropositive cases were observed among females. All the seropositive individuals had not a history of kala-azar. The highest seropositivity rate was observed among the age group of 13-23 years old. No changes in titers of anti-Leishmania antibodies observed after collected the seropositive blood samples again and tested by DAT with 1-month interval.
Visceral Leishmania infection is relatively high among adult people reside in Meshkin-Shahr area without any clinical manifestations. Asymptomatic VL infection is very important in immunocompromized individuals such as HIV-positive cases; these patients are at risk to manifesting clinical signs and symptoms of VL. Therefore continuing serological surveillance for detection of visceral Leishmania infection should be recommended in the endemic foci of VL.

There are many genera of free-living amoeba in the environment, but members of only four genera (Naegleria, Acanthamoeba, Balamuthia and Sappinia) have an association with human infection. Water, soil and air are main sources of infective types of these pathogenic organisms for human.

Totally, 30 samples were collected from the surface water sources of Shiraz city, the capital of Fars province, during July and August 2009. The samples were filtered and their sediments were cultured on non-nutrient agar medium and seeded with non-pathogen Escherichia coli. Then, they were incubated at three different temperatures, 22˚C, 37˚C, and 44˚C. The media were checked with invert microscopy and amoebae were recognized by phase–contrast microscopy and observed by light microscopy after Trichrome staining. Polymerase chain reaction (PCR) was performed for molecular detection.

Of the 30 samples, 29 were recognized morphologically as Acanthamoeba, the characteristics of 20 of which were confirmed by PCR. The growth rate of amoeba in 22˚C was more than 37˚C. Eight of the samples grew at 44˚C, but flagellate forming test and PCR were negative for Naegleria fowleri. Two of them were identified morphologically as Balamuthia and Sappinia.

Since Fars province is located in the subtropical region where there are a lot of parks and green areas with surface water, the potential risk of diseases caused by free-living amoebae should be considered. Further investigations about various aspects of these important opportunistic protozoa are recommended especially for establishment of appropriate prevention tools.

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. 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. 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. 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 inMediterranean basin and is an endemic disease in some parts of Iran. Canines are the mainreservoirs of VL in most of the endemic areas. Different serological methods have beenintroduced for diagnosis of canine visceral leishmaniasis (CVL). In this survey aFucose-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 titers >1/320 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.

Cryptosporidium is an intracellular – extracytoplasmic parasite that has taken much attention in last 20 years as a clinically important human pathogen. Cryptosporidial infection can be transmitted from fecally contaminated food or water and from animal-human or human-human contact. In immunocompromised persons, the illness is much more severe such as debilitation, fatigue, cholera-like diarrhea, severe abdominal cramps, low-grade fever, severe weight loss and Anorexia. Because there was no regional study about cryptosporidiosis in Ardabil, we carried out this survey to determine the prevalence of cryptosporidiosis among the children hospitalized in Ardabil. This descriptive and analytical study was carried out on 371 patients in Sabalab and Aliasghar hospitals of Ardabil between 2004 and 2005. A questionnaire was filled for each patient. Stool samples were examined by concentrated formal - ether method and stained with modified Ziehl-Neelson method. The data were analyzed with SPSS (ver 11) using Chi-square test. We analyzed 371 stool samples from children with diarrhea. Cryptosporidium oocysts were detected microscopically in 15 samples. Its prevalence was 4.04% in infected patients. 66.7% of the infected ones were at the age of 6 to 24 months, 20% 25-48 months, and 13.3% 49-72 months. Condusion: Because cryptosporidiosis was more prevalent at the age of 6-24 months, health education is more necessary for their mothers.