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Background and Objective: Cutaneous leishmaniases with two forms of rural and urban is the endemic diseases and as a health problem in our country. Identification of parasite species and type of disease is very important for treatment of disease as well as for planning of control program. The microscopic observations by Giemsa-stained smears is the most common laboratory test for the diagnosis of cutaneous leishmaniasis, but the determination of parasite species is impossible and utilization of other ways such as biochemical and molecular methods is required. This study was carried out to determine the parasite species caused cutaneous Leishmaniasis by Nested PCR in Damghan, Iran. Materials and Methods: This descriptive study was performed on 67 patients with dermal lesions that referred to Damghan health center laboratory in Iran during 2008. The patient's information were recorded in questionnaire. DNA of Giemsa-stained slides from patients was extracted and evaluated by specific primers of kinetoplast DNA using Nested PCR. Results: Leishmania parasites were observed in 57 patients under light microscope. The 10 patients were infected by other dermal diseases. The PCR result showed the parasite presence in lesions of 57 patients is Leismania major. 54% of patients were male and 46% were female. 72% of the patients were lived in rural areas. 50.9% of disease was observed in over 25 years old patients. Hands were the most common region of ulcer (44.7%). 48% of the patients had one ulcer and the other patients had two or more ulcers. High prevalence (31.6%) of disease was observed in October. Conclusion: This study showed that zoonotic cutaneous leishmaniasis to be prevalent in this area and Nested PCR method is a sensitive and accurate to leishmania species characterization.

In this work, pomegranate juice was adulterated manually by adding sour cherry and red grape juices. Physicochemical properties (brix, acidity, formalin index, reducing sugar, and total sugar) and Volatile Organic Compounds (VOCs) gas chromatography profile (extracted by PPy-Ag nano-fiber) of pure pomegranate juice and adulterated pomegranate juice were evaluated and used to find adulteration. PPy-Ag nano-fiber as a Head-Space Solid Phase Micro Extraction-Gas Chromatography (HS-SPME-GC) method was used to extract and detect the VOCs profile (total peak area and total peak height) of different juices. The Box-Behnken Design (BBD) was used to study the effects of Pomegranate Juice (PJ), Sour Cherry Juice (SCJ) and Grape Juice (GJ) percent (V/V%) in the adulterated mix juice physicochemical properties and GC-profile. Results showed that the percent of pomegranate juice, sour cherry juice, and grape juice affect the juice physicochemical property, and there are relations between the physicochemical property of the juice and VOCs gas chromatography profile (total peak area and total peak height) that can help to find adulteration. The presented research makes it possible to detect adulteration of sour cherry and grape juice and their percent in the adulterated mix by analyzing the juice physicochemical properties and GC-profile. [ABSTRACT FROM AUTHOR]

The aim of this study was to evaluate the effects of inulin (IN) and resistant dextrin (RD) as fat replacer and prebiotic on gluten-free biscuit and its dough. To make the gluten-free biscuits, we used rice flour, corn flour and corn starch in the proportion of 3:1:1, respectively. The influence of prebiotics on the dough properties was studied via texture profile analysis including firmness, cohesiveness, adhesiveness, gumminess and springiness. Biscuit quality was assessed by spreading behavior, texture and surface characteristics, chemical properties, and sensory evaluation. Compared to the control, a significant increase in firmness (17.04 N to 52.85 N), cohesiveness (0.49 to 0.65) and gumminess (8.45 N to 32.71 N) of dough (except RD25) was observed when substitution percent of fibers increased. Adhesiveness and springiness did not have significant changes. Enhancing of fat replacement percentage caused significant changes compared to the control in hardness (9.60 to 24.52 N) and L* (58.79 to 56.94), a* (8.99 to 9.71), water activity (0.225 to 0.096), moisture (4.97% to 4.12%), total fat (12.65% to 3.90%), peroxide index (1.89 to 0.90 meq/kg), fiber (2.02% to 9.51%), carbohydrate (76.49% to 84. 63%), and calorie (443.38 to 396.52 Kcal). The consumers did not find significant differences in acceptability between the control biscuits and the biscuits with 25% of fat replaced by RD and IN except color and flavor that were better than the control. Gluten-free biscuits containing IN25 and RD25 were similar to the control biscuits, and they could have additional health benefits derived from IN and RD presence. [ABSTRACT FROM AUTHOR]

Background: Plant extracts and oils may act as alternatives to conventional pesticides for malaria vector control. The aim of this study was to evaluate the larvicidal activity of essential oils of three plants of Apiaceae family against Anopheles stephensi, the main malaria vector in Iran. Methods: Essential oils from Heracleum persicum, Foeniculum vulgare and Coriandrum sativum seeds were hydro distillated, then their larvicidal activity were evaluated against laboratory-reared larvae of An. stephensi according to standard method of WHO. After susceptibility test, results were analysis using Probit program. Results: Essential oils were separated from H. persicum, F. vulgare and C. sativum plants and their larvicidal activities were tested. Result of this study showed that F. vulgare oil was the most effective against An. stephensi with LC50 and LC90 values of 20.10 and 44.51 ppm, respectively. Conclusion: All three plants essential oil can serve as a natural larvicide against An. stephensi. F. vulgare oil exhibited more larvicidal properties. [ABSTRACT FROM AUTHOR]

