Intestinal parasitic infections frequency in referred patients to a large teaching hospital, Khuzestan, Southwest, Iran, 2017.

Intestinal parasitic infections (IPIs) are among the most important infectious diseases in Iran. A cross sectional study was designed to determine frequency of intestinal parasites among referrals to a large teaching hospital in Khuzestan, Southwest of Iran, 2017. A total number of 5613 stool samples were examined through direct smear and formalin-ether concentration methods to detect possible parasitic infections. Samples consisted of 2643 (47.09%) male and 2970 (52.91%) female. A total of 1468 (26.15%) samples were positive (13.11% male and 13.4% female) and 4145 (73.85%) were negative. The results also showed that 255 of samples had more than one type of parasite (mix infections). Counting single and mix parasite infections, the total number of positive cases reached to 1723. Helminthes parasites were present in 12 (0.7%) cases, while intestinal protozoan parasites were in 1711 (99.3%) cases. Almost equally, pathogenic and nonpathogenic parasites infected 860 (49.91%) and 863 (50.09%) of patients, respectively. The frequency for helminthes was determined at 0.52% with Hymenolepis nana and Enterobius vermicularis however, Giardia lamblia in 38.54% and Entamoeba histolytica/dispar at 10.68% were concluded as protozoa elements. The IPIs frequency was recorded in female and male patients at 49.16% and 50.14%, respectively. According to the current results the infection rate of intestinal parasites has been significantly reduced especially for helminths infections in this region possibly due to public attention to health issues such as; increased awareness of people, improvement of sanitation, seasonal variations, health education and personal hygiene.

Intestinal parasitic infections frequency in referred patients to a large teaching hospital, Khuzestan, Southwest, Iran, 2017

@#Intestinal parasitic infections (IPIs) are among the most important infectious diseases in Iran. A cross sectional study was designed to determine frequency of intestinal parasites among referrals to a large teaching hospital in Khuzestan, Southwest of Iran, 2017. A total number of 5613 stool samples were examined through direct smear and formalin-ether concentration methods to detect possible parasitic infections. Samples consisted of 2643 (47.09%) male and 2970 (52.91%) female. A total of 1468 (26.15%) samples were positive (13.11% male and 13.4% female) and 4145 (73.85%) were negative. The results also showed that 255 of samples had more than one type of parasite (mix infections). Counting single and mix parasite infections, the total number of positive cases reached to 1723. Helminthes parasites were present in 12 (0.7%) cases, while intestinal protozoan parasites were in 1711 (99.3%) cases. Almost equally, pathogenic and nonpathogenic parasites infected 860 (49.91%) and 863 (50.09%) of patients, respectively. The frequency for helminthes was determined at 0.52% with Hymenolepis nana and Enterobius vermicularis however, Giardia lamblia in 38.54% and Entamoeba histolytica/dispar at 10.68% were concluded as protozoa elements. The IPIs frequency was recorded in female and male patients at 49.16% and 50.14%, respectively. According to the current results the infection rate of intestinal parasites has been significantly reduced especially for helminths infections in this region possibly due to public attention to health issues such as; increased awareness of people, improvement of sanitation, seasonal variations, health education and personal hygiene.

The occurrence of Toxocara species in naturally infected broiler chickens revealed by molecular approaches.

Consuming raw and undercooked meat is known to enhance the risk of human toxocariasis because Toxocara species have a wide range of paratenic hosts, including chickens. The aim of this study was to identify species of Toxocara in naturally infected broiler chickens using molecular approaches. A polymerase chain reaction (PCR) method was used for the differentiation of Toxocara canis and Toxocara cati larvae recovered from tissues and organs, and identified by microscopic observations. Thirty-three 35- to 47-day-old broiler chickens were used for examination of Toxocara larvae. The duodenum, liver, lungs, heart, kidneys, skeletal muscles and brain of each chicken were examined using the pepsin method, and DNA from each tissue was extracted as the template for PCR assay. The findings revealed that 5 of 33 (15.2%) broiler chickens were infected with Toxocara larvae. Larvae were recovered from the liver (n = 19), duodenum (n = 8), skeletal muscles (n = 8) and brain (n = 2) of broiler chickens naturally infected with Toxocara spp. The results showed that the frequencies of the species in the chickens were T. canis larvae (n = 5, 83.3%) and T. cati larvae (n = 1, 16.7%). Our data from the present study demonstrated the importance of broiler chickens as a paratenic host for the parasite's life cycle in the environment. The implementation of DNA amplification as a routine diagnostic technique is a specific and alternative method for identification of Toxocara larvae, and allowed the observation of specific species under field conditions within the locations where broiler chickens are typically raised and exposed to Toxocara spp. eggs or larvae.

