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Background: Visceral leishmaniasis (VL) or kala-azar is a parasitic disease caused by the species of Leishmania donovani complex. Mediterranean type of the disease is endemic in some parts of Iran and more than 95% of cases were reported in children up to 12 years of age. This study was performed to determine the seroprevalence of VL in the rural areas of the Dashti district from Bushehr province. Materials and Methods: In this cross-sectional study, a randomized cluster sampling method was used for the collection of blood samples from children up to 12 years old from rural areas of Dashti district. Before sampling; a questionnaire was filled out for each case. All the collected blood samples were examined after the serum separating by Direct Agglutination Test (DAT) for detection of anti-Leishmania infantum antibodies. The cutoff titers of ≥1: 3200 with specific clinical features were supposed to be considered as VL. Results: Altogether, 24 out of 1221 (1.96%) blood samples showed titers between 1:800 and 1:1600 which considered as suspicious cases. None of the suspicious cases had a history of kala-azar. None of 1221 collected blood samples showed anti Leishmania infantum (L. infantum) at titer ≥1:3200. Conclusion: This study confirms the circulation of L. infantum in Dashti district and highlights the sporadic pattern of VL in the studied areas which necessitates the surveillance system to be monitored by health authorities. [ABSTRACT FROM AUTHOR]

Background: Visceral leishmaniasis (VL), caused by Leishmania infantum (L. infantum), is a life-threatening vector-borne parasitic disease is distributed in some parts of the world. The disease is endemic in some parts of Iran. This study was aimed to determine the seroprevalence of VL among children and domestic dogs (as a reservoir of the parasite) in Dehloran, west of Iran. Materials and Methods: This cross-sectional study was carried out in Dehloran County. The blood samples of 872 children up to 12 years old and 52 dogs were collected from 10 villages of Dehloran using randomlyclustered sampling method. Sera were separated from all peripheral blood samples and tested by direct agglutination test (DAT). Anti-Leishmania infantum antibodies at titers of ≥1:800 and ≥1:80 were considered as Leishmania infantum infection in human and dog, respectively. Results: In general, among 872 human samples, 1.03% of samples had anti-Leishmania antibody with 1:1600 titers and 1.26% had 1:800 titers. In addition, from 52 dog samples, 21.15% of dogs had a titer of ≥1:320 and 25% had 1:80 and 1:160 titers. Conclusion: Our findings indicate that the seropositive dogs in the studied areas are considerable and L. infantum may be circulated between human and domestic dog in the studied area. Further study of isolation and molecular identification of Leishmania spp. is recommended. [ABSTRACT FROM AUTHOR]

Objectives: This study was aimed to evaluate the role of toxoplasmosis in etiology of abortion and stillbirth based on molecular and serological techniques.Material and Methods: A total of 110 pregnant women with abortion and stillbirth were enrolled as the case group, and 110 pregnant women with normal delivery were enrolled as the control group. Serological and molecular detections of Toxoplasma gondii were assessed by ELISA and PCR methods.Results: The seroprevalence of IgG was 25.5% in the case group (26.8% in abortion and 21.4% in stillbirth) and 26.4% in the control group. IgM seropositivity was detected in 2.7% of the case group (3.6% in abortion and 0% in stillbirth) and 0.9% of the control group (p = 0.37). Toxoplasma gondii DNA was detected in 6.4% of the case group (7.3% in abortion and 3.6% in stillbirth) and 1.8% of the control group by PCR (p = 0.17). The major risk factor of congenital toxoplasmosis was the history of eating undercooked meat (p = 0.06).Conclusion: Results of this study revealed that the rate of PCR positive in women with abortion and stillbirth was 3.7 times higher than that in normal delivery, but the difference was not statistically significant. These findings suggest that toxoplasmosis can be involved in etiology of abortion and stillbirth. [ABSTRACT FROM AUTHOR]