Background: Appropriate methodology for storage biological materials, extraction of DNA, and proper DNA preservation is vital for studies involving genetic analysis of insects, bacteria, and reservoir hosts as well as for molecular diagnostics of pathogens carried by vectors and reservoirs. Here we tried to evaluate the utility of a simple filter paper-based for storage of insects, bacteria, rodent, and human DNAs using PCR assays. Methods: Total body or haemolymph of individual mosquitoes, sand flies or cockroaches squashed or placed on the paper respectively. Extracted DNA of five different bacteria species as well as blood specimens of human and great gerbil Rhombomys opimus was pipetted directly onto filter paper. The papers were stored in room temperature up to 12 months during 2009 until 2011. At monthly intervals, PCR was conducted using a 1-mm disk from the DNA impregnated filter paper as target DNA. PCR amplification was performed against different target genes of the organisms including the ITS2-rDNA of mosquitoes, mtDNA-COI of the sand flies and cockroaches, 16SrRNA gene of the bacteria, and the mtDNA-CytB of the vertebrates. Results: Successful PCR amplification was observed for all of the specimens regardless of the loci, taxon, or time of storage. The PCR amplification were ranged from 462 to 1500 bp and worked well for the specified target gene/s. Time of storage did not affect the amplification up to one year. Conclusion: The filter paper method is a simple and economical way to store, to preserve, and to distribute DNA samples for PCR analysis. [ABSTRACT FROM AUTHOR]

The problems caused by the pollution of the environment by petroleum polymers in recent years have caused researchers to think of replacing petroleum polymers with biodegradable and natural polymers. The aim of this research was to produce composite film of chitosan (Chit)/zero-valent iron (Fe) nanoparticles/oregano essence (Ess) (Chit/Fe/Ess). Central composite design was used to study physical, morphological, antioxidant and antimicrobial properties of films. The results showed that with the increase of iron nanoparticles and oregano essence, the thickness of the film increased. The moisture, solubility and water vapor permeability of the film decreased with the increase of iron nanoparticles and oregano essence. The results of the mechanical test showed that with the increase of iron nanoparticles and oregano essence, the tensile strength and elongation at break point decreased. Iron nanoparticles and oregano essence increased significantly the antioxidant activity of the film. The results of the antimicrobial activity of the prepared films show that the addition of iron nanoparticles and oregano essence enhanced the antimicrobial activity of the film against Escherichia coli and Staphylococcus aureus. X-ray diffraction analysis showed that iron nanoparticles were physically combined with chitosan polymer. Fourier transform infrared (FTIR) results confirmed the physical presence of iron nanoparticles and oregano essence in the polymer matrix. The results of scanning electron microscopy (SEM) showed that the surface of nanocomposite films is more heterogeneous than chitosan. Iron nanoparticles and oregano essence could delay the thermal decomposition of chitosan and increase the thermal stability of chitosan film., (© 2024 Wiley Periodicals LLC.)

In this study, barley sprout powder/pectin (BS/Pec) composite film was prepared. Quercetin (Qu) and vanadium oxide (V 2 O 5 ) nanoparticles were used to improve the physicochemical and structural characteristics of the film. The structural, physicochemical and thermal properties of the films were investigated by various techniques such as TGA, SEM, XRD, FTIR, texture analysis, etc. The thickness and tensile strength of the films increased from 120 μm to 2.4 MPa to 220 μm and 6 MPa respectively with the increase of V 2 O 5 nanoparticles and quercetin pigment. Nanoparticles of V 2 O 5 and quercetin decreased the moisture content of the film from 50% to 20%. Quercetin had little effect in reducing water vapor permeability (WVP), but V 2 O 5 nanoparticles had a significant effect in reducing WVP. The pure BS/Pec film had almost 30% antioxidant properties, which increased to 81% with the increase of quercetin. Adding quercetin and V 2 O 5 nanoparticles to the film increased the antimicrobial properties of the film against both Escherichia coli and Staphylococcus aureus bacteria. The SEM images showed the inhomogeneous surface of the BS/Pec film caused by BS powder fibers. The interactions between the components of the films (electrostatic type) was confirmed by FTIR results. The degradation temperature of the overall structure of the film in the presence of nanoparticles indicated the positive effect of nanoparticles in increasing the thermal resistance of the film. Investigating the crystal structure of the film showed that the BS/Pec film has an amorphous/crystalline or semi-crystalline structure. Considering that the prepared film has good mechanical properties and as well as antioxidant/antimicrobial properties, this film as an active composite can be used in food products packaging., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 Published by Elsevier Ltd.)