Correlation between clinical responses with the drugsusceptibility of parasites in Iranian cutaneous leishmaniasis caused by Leishmania major.

Reviews have shown increasing number of Iranian patients with cutaneous leishmaniasis (CL) who are unresponsive to pentavalent antimonial compounds such as meglumine antimoniate (Glucantime, MA). The present investigation aims to determine the correlation between clinical responses (healing, or non-healing) with susceptibility of Leishmania parasites to glucantime. Initially, in vitro susceptibility of Leishmania parasites was carried out on 93 isolates using macrophage models. Identification of these species was also performed by molecular methods including Nested-PCR and PCR-RFLP. The f indicated that total isolated were L. major. A significant association between the clinical outcome and the in vitro effective concentration 50% (EC50) values was observed. Leishmania derived from patients with non-healing lesions had EC50 values at least 3-fold higher than parasites isolated from lesions of healing patients. By molecular methods, patterns for both sensitive and resistant samples demonstrated restriction band which is related to L. major. The obtained findings in the present study demonstrated that MA-resistant L. major field isolates are now frequent in Iran. Such studies help to find strategies for rapidly diagnosing resistance in order to improve the clinical management of CL.

Evidence that the Echinococcus granulosus G6 genotype has an affinity for the brain in humans.

The present study investigates the molecular characteristics of cerebral Echinococcus cysts. A total of 10 specimens of cerebral Echinococcus cysts, including six formalin-fixed paraffin blocks and four intact cerebral cysts, were used for this study. The target DNA was successfully amplified from eight samples and sequenced. BLAST analysis indicated that sequenced isolates belong to the Echinococcus granulosus (G6) genotype. All of the eight sampled brain cysts belonged to the G6 genotype, while all of the eight liver cysts belonged to G1. This is a strong indication that G6 has a higher affinity for the human brain than G1.

Lysozyme activity in the plasma of rodents infected with their homologous trypanosomes.

BACKGROUND: In this study the concentration of lysozyme in blood plasma of Microtus agrestis, Clethrinomys glareolus, Apodemus sylvaticus, BK rats and outbred white mice before and after infection with culture forms of Trypanosoma microti, T, evotomys, T. grosi, T. lewisi and T. musculi respectively was measured. METHODS: Blood samples of rodents, Microtus agrestis, Clethrionomys glareolus, Apodemus sylvaticus, BK rats and outbred mice infected with T. microti, T. evotomys, T. grosi, T. lewisi and T. musculi respectively were collected in heparinized micro- tubes immediately before inoculation and 3, 6, 12, 24, 48, 96 and more than 400 days after intra- perituneal inoculation with 5×10(5)of their homologous trypanosome parasites of which more than half were metacyclic trypomastigote in 0.2 ml of culture medium. Micro- tubes were centrifuged and plasma samples were separated and the lysozyme activity was measured by the agar method. RESULTS: Levels of lysozyme rose rapidly three to six days after the inoculation to ten to twenty than their pre- infection levels. They then gradually decreased, although after more than one year they were still two to ten folds higher than controls. The highest level measured occurred in rats infected with T. lewisi and the lowest in A. sylvaticus infected with T. grosi. After one year the highest concentration of lysozyme was in mice infected with T. musculi and lowest in A. sylvaticus. CONCLUSION: Persistent enhanced lysozyme levels may prevent re- infection with trypanosomes.

Seroprevalence of toxocariasis in hypereosinophilic individuals in Ahwaz, south-western Iran.

Eosinophilia in human peripheral blood is caused by different agents, including toxocariasis. The present study aimed to evaluate the prevalence of toxocariasis in hypereosinophilic individuals in the city of Ahwaz, located in south-western Iran, using enzyme-linked immunosorbent assay and Western blot techniques. Serum samples were examined from 100 individuals with peripheral blood eosinophilia and also from another 100 individuals without eosinophilia as the control group. In hypereosinophilic individuals seroprevalence antibodies against Toxocara were found in 19 (19%), of whom 12 (63.15%) were female and 7 (36.85%) were male. Positive sera were subsequently confirmed by Western blot. All of the observed bands ranged from 24 to 100 kDa. Antibodies against Toxocara were found in 1% of the control group, but were not confirmed by Western blot. The results showed significant differences between the frequency of infection within age and gender (P < 0.05); the highest prevalence of infection was observed in adults. Differences between the hypereosinophilic and healthy individuals, in terms of Toxocara infection frequency, also proved significant (P < 0.05).The present study thus confirmed the significant prevalence of toxocariasis as a hygienic problem among hypereosinophilic individuals in this area. It is, therefore, necessary to examine these individuals for toxocariasis.