Background: Free-living amoebae such as Acanthamoeba species may act as carriers of Cryptosporidium and Toxoplasma oocysts, thus, may play an important role in the water-borne transmission of these parasites. In the present study, a loop mediated isothermal amplification (LAMP) method for detection of Toxoplasma and a PCR assay were developed for investigation of Acanthamoeba in environmental water samples. Methods: A total of 34 samples were collected from the surface water in Guilan Province. Water samples were filtrated with membrane filters and followed by DNA extraction. PCR and LAMP methods used for detection of the protozoan parasites Acanthamoeba and Toxoplasma respectively. Results: Totally 30 and 2 of 34 samples were positive for Acanthamoeba and Toxoplasma oocysts respectively. Two samples were positive for both investigated parasites. Conclusion: The investigated water supplies, are contaminated by Toxoplasma and Acanthamoeba (oo)cystes. Acanthamoeba may play an important role in water-borne transmission of Toxoplasma in the study area. For the first time in Iran, protocol of LAMP method was used effectively for the detection of Toxoplasma in surface water samples in Iran. [ABSTRACT FROM AUTHOR]

Background: Haptoglobin (Hp) is a plasma α2-sialoglycoprotein that contains alpha and beta chains. It displays in three common phenotypes, Hp1-1, Hp2-1, and Hp2-2. Proteins expressed by polymorphic genes have grossly different molecular sizes resulting in different diffusion rates in the brain. Haptoglobin expressed by the Hp2-2 genotype has lower hemoglobin-binding capacity than Hp1-1 or Hp2-1 and is associated with idiopathic generalized epilepsy. Methods: To determine polymorphism in haptoglobin genes in patients with idiopathic generalized tonic-clonic seizures, 42 men, 42 women, and 50 controls were selected for this study. Genomic DNA was extracted from blood and studied by polymerase chain reactions (PCR). Results: The amplified fragments for the Hp1-1 and Hp2-2 genotypes were 1757 and 3481 base pairs (bp) respectively, and the Hp2-1 genotype had both fragments, in addition to a 349-bp fragment. The distribution of the three major Hp phenotypes in epilepsy patients was 28.6 (1-1), 38.1 (2-1), and 33.3% (2-2) in the men, and 31 (1-1), 40.5 (2-1), and 28.6% (2-2) in the women. The distribution of Hp genotypes in controls was 22 (1-1), 40 (2-1), and 38% (2-2). Conclusion: We show that all Hp genotypes participate in idiopathic generalized epilepsy. [ABSTRACT FROM AUTHOR]

Background: The present study was carried out to investigate the accurate status of ovine Theileria infection in sheep from Ahvaz and surrounding region, a tropical area southwest Iran. Methods:A PCR-RFLP method based on 18S ribosomal RNA gene was designed which could detect and differentiate Theileria and Babesia spp. and also differentiate main Theileria species in sheep at the same time. 119 sheep blood samples were collected from Ahvaz and surroundings. Results: Microscopic examination of blood smears revealed 69.7% (83/119) infection with Theileria spp. Of the total samples subjected to PCR, 89% (106/119) were found to be positive, all of which were identified as Theileria by RFLP analysis using enzyme Hind II. In enzymatic digestion of PCR products by Vsp I, 91.5% (97/106) of Theileria positive samples were identified as T. ovis while mixed Theileria infections were found in 9 samples. The samples with mixed infections were analyzed with an additional nested PCR-RFLP method, by HpaII enzyme digestion. 3 samples with T. lestoquardi infection, 1 sample with T. ovis and T. annulata, 1 sample with T. lestoquardi and T. annulata, and 4 samples with T. ovis, T. lestoquardi and T. annulata mixed infections were detected. Conclusion: Ovine theileriosis caused by T. ovis is highly prevalent in southwest Iran while T. lestoquardi and T. annulata infection can be detected in a lesser proportion of sheep in this region. The new PCR-RFLP method that was designed in this study, can serve as a beneficial diagnostic tool, especially in T. ovis prevalent regions. [ABSTRACT FROM AUTHOR]

Abstract: Objective: To identify Leishmania using PCR. Methods: This study was conducted from April 2009 to March 2011 in order to identify Leishmania species in a new endemic area of CL in Lorestan, Iran. Samples were taken from 62 patients that referred to the health centers in different cities of Lorestan province, the presence of Leishmania was confirmed using direct smear and then grown in NNN media and mass cultured in RPMI 1 640 medium supplemented with 10% heat-inactivated fetal bovine serum. DNA was extracted from cultured promastigotes and used in ITS-PCR. Results: 45(72.6%) samples out of 62 showed a band in the range of 485 bp and 17 (27.4%) with a band in the range of 626 bp which were similar to standard strains of Leishmania tropica(L. tropica) and Leishmania major(L. major), respectively. 50 (65.80%) of samples were collected from people with no history of travel in at least a year prior to the onset which shows that indigenous source of infection. Conclusions: Since the vector and reservoir of the two species are different, so precise and extensive control and prevention methods should be designed and carried out. [Copyright &y& Elsevier]

Background Cryptosporidium and Giardia in water supplies is acknowledged as a public health problem. In the present study, we applied immunofluorescence assay (IFA), PCR and loop-mediated isothermal amplification (LAMP) for the detection of the two protozoa. Methods Over a period of 12 months, surface water samples were collected from two rivers in the north of Iran, and filtrated by 142 mm membrane filters. At each sampling point 10 L water were used for IFT and the10 L were analysed using molecular methods. Results In 15/40 samples, (oo)cysts were detected by one of the IFA, PCR or LAMP methods. Five samples that were Cryptosporidium-negative by IFA were positive by LAMP. A total of 10 out of 13 samples that were Giardia-positive by IFA were also positive by PCR. IFA revealed high levels of Giardia, with 1–1800 cysts and 1–16 Cryptosporidium oocysts detected per 10 L. Conclusion The study reveals that the investigated water supplies were contaminated by Cryptosporidium and Giardia. The LAMP assay has advantages for detection and screening of these protozoa at relatively low concentration in water samples. The three assays applied are complimentary but no single one will give the true prevalence of these parasites in surface water samples. However, each method has its own advantages and disadvantages dependent of the aim and the study design; a combination of detection methods should be applied to discover whether water is, or is not, contaminated with (oo)cysts. [ABSTRACT FROM AUTHOR]

Background: Leishmaniasis is a group of diseases that are created by intracellular parasites of Leishmania. Cutaneous leishmaniasis is considered as one of the health problems in some provinces of Iran. Methods: In this study, a total of 178 Giemsa-stained slides from confirmed cases of cutaneous leishmaniasis were examined. The slides were prepared from the patients with cutaneous leishmaniasis that referred to health centers and infected during the epidemic of cutaneous leishmaniasis in Poldokhtar city, Lorestan Province, Iran in 2006.Genomic DNA from each slide was extracted. After DNA extraction, ITS-PCR was used. Results: Out of 178 slides, 129 (72.47%) samples had a band in the range of 485 bp and 49 (27.53%) samples 626 bp that matched L. tropica and L. major standard samples, respectively. Conclusion: This study showed that Leishmania DNA could be efficiently extracted and amplified even from old Giemsa-stained microscopic slides that were stored more than 6 yr. In this study was shown that both L. tropica and L. major species exist in Lorestan Province. [ABSTRACT FROM AUTHOR]

Infection with human papiloma virus (HPV) is the most frequent sexually transmitted disease worldwide. HPV types 16, 18, 31 and 33 are considered as the most important types in the cervical cancer. This study was undertaken on 64 samples of archival cervical carcinoma pathologic to assess the rate of HPV infection (HPV16,18) in cervical carcinoma among Iranian patients. HPV DNA was detected by polymerase chain reaction (PCR) and typing by restriction fragment length polymorphism (RFLP) analysis. The total prevalence of HPV in this study (HPV16,18) for all cases was 59.4% (38/64). HPV type 16 was the most common one (22/64, 34%) followed by HPV type 18 (16/64, 25%). On the basis of the rate of HPV (16,18) which were detected in squamous cell carcinoma and adenocarcinoma, only women with HPV18 infection showed a statistically significant risk for development of cervical cancer (P=0.019) while P value for HPV16 was 0.47. [ABSTRACT FROM AUTHOR]

Summary: We investigated the prevalence of intestinal protozoan parasites in patients with gastrointestinal complaints in medical centers in Zahedan, Iran. A total of 1562 stool samples was examined from July 2004 to January 2006 using microscopy (direct smear, formalin-ether concentration), xenic culture and PCR techniques. Four hundred and twenty-seven (27.3%) of the patients were infected with one or more intestinal parasites. Giardia lamblia (10.1%), Entamoeba coli (10%), E. hartmanni (1.7%), Blastocystis hominis (2.2%), Chilomastix mesnili (1.7%), Trichomonas hominis (0.7%), E. histolytica/E. dispar (0.51%) and Iodamoeba butschlii (0.45%) were the most prevalent protozoa detected with microscopy. Of the eight microscopy-positive E. histolytica/E. dispar samples, six were identified as E. dispar by PCR/gel electrophoresis, whereas E. histolytica was not detected at all. Although Zahedan is an area with poor hygiene located in a tropical area near the border of Pakistan and Afghanistan, the prevalence of E. histolytica and E. dispar here compared with other parasites and infectious diseases is unexpectedly low. [Copyright &y& Elsevier]

Trichomonas vaginalis is a human urogenital pathogen that causes trichomoniasis, the most common nonviral, parasitic sexually transmitted infection in the world. At present, little is known regarding the degree of strain variability of T. vaginalis . A classification method for T. vaginalis strains would be a useful tool in the study of the epidemiology, drug resistance, pathogenesis and transmission of T. vaginalis . Eight different types of actin genes have been identified by PCR-RFLP in T. vaginalis ; the purpose of this study is to determine the genotypes of this parasite in Karaj city, Iran. Forty-five clinical T. vaginalis isolates from vaginal secretions and urine sediment were collected from Karaj city from 2012 through 2014. DNA was extracted and the actin gene was amplified by nested-PCR; all samples were positive. To determine the genetic differences, sequencing on seven samples was conducted. Then, all PCR products were digested with HindII, MseI, and RsaI restriction enzymes. Of 45 isolates, 23 samples (51.1%) were of actin genotype G, 11 samples (24.4%) of genotype E, six samples (13.3%) of genotype H, three samples (6.6%) of genotype I, and two samples (4.4%) were mixed genotypes of G and E. Genetic diversity of T. vaginalis isolates is notable. The actin genotype G may be the dominant genotype in Karaj city, Iran. [ABSTRACT FROM AUTHOR]

Abstract: This study has identified and characterized the structure of locus D-A, a noncoding short tandem repeat (STR) region, also known as locus 1–2, in Iranian Entamoeba dispar isolates. This polymorphic locus has been shown to be potentially useful in investigating the molecular epidemiology of Entamoeba histolytica and E. dispar. The genetic polymorphisms in locus D-A in 28 isolates of E. dispar from three different geographic regions of Iran were distinguished using PCR and sequencing, and the results were compared with the E. dispar gene sequences available in GenBank. In all microscopy-positive E. histolytica/E. dispar samples, PCR with species-specific primers was used to amplify a 477–531bp product, identifying the samples that had E. dispar. Analysis of the sequences revealed a remarkable degree of genetic diversity with regard to size, number and organization of the repeat units among the E. dispar isolates. The sequenced products showed 12 novel E. dispar genotypes, which have been submitted to the GenBank/EMBL/DDBJ database under accession numbers AB354125–AB354136 AB354125 AB354126 AB354127 AB354128 AB354129 AB354130 AB354131 AB354132 AB354133 AB354134 AB354135 AB354136 . [Copyright &y& Elsevier]

After half a century, Trichinella infection has been reported in humans following ingestion of wild boar meat hunted in Northern Iran. We have performed a cross-sectional study to evaluate the prevalence of Trichinella spp. infections in wild boar and prevalence of anti- Trichinella antibodies among at-risk individuals for the first time in Iran. Muscle and sera samples were collected from 79 wild boars and 364 at-risk individuals. Trichinella infection has been investigated by artificial digestion and molecular identification (in wild boar muscles) and by serology (in humans). Trichinella larvae were isolated from three wild boars (3.7%; 95% CI, 2.9–4.3). The isolated larvae were identified as T. britovi using CO1 and 5S rRNA gene primers amplification. The percent identity regarding to 5S rRNA gene (98.7–100%) and divergence (0–1.9%) further confirmed the isolates as T. britovi . Of the 364 participants, anti- Trichinella IgG were detected in 8 (2.2%; CI 95%, 1.9–2.4). Risk factors associated with Trichinella infection seropositivity in humans, were hunter being (OR, 13.5; 95% CI, 3.1–59.4; P = 0.003) and high consumption (more than 7 time in a year) of wild boar meat (OR, 17.5; 95% CI, 3.2–93.6; P < 0.001). In conclusion, results of this study emphasized that consumption of wild boar meat could be important source of human trichinellosis as a completely neglected infection disease in Iran. We suggest that the additional studies should be performed in different parts of Iran to further clarify the prevalence of trichinellosis in wild animals to guide the development of appropriate public health interventions. [ABSTRACT FROM AUTHOR]

Abstract: The diagnosis of visceral leishmaniasis (VL) in humans and animal reservoir hosts is difficult, particularly in rural areas where the disease is endemic and laboratory facilities are limited. This study aimed to develop a latex agglutination test (LAT) for the rapid detection of anti-Leishmania antibodies against the A2 antigen derived from the amastigote form as well as those against crude antigens derived from the promastigote form of an Iranian strain of Leishmania (Leishmania) infantum. The A2 antigen (42–100kDa) was prepared from the amastigote form of L. infantum, purified with electroelution and compared with the crude antigen from the promastigote form of L. infantum. Both antigens showed appropriate intensity reactions, were selected using dot blotting of positive and negative pooled sera and used to sensitize 0.9-μm latex beads. The tests were carried out on sera from 43 symptomatic, human patients with VL confirmed by parasitological examination and direct agglutination test (DAT), 30 healthy controls and 32 patients with other infections but without VL. Canine sera were collected from 63 domestic dogs with VL confirmed using parasitological examinations and DAT and 31 healthy dogs from areas non-endemic for VL. Compared with the controls, human sera from DAT-confirmed patients yielded a sensitivity of 88.4% (95% CI, 82.1–94.5%) and specificity of 93.5% (95% CI, 87.0–99.7%) on A2-LAT (amastigote) when 1:3200 was used as the cut-off titre. A good degree of agreement was found between A2-LAT and DAT (0.914). LAT required 3–5min to complete, versus the 12–18h needed for DAT. Compared with the controls, A2-LAT of canine sera from DAT-confirmed cases yielded a sensitivity of 95.2% (95% CI, 95.0–95.4%) and specificity of 100% (95% CI 100%) when 1:320 was used as the cut-off titre. A good degree of agreement was found between A2-LAT and DAT (0.968). Similarly, the sensitivity and specificity of Pro.-LAT (promastigote) was calculated to be 88.4% and 91.9%, respectively for human sera and 96.8% and 90.3%, respectively for canine sera. No statistically significant differences were observed between A2-LAT and Pro.-LAT for the detection of human and canine L. infantum infections. In conclusion, A2-LAT and Pro.-LAT could be used in parallel to screen for L. infantum infections in humans and dogs in areas endemic for VL in Iran. [Copyright &y& Elsevier]

Abstract: Clinically infected dogs have been identified as the main reservoir hosts of visceral leishmaniasis (VL) caused by Leishmania infantum in the Mediterranean region. The objective of this study was to determine the potential of asymptomatic infected dogs compared with symptomatic ones as a source of L. infantum infection to golden hamster. For this purpose, anti-Leishmania antibodies were detected with direct agglutination test (DAT) in 13 symptomatic (7 seropositive=≥1:320) and 53 asymptomatic (9 seropositive=≥1:320 and 44 seronegative= <1:320) ownership dogs. DNA of Leishmania sp. was extracted from skin and peripheral blood tissues of each dog and tested by PCR. Sixty-six Syrian golden hamsters (Mesocricetus auratus) were used for the determination of infectivity and pathogenicity of L. infantum, isolated from the dogs. We used the internal transcribed spacer 2 (ITS 2) rDNA sequence analysis. The results showed that 22 and 11 out of 66 inoculated golden hamsters were positive by PCR and parasitological examinations, respectively. From 22 PCR positive hamsters, 17 were related to asymptomatic dogs and 5 were from symptomatic ones. There was no significant difference between symptomatic and asymptomatic dogs in producing Leishmania infection in the susceptible animal model (P =0.66). Smears and cultures of 5 dogs from 13 symptomatic dogs (38.5%) and 6 dogs from 53 asymptomatic ones (11.3%) were found to be positive at parasitological examination. All the L. infantum isolates from symptomatic and asymptomatic dogs were similar in sequencing. In conclusion, asymptomatic infected dogs as well as symptomatic ones can harbor L. infantum in their blood and skins which are virulent and infectious for inoculated golden hamster. [Copyright &y& Elsevier]