This paper investigates the addition of lecithin-emulsified black seed oil (BSO) nanoemulsions (LNEO) and whey protein isolate-stabilized Pickering emulsions (WPEO) to soy protein isolate (SPI)-based films and their effect on improving the shelf life of bread slices. The half-life of antioxidant activity, water vapor permeability, biodegradability, density, color difference, and film thickness significantly increased ( p  < .05) when BSO was added. However, the incorporation of BSO significantly reduced the solubility, tensile strength, strain to break (except for WPEO), and transparency ( p  < .05) of the samples. The interaction between SPI film and BSO-loaded nanocarriers, as well as the morphological properties of films, was evaluated using FT-IR and FE-SEM. SPI-based films containing LNEO-5% and WPEO-5% were selected based on their mechanical and barrier properties. The effect of films on the shelf life of bread slices was investigated for 17 days of storage. LNEO samples obtained the most acceptable results in the bread in terms of sensory evaluation and color properties. According to the results, bread slices packed in SPI film containing LNEO-5% showed no signs of mold growth until the 17th day of storage, whereas the sample packed in a low-density polyethylene bag began to corrupt on the 6th day. This study highlights the potential of BSO-loaded SPI films as a novel active packaging for the bakery industry., Competing Interests: The authors declare no competing interests., (© 2023 The Authors. Food Science & Nutrition published by Wiley Periodicals LLC.)

This research aimed to extract flaxseed mucilage (FM) and investigate its rheological properties (static and dynamic tests) compared with animal oil. In the next stage, the D-optimal design was applied to investigate the effect of FM (0-60%) and FF (0-30%) replacements with animal oil and cake flour, respectively on the sponge cake's physicochemical, textural, and sensory properties. According to the flow behavior test, FM showed Newtonian behavior while animal oil had pseudoplastic behavior. The results of the dynamic test showed with an increase in frequency, the loss modulus (G״) and storage modulus (G') of samples increased. However, G' was higher than G״ in all samples. By replacement of FM and FF, the moisture content, water activity, antioxidant capacity, crumb hardness, and cohesiveness of the samples increased while springiness, crust hardness, and specific volume decreased (P < 0.05). Lightness of samples with replacement of FF decreased (P < 0.05). The overall acceptance score was enhanced with an increase in FM substitution while it was decreased with the replacement of FF (P < 0.05). The amounts of fat, calories, and peroxide values were decreased in sponge cake with the incorporation of FF and FM (P < 0.05). In general, the substitution of FM (60%) and FF (28%) in the cake formulation as an optimized sample to make new products with low-calorie content is possible without significant decreases in product quality., (© 2023. The Author(s).)

This study aimed to investigate in situ exopolysaccharides (EPSs) production by Bacillus coagulans IBRC-M 10807 under different fermentation conditions to improve the technical-functional properties of whole wheat flour sourdough and obtain high-quality products. For this purpose, the effectiveness of four efficient factors including B. coagulans (8 Log CFU/g), FOS (0%, 2.5%, and 5% based on flour weight), fermentation temperature (30, 35, and 40°C), and fermentation time (12, 18, and 24 h) was investigated on the production of functional sourdough. Our work focused on optimizing probiotic sourdough by investigating probiotic viability, pH, total titratable acidity, antioxidant properties, and EPS measurement. The first optimal formulation for maximized production of the in situ EPSs by the numerical optimization included FOS 0%, B. coagulans IBRC-M 10807 8 Log CFU/g, fermentation temperature of 30°C, and fermentation time of 12 h. In this case, EPSs was 59.28 mg/g and probiotic was 10.99 Log CFU/g. The second optimal formula by considering the highest viability of probiotic together with EPS production was determined as FOS 4.71%, B. coagulans IBRC-M 10807, 8 Log CFU/g, fermentation temperature of 30°C, and fermentation time of 20 h. The predicted amount of the EPSs and probiotic viability via the second formulation were 54.4 mg/g and 11.18 Log CFU/g, respectively. Analyses of optimal sourdough using FTIR, SEM, and DSC revealed that FOS and probiotics significantly reduced the enthalpy of amylopectin retrogradation and delayed it compared to other samples. Therefore, improving the final product's technological capabilities and shelf life can be credited with potential benefits., Competing Interests: The authors certify that they have no conflict of interest concerning this manuscript., (© 2023 The Authors. Food Science & Nutrition published by Wiley Periodicals LLC.)

Amaranth selective hydrolyzed protein (ASPH) may improve sourdough properties and bread quality. In this regard, this study focused on investigating the influence of protein hydrolysates on sourdough fermentation and bread properties. Based on the findings, ASPH further increased Lactobacillus plantarum and Saccharomyces cerevisiae growth in sourdough compared with amaranth protein isolates and amaranth flour. ASPH at 5 g/kg resulted in sourdough with higher pH and total titratable acidity (TTA) after 20 h of fermentation at 30°C. The prepared sourdough using APH (S-ASPH) at 3 g/kg increased the specific volume (4.57 ml/g) and TTA (4.76 ml) while decreasing water activity, hardness, cohesiveness, and chewiness of the bread (S-ASPH-B) compared with the control. Moreover, transition temperature and enthalpy reduced whereas sensory properties and shelf life represented an increase with S-ASPH addition. Overall, the obtained data indicated the improvement of bread quality by S-ASPH sourdough., Competing Interests: The authors certify that they have no conflict of interests with respect to this manuscript., (© 2021 The Authors. Food Science & Nutrition published by Wiley Periodicals LLC.)

Forensically important flesh flies (Diptera: Sarcophagidae) often are not morphologically distinguishable, especially at the immature stage. In addition, female flies are quite similar in general morphology, making accurate identifications difficult. DNA-based technologies, particularly mitochondrial DNA (mtDNA), have been used for species-level identification. The cytochrome oxidase subunits I and II (COI-COII) sequences of Iranian Sarcophagidae are still unavailable in GenBank. In this study as many as 648 (540 males and 106 females) fly specimens from family Sarcophagidae, representing 10 sarcophagid species, including eight forensically important species were collected from seven locations in five Iranian provinces. Of these, 150 male specimens were identified based on both morphology of male genitalia and DNA sequencing analysis. Sequence data from the COI-COII regions for 10 flesh fly species collected in Iran were generated for the first time. Digestion of COI-COII region by restriction enzymes RsaI, EcoRV, and HinfI provided distinct restriction fragment length polymorphism profiles among the species and can serve as molecular markers for species determination. Phylogenetic analysis represented that the COI-COII sequences are helpful for delimitation of sarcophagid species and implementation in forensic entomology. However, the application of the COI-COII fragment as a species identifier requires great caution and additional species and markers should be studied to ensure accurate species identification in the future., (© The Author(s) 2019. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

In this study, biodegradable and edible films based on eggplant flour (EF) and corn starch (CS) were prepared using casting method at proportion of 0-100, 25-75, 50-50, 75-25 and 100-0%, w/w. The mechanical, barrier, physical, and biodegradability properties were evaluated. Tensile strength, elongation at break, Young's modulus, thickness, density and L ⁎ parameter of pure starch films were higher than those of other films. Solubility, water vapor permeability (WVP), moisture content and swelling index of films were augmented with the substitution of CS by EF. Color measurement of the edible films indicated that increasing the proportion of EF increased a ⁎ , b ⁎ and opacity values. The highest amount of water sorption was obtained for pure EF films. Moreover, the incorporation of EF accelerated films biodegradability compared to ones with only starch. In general, EF is a promising material for the formulation of edible and biodegradable films with adequate physical properties for food applications by direct contact., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Background: Enterobacter cloacae bacterium is a known symbiont of the most Anopheles gut microflora and nominated as a good candidate for paratransgenic control of malaria. However, the population dynamics of this bacterium within An. stephensi and its introduction methods to the mosquitoes have not yet been explored., Methods: Enterobacter cloacae subsp. dissolvens expressing green fluorescent protein and defensin (GFP-D) was used to study transstadial transmission and the course of time, larval habitat, sugar, and blood meal on dynamics of the bacterium in the mosquito life stages in the laboratory condition. The bacterial quantities were measured by plating samples and counting GFP expressing colonies on the Tet-BHI agar medium., Results: The E. cloacae population remained stable in sugar bait at least for eleven days whereas it was lowered in the insectary larval habitat where the bacteria inadequately recycled. The bacterium was weakly transmitted transstadially from larval to adult stage. The bacterial populations increased smoothly and then dramatically in the guts of An. stephensi following sugar and blood meal respectively followed by a gradual reduction over the time., Conclusion: Enterobacter cloacae was highly stable in sugar bait and increased tremendously in the gut of female adult An. stephensi within 24h post blood meal. Sugar bait stations can be used for introduction of the transgenic bacteria in a paratransgenic approach. It is recommended to evaluate the attraction of sugar bait in combination with attractive kairomones as well as its stability and survival rate in the semi-field or field conditions.