The PCR-based detection and identification of the parasites causing human cutaneous leishmaniasis in the Iranian city of Ahvaz.

In Iran, Leishmania major or L. tropica cause almost all of the human cutaneous leishmaniasis (CL). Unfortunately, the detection methods frequently used for CL (the microscopical examination of direct smears or the culture of biopsies) are not very sensitive and the Leishmania species causing each case of CL in Iran is usually only tentatively identified from extrinsic factors, such as the case's clinical manifestations and region of residence. Recently, however, a nested PCR that targets the parasites' kinetoplast DNA has been used in the city of Ahvaz (the capital of the province of Khouzestan, in south-western Iran) to confirm the microscopical diagnosis of CL and to identify the causative parasites, to species level. Smears from the lesions on 100 suspected cases of CL were fixed, stained with Wright's eosin-methylene blue, and checked for amastigotes under a light microscope. Scrapings from the same smears were then tested for leishmanial DNA, using a nested PCR that allows the DNA from L. tropica to be identified and distinguished from that of L. major. The 100 smears investigated were all found amastigote-positive by microscopy and PCR-positive for either L. major DNA (97 smears) or L. tropica DNA (three smears). The predominant species causing CL in Ahvaz is therefore L. major.

Diagnosis and identification of Leishmania spp. from Giemsa-stained slides, by real-time PCR and melting curve analysis in south-west of Iran.

BACKGROUND: The aim of present study was describing a real-time PCR assay for the diagnosis and direct identification of Leishmania species on Giemsa-stained slides in south-west of Iran. MATERIALS AND METHODS: Altogether, 102 Giemsa-stained slides were collected from different part of south-west of Iran between 2008 and 2011. All the Giemsa-stained slides were examined under light microscope. After DNA extraction, real-time PCR amplification and detection were conducted with fluorescent SYBR Green I. For identification, PCR products were analysed with melting curve analysis. RESULTS: One hundred and two archived slides from suspected lesion examined by microscopy and real-time PCR. The sensitivity of the real-time PCR on Giemsa-stained slid was 98% (96/102). The melting curve analysis (T(m)) were 88·3±0·2°C for L. tropica (MHOM/IR/02/Mash10), 86·5±0·2°C for L. major (MHOM/IR/75/ER) and 89·4±0·3°C for L. infantum (MCAN/IR/97/LON 49), respectively. CONCLUSION: This study is first report in use of real-time PCR for diagnosis and identification of Leishmania spp. in Iran. Up to now, in Iran, the majority of identification of Leishmania species is restriction fragment length polymorphism (PCR-RFLP) of ITS1 and kinetoplast DNA. Our data showed that Giemsa-stained slides that were stored more than 3 years, can be use for Leishmania DNA extraction and amplification by real-time PCR. Compared to conventional PCR-based methods, the real-time PCR is extremely rapid with results and more samples can be processed at one time.

Mini-exon genotyping of leishmania species in khuzestan province, southwest iran.

BACKGROUND: Leishmaniasis is a protozoan disease cause by Leishmania genus. Anthroponotic and zoonotic cutaneous leishmaniasis are endemic in Iran. The aim of this study was to identify the causative agent of cutaneous leishmaniasis by mini-exon gene in five regions of Khuzestan Province, southwest of Iran. METHODS: From 2007 to 2008 in this cross-sectional study, cutaneous samples were collected from patients referred to Health Centers and Hospitals of the Khuzestan Province for cutaneous leishmaniasis diagnosis and cultured in Novy-MacNeal-Nicolle (NNN) and RPMI 1640. The propagated promastigotes were harvested and Leishmania species of cutaneous leishmaniasis were identified by RFLP and DNA sequencing of the PCR generated fragments. RESULTS: L. major and L. tropica were the causative agents of cutaneous leishmaniasis by predominantly of L. major species. The alignment of the mini-exon sequencing isolates with reported sequencing of L. major and L. tropica revealed 92%-99% identity. CONCLUSION: Our study showed that mini-exon PCR-RFLP was useful method to identify the causative species of cutaneous leishmaniasis.

Human cystic echinococcosis in nomads of south-west Islamic Republic of Iran.