Members of the Culex (Culex) pipiens assemblage are known vectors of deadly encephalitides, periodic filariasis, and West Nile virus throughout the world. However, members of this assemblage are morphologically indistinguishable or hard to distinguish and play distinct roles in transmission of the diseases. The current study aimed to provide further evidence on utility of the two most popular nuclear (ITS2-rDNA) and mitochondrial (COI barcode region) genetic markers to identify members of the assemblage. Culex pipiens assemblage specimens from different climate zones of Iran were collected and identified to species level based on morphological characteristics. Nucleotide sequences of the loci for the specimens plus available data in the GenBank were analyzed to find species specific genetic structures useful for diagnosis purposes. ITS2 region was highly divergent within species or populations suggesting lack of consistency as a reliable molecular marker. In contrast, sequence analysis of 710 bp of COI gene revealed three fixed haplotypes named here "C, T, H" within the assemblage which can be distinguished by HaeIII and AluI enzymes. There were a correlation between the haplotypes and the world climate regions, where the haplotypes H/T and C are present mainly in temperate and tropical regions of the world, respectively. In the New world, Australia, and Japan only haplotype H is found. In conjunction between tropical and temperate regions such Iran, China, and Turkey, a mix of C/H or C/H/T are present. Although, the haplotypes are not strictly species-specific, however, Cx. quinquefasciatus was mainly of haplotype C. Due to the lack of mating barrier and questionable taxonomic situation of the complex members, the mentioned haplotypes in combination with other morphological and molecular characters might be used to address the genetic structure of the studied populations., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Background: Linear dermatitis is endemic in Iran where most cases occur in the Caspian Sea coast and Fars province. The disease is caused by beetles of the genus Paederus which are active from early spring to beginning of autumn although its incidence rises from May to August. The classic taxonomy of Paederus spp. is based on the male genitalia that is very complex and needs expertise. In this study, we report a DNA-based method to discriminate Paederus fuscipes and Paederus littoralis (=syn: P. lenkoranus , P. ilsae )., Methods: Type specimens were collected from north and south of Iran. Molecular typing of the species was performed using restriction fragment length polymorphism (RFLP) analysis of polymerase chain reaction (PCR)-amplified fragments of mtDNA-COI., Results: Sequence analyses of the data obtained in this study showed significant DNA polymorphisms. There were 89 substitutions between COI sequences of the two species. The mtDNA-COI fragment comprises several useful species-specific restriction sites comprising Hae III that could result in distinctively different species-specific PCR-RFLP profiles. The Hae III enzyme cuts the 872 bp PCR amplicon of P. littoralis into 737 and 100 bp and two small nonvisible bands whereas it does not cut P. fuscipes amplicon into fragments., Conclusion: This study demonstrates that molecular typing is useful method and allows one to differentiate between two species and is recommended for discrimination of other Paederus species, which morphologically are indistinguishable or very difficult to be distinguished.

Background: Phlebotomus sergenti s.l. is considered the most likely vector of Leishmania tropica in Iran. Although two morphotypes- P. sergenti sergenti (A) and P. sergenti similis (B)-have been formally described, further morphological and a molecular analysis of mitochondrial cytochrome oxidase I (mtDNA-COI) gene revealed inconsistencies and suggests that the variation between the morphotypes is intraspecific and the morphotypes might be identical species., Methods: We examined the sequence of the ITS2-rDNA of Iranian specimens of P. sergenti s.l., comprising P. cf sergenti , P. cf similis , and intermediate morphotypes, together with available data in Genbank., Results: Sequence analysis showed 5.2% variation among P. sergenti s.l. morphotypes. Almost half of the variation was due to the number of an AT microsatellite repeats in the center of the spacer. Nine haplotypes were found in the species constructing three main lineages corresponding to the origin of the colonies located in southwest (SW), northeast (NE), and northwest-center-southeast (NCS). Lineages NCS and NE included both typical P. cf sergenti and P. cf similis and intermediate morphotypes., Conclusion: Phylogenetic sequence analysis revealed that, except for one Iranian sample, which was close to the European samples, other Iranian haplotypes were associated with the northeastern Mediterranean populations including Turkey, Cyprus, Syria, and Pakistan. Similar to the sequences of mtDNA COI gene, ITS2 sequences could not resolve P. sergenti from P. similis and did not support the possible existence of sibling species or subspecies within P. sergenti s.l. .

Recent epidemiological evidences revealed a higher rate of O blood group in the residents of malaria-endemic areas suggesting that groups A, B, and AB associated with a higher disease severity and fatality. Also recent data showed the low prevalence of AB group within the malaria-endemic residents in south of Iran and India. The aim of this study was to determine the ABO blood groups preference of Anopheles stephensi which is the main malaria vector in Iran, southwest Asia, and India. An. stephensi mosquitoes were fed either artificially on A/B/O/AB membrane blood feeders or directly on human volunteer hands and forearms of A/B/O/AB groups in a cage under lab conditions. Phenotype and genotype analyzes of 450-blood-fed mosquito specimens using agglutination and multiplex-allele-specific PCR revealed a significant blood preference of An. stephensi to AB group (40%) than other groups of A (24%), B (21%), and O (15%) in combination of both experiments. High preference of An. stephensi to AB group might increase malaria infection and fatality in this blood group and resulted in low frequency of AB group in the residents of malaria endemic areas. The data suggested that malaria vectors, like parasites may have selection pressure on human genotypes., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Recent epidemiological evidences revealed the higher prevalence of 'O' blood group in the residents of malaria-endemic areas. Also some data indicated preference of mosquitoes to 'O' group. The aim of this study was to determine ABO group ratio in the residents as well as ABO group preference of Anopheles in two malaria endemic areas in south of Iran. Agglutination method was used for ABO typing of residents. Field blood fed Anopheles specimens were tested against vertebrate DNA using mtDNA-cytB PCR-RFLP and then the human fed specimens were tested for ABO groups using multiplex allele-specific PCR. A total of 409 human blood samples were identified, of which 150(36.7%) were 'O' group followed by 113(27.6%), 109(26.7%), and 37(9.0%) of A, B, and AB groups respectively. Analyzing of 95 blood fed mosquitoes revealed that only four Anopheles stephensi had fed human blood with A(1), B(1), and AB(2) groups. Result of this study revealed high prevalence of O group in south of Iran. To our knowledge, it is the first ABO molecular typing of blood meal in mosquitoes; however, due to low number of human blood fed specimens, ABO host choice of the mosquitoes remains unknown. This study revealed that ABO blood preference of malaria vectors and other arthropod vectors deserves future research., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Background: Great gerbils, Rhombomys opimus, are the main reservoir host of zoonootic cutaneous leishmaniasis (ZCL) in Iran and neighboring countries. Based on morphological traits two subspecies R. opimus sodalis and R. opimus sargadensis have reported in the country. However, variation in infection rate and signs to Leishmania parasites, phenotype, size, and sexual polymorphisms demand more details to elucidate clearly the role of great gerbils in ZCL epidemiology., Methods: PCR-RFLP and PCR-direct sequencing were used to analyze mitochondrial DNA cytochrome B (mtDNA-cytB) gene structure of R. opimus collected from Golestan and Khorasan-e-Razavi Provinces in 2011 that are neighbor to Turkmenistan Country where ZCL is endemic in both sides of the borderline., Results: All of the specimens (n= 61) were morphologically or genetically similar to the typical R. opimus sodalis. However, there were 9 (1.5%) DNA substitutions throughout the 583 bp of the Cyt b gene of the samples sequenced comprising six DNA haplotypes. Maximum likelihood or neighbor joining phylogenetic trees inferred from the sequences could resolve the populations according to their subspecies as well as geographical origins., Discussion: The DNA polymorphisms in the great gerbils may correspond to the signs and infection rate in the animal. However, further studies are needed to match these six haplotypes with different signs and parasite sustaining following infection with L. major in the great gerbils.

Visceral leishmaniasis (VL) is an important health problem in Ardebil, where it borders Azerbaijan in the northwestern Iran. In spite of the presence of both cutaneous and visceral leishmaniasis (CL and VL) in northwestern Iran, previous researches have consistently revealed the etiologic agent of VL in the region to be Leishmania infantum. This is the first report of natural infection of Phlebotomus tobbi with L. infantum in Bilesavar district in the northern part of Ardebil province bordering Azerbaijan. Polymerase chain reaction (PCR) of kDNA, ITS1-rDNA, and CPB genes of the parasite followed by restriction fragment length polymorphism (RFLP) and gene sequencing analyses revealed presence of L. infantum in six out of 433 tested female sand fly specimens. Although sand flies of P. tobbi were infrequent, two out of 32 (6.25%) females captured in the area were found infected with the parasite. Phlebotomus perfiliewi transcaucasicus, the known vector of VL in the area, were the most dominant species but only four out of 273 (1.47%) tested were infected with L. infantum. This study showed that P. tobbi similar to P. perfiliewi transcaucasicus could play a significant role in the transmission of the L. infantum. However more investigations are needed to demonstrate that L. infantum is the only species circulating in the focus., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Accurate identification of animal reservoirs of transmissible diseases is an absolute requirement to any epidemiological survey of zoonoses and is essential for predicting species-specific population outbreaks and therefore to develop accurate ecological control strategies. The systematic status of the great gerbil (Rhombomys opimus) remains unclear, despite the fundamental role of these rodents as the main known reservoir hosts of the protozoan parasite Leishmania major in the epidemiology of zoonotic cutaneous leishmaniasis (ZCL) in central and south Asia. In the present work, we represent molecular evidence supporting the identification of at least two major lineages (subspecies) within the species of R. opimus in Iran. Analysis of the mitochondrial cytochrome b (cyt b) gene, revealed a range of 1-10% genetic variation among populations, which were well associated with biogeographic origins and subspecies. Results of laboratory cross hybridization between the subspecies and finding sympatric haplotypes of the two subspecies suggested that no pre- or post-zygotic barriers exist between the subspecies indicating that they still belong to a single taxon. However, the amount of genetic variations between populations/subspecies is high enough to lead them to speciation in future. Implications of such findings on the eco-epidemiology of ZCL in Iran are discussed., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Background: Using botanical insecticides as an alternative biocontrol technique for vector control is considered by some scientists., Materials and Methods: Chemical composition of the essential oil was analyzed using gas chromatography-mass spectrometry (GC-MS). In addition, the mosquito larvicidal activity of leaf essential oil of Cupressus arizonica was investigated against fourth instar larvae of laboratory-reared An. stephensi according to the method of the World Health Organization., Results: Of 46 constituents in the oil, limonene (14.44%), umbellulone (13.25%) and α-pinene (11%) were determined as the main constituents. Cupressus arizonica volatile oil showed significant larvicidal activity against An. stephensi with LC(50) and LC(90) values 79.30 ppm and 238.89 ppm respectively. Clear dose-response relationships were established with the highest dose of 160 ppm essential oil with almost 100% mortality., Discussion: The results from this study revealed that C. arizonica essential oil could be considered as a natural larvicide against An. stephensi. However, the field evaluation of the formulation is necessary.

Tick-borne relapsing fever (TBRF) is endemic in Africa and Eurasia and attributed to different Borrelia species. In the Central Asia and Middle Eastern countries, TBRF is caused mainly by Borrelia persica; however, other Borrelia species such B. microtti, B. latyschewii, B. baltazardi, and B. caucasica have also been described. The classic taxonomy of Borrelia spp. is based on the cospeciation concept that is very complex and rather confusing. In this study, we report two DNA-based methods to discriminate B. persica and B. microtti, the two main prevalent species in the region. Molecular typing of the species was performed using (i) restriction fragment length polymorphism analysis of polymerase chain reaction (PCR)-amplified fragments of either 16S-rRNA or glpQ genes, and (ii) species-specific PCR of glpQ gene. Sequence analyses of the data obtained in this study indicate that the glpQ gene is more variable than 16S-rRNA (6.9% vs. 1.2%); thus glpQ is a more useful marker for discrimination of B. persica from B. microtti. The 16S-rRNA fragment comprises only one useful species-specific restriction site (TaqI), whereas the glpQ fragment includes several species-specific restriction sites and its digestion by DraI, TaqI, EcoRV, HinfI, or SspI results in distinctively different PCR-restriction fragment length polymorphism patterns for the two species. Further, the species-specific primers amplified a 253-bp fragment for B. persica and a 451-bp one for B. microtti. Phylogenetic analysis of the data revealed that B. microtti and B. persica are associated to the African and new world RF agents, respectively. This study demonstrates that both typing methods are simple, sensitive, and fast, and that they allow one to differentiate between B. persica and B. microtti. This could prove that both methods are important and useful in monitoring of TBRF disease in endemic areas.

An epidemiological study was carried out on the vectors and reservoirs of cutaneous leishmaniasis in rural areas of Damghan district, Semnan province, central Iran, during 2008-2009. Totally, 6110 sand flies were collected using sticky papers and were subjected to molecular methods for detection of Leishmania parasite. Phlebotomus papatasi Scopoli was the common species in outdoor and indoor resting places. Polymerase chain reaction technique showed that 24 out of 218 P. papatasi (11%) and 4 out of 62 Phlebotomus caucasicus Marzinovskyi (6.5%) were positive for parasites Leishmania major Yakimoff and Schokhor. Twenty-one rodent reservoir hosts captured using Sherman traps were identified as Rhombomys opimus Lichtenstein (95%) and Meriones libycus Lichtenstein (5%). Microscopic investigation on blood smear of the animals for amastigote parasites revealed 8 (40%) rodents infected with R. opimus. L. major infection in these animals was then confirmed by polymerase chain reaction against internal transcribed spacer ribosomal DNA (rDNA) loci of the parasite followed by restriction fragment length polymorphism. Further, sequence analysis of 297 bp of ITS1-rDNA loci revealed the presence of L. major and Leishmania turanica in P. papatasi, and L. major in R. opimus. This is the first molecular report of L. major infection in both vectors (P. papatasi and P. caucasicus) and reservoir host (R. opimus) in this region. The results indicated that P. papatas was the primary vector of the disease and circulating the parasite between human and reservoirs, and P. caucasicus could be considered as a secondary vector. Further, our study showed that R. opimus is the most important host reservoir for maintenance of the parasite source in the area.

Background: Visceral leishmaniasis is caused by Leishmania infantum, transmitted to humans by bites of phlebotomine sand flies and is one of the most important public health problems in Iran. To identify the vector(s), an investigation was carried out in Bilesavar District, one of the important foci of the disease in Ardebil Province in northwestern Iran, during July-September 2008., Methods: Using sticky papers, 2,110 sand flies were collected from indoors (bedroom, guestroom, toilet and stable) and outdoors (wall cracks, crevices and animal burrows) and identified morphologically. Species-specific amplification of promastigotes revealed specific PCR products of L. infantum DNA., Results: SIX SAND FLY SPECIES WERE FOUND IN THE DISTRICT, INCLUDING: Phlebotomus perfiliewi transcaucasicus, P. papatasi, P. tobbi, P. sergenti, Sergentomyia dentata and S. sintoni. Phlebotomus perfiliewi transcaucasicus was the dominant species of the genus Phlebotomus (62.8%). Of 270 female dissected P. perfiliewi transcuacasicus, 4 (1.5%) were found naturally infected with promastigotes., Conclusion: Based on natural infections of P. perfiliewi transcaucasicus with L. infantum and the fact that it was the only species found infected with L. infantum, it seems, this sand fly could be the principal vector of visceral leishmaniasis in the region.

Shiraz district in south of Iran is a classical focus of Cutaneous leishmaniasis (CL) and previous research has consistently documented the etiologic agent to be Leishmania tropica and Leishmania major in urban and rural areas, respectively. However, none of the Phlebotomus sergenti, a known vector for L. tropica, of the region has been found infected. We report the first isolation of L. tropica from sandflies in urban community of southern part of Shiraz city. Parasite polymerase chain reaction (PCR) followed by restriction fragment length polymorphism (RFLP) and gene sequencing analyses indicate CL cases in this community were caused by either L. major or L. tropica. Sandflies of P. sergenti were infrequent, however, three out of 10 (30.0%) females captured in urban area were found infected with L. tropica. But, no human cases were found to be infected with L. tropica. Phlebotomus papatasi were found the most dominant and infected species where 41 out of 207 (20%) tested individuals harboring L. major in suburb area of the city. Patients have been lived in the suburb area of the city where people keep normally domestic animals in their houses which provide appropriate environment for completion of sandfly life cycle and expansion of CL disease in the region.

Background: A molecular survey was conducted to investigate the presence of pathogenic Borrelia persica species causing the tick borne relapsing fever (TBRF) in Takistan district Qazvin Province, western Iran., Methods: A number of 1021 soft ticks were collected from 31 villages including previously reported infected and none-infected TBRF cases and individually examined for the presence of B. persica DNA by conventional PCR targeting the 16S rRNA., Results: A total of 1021 soft ticks of three species of Ornithodouros tholozani (120: 11.75%), O. lahorensis (461: 45.15%) and Argas persicus (440: 43.1%) were collected and tested against Borrelia infection. Soft ticks were more prevalent (67%) in infected areas than none infected areas. The rate O. tholozani in infected areas was much greater (29 times) than none infected areas. Ninety seven percent of soft ticks in none infected areas were of O. tholozani. Sixteen (16.7%) ticks of tested (n=95) O. tholozani were infected with B. persica. Three (1.3%) out of 205 soft ticks of O. lahorensis were positive for Borrelia sp., and no infection was observed in A. persicus. TaqI RFLP analysis and sequence analysis of the positive PCR products showed the presence of B. persica. The RFLP analysis showed that the positive ticks of O. lahorensis were infected with unknown Borrelia species., Conclusion: This study showed that although there were no TBRF cases in Takisan, but still infected O. tholozani, the known vector of TBRF, presented in the region. Control measures needs to be fulfilled in Thakisan.

Leishmania infantum is the causative agent of infantile visceral leishmaniasis (IVL) in the Mediterranean Basin and, based on isoenzyme typing of the parasite isolated from dogs; this parasite was considered to predominate in the all foci of IVL in Iran. However, based on PCR detection and sequencing of parasite Cysteine Protease B (CPB), only one out of seven sandfly infections in Phlebotomus perfiliewi transcaucasicus was found to be L. infantum in the current investigation. The six other infections were haplotypes of Leishmania donovani, the causative agent of anthroponotic visceral leishmaniasis (AVL) in West Africa and India. The deduced amino acid of the L. donovani haplotype was found to be novel and the shortest CPB protein reported within the Leishmania spp. Circulation of both L. donovani and L. infantum by P. perfiliewi transcaucasicus, in addition to previous data indicating its ability to circulate L. tropica, suggests that this species, like other vectors of VL, is a permissive vector. Finding L. donovani infecting P. perfiliewi transcaucasicus in the area demands extensive and intensive typing of natural Leishmania infections in epidemiological investigations in Iran and the Mediterranean Basin in general.

The prevalence, host preference, and rate of Leishmania spp. infection of sand fly species are important parameters for incrimination of parasite vectors. We applied polymerase chain reaction (PCR)-based and enzyme-linked immunosorbent assay (ELISA) methods to detect Leishmania spp. parasites and blood meals within individual sand flies in the most important visceral leishmaniasis (VL) focus in northwestern Iran. Leishmania spp. minicircles (kinetoplast DNA) were found in 14 (0.9%) of 1,569 female specimens. Sequence analysis of 650 basepairs of an internal transcribed spacer ribosomal DNA gene identified L. infantum/L. donovani in 12 specimens and L. adleri-like parasites in 2 specimens. Nine (64.3%) of 14 of the Leishmania spp.-positive sand flies were Phlebotomus perfeliewi transcaucasicus. Blood meal identification of host DNA within sand flies by PCR-based and ELISA methods showed that 30% and 28%, respectively, were positive for human blood. Results of this study showed that P. perfeliewi transcaucasicus is the most prevalent, infected, and anthropophagic sand fly and plays a major role in VL transmission in the region studied.