We investigated the prevalence of human CE among nomadic communities in 4 areas of Khuzestan province: Behbahan, Shoush, Masjed Soleiman and Izeh. Blood samples from 3446 individuals from 700 randomly selected families were examined for detection of antibody against Echinococcus granuIosus. Family members were interviewed to assess possible risk factors for infection such as age, sex, dog ownership. The prevalence of CE was 13.8%: 1.9% in Behbahan, 12.4% in Shoush, 17.3% in Masjed Soleiman and 18.2% in Izeh. These differences were statistically significant. There was no significant association between CE seropositivity and age, sex and dog ownership.

Human cystic echinococcosis in nomads of south-west Islamic Republic of Iran

Cystic echinococcosis [CE] is a widespread helminth zoonosis, especially in rural areas. We investigated the prevalence of human CE among nomadic communities in 4 areas of Khuzestan province: Behbahan, Shoush, Masjed Soleiman and Izeh. Blood samples from 3446 individuals from 700 randomly selected families were examined for detection of antibody against Echinococcus granulosus. Family members were interviewed to assess possible risk factors for infection such as age, sex, dog ownership. The prevalence of CE was 13.8%: 1.9% in Behbahan, 12.4% in Shoush, 17.3% in Masjed Soleiman and 18.2% in Izeh. These differences were statistically significant. There was no significant association between CE seropositivity and age, sex and dog ownership

Serotyping of group A rotaviruses in Egyptian neonates and infants less than 1 year old with acute diarrhea.

Group A human rotavirus G serotypes were detected in stool specimens from neonates and infants with and without acute diarrhea in Cairo by using monoclonal antibodies in an enzyme-linked immunosorbent assay. Serotypes G1 and G4 predominated in all age groups. Mixed (G1 plus G4) and nontypeable specimens represented 16.1 and 38.7% of the total number serotyped, respectively.

PIP: Group A human rotavirus (HRV) is the most common cause of acute gastroenteritis among infants worldwide. 7 G serotypes of group A HRV have thus far been identified by neutralization tests, of which 4, serotypes G1 through G4, have a global distribution. Enzyme immunoassay (EIA) was used to detect the relative frequency and temporal distribution of HRV G serotypes 1-4 among the community of neonates and infants with and without acute diarrhea who attended Cairo University Children's Hospital between August 1992 and October 1993. Fecal samples were collected from 20 neonates and 109 infants under age 1 year with acute diarrhea and from 20 neonates and 30 infants without acute diarrhea. Samples were then tested for the presence of rotavirus antigen using a commercial EIA. Rotavirus was identified in 64 of the 180 neonates and infants; in 15 neonates with diarrhea, 6 neonates without diarrhea, and 43 infants aged 1-12 months. Infection peaked between August and December. Serotypes G1 and G4 predominated in all age groups. Mixed (G1 and G4) and nontypeable specimens represented 16.1% and 38.7% of the total number serotyped, respectively.

In vitro cultivation of Trypanosoma acomys: production of insect stages and bloodstream forms.

When Trypanosoma acomys bloodstream forms were cultivated at 37 degrees C in Schneider's Drosophila medium supplemented with 20% (v/v) heat-inactivated foetal calf serum (FCS), with Microtus agrestis embryonic fibroblasts in RPMI 1640 medium supplemented with 20% FCS or in Baltz's medium supplemented with 10% FCS, the parasites transformed and largely remained as epimastigotes. Epimastigotes were also usually the commonest stage observed when the parasites were co-cultivated with a mosquito cell line at 27 degrees C. However, if these cultures were initiated with the supernatant suspensions from fibroblast cultures that had been cryopreserved, trypomastigotes, including bloodstream-like forms, were the predominant stage for the first 4 days of culture. It is suggested that the glycerol supplement or the temperature changes stimulated this unusual morphogenesis. At 27 degrees C, T. acomys was incapable of multiplying and died when cultured in fresh Schneider's Drosophila medium supplemented with 20% FCS, but co-cultivation with the mosquito cell lines or cultivation in cell-free supernatants from 1-week-old mosquito cell cultures was successful at this temperature; most of the parasites multiplied as epimastigotes.

Oral transmission of trypanosomes of the subgenus Herpetosoma from small mammals.

The rodents Microtus agrestis, Clethrionomys glareolus, Apodemus sylvaticus and white BK rats were given either a single intraperitoneal (i.p.) injection, an intragastric (i.g.) inoculation or an oral (p.o.) inoculation of the culture forms, including metacyclic trypomastigotes, of Trypanosoma microti, T. evotomys, T. grosi and T. lewisi, respectively. Similar levels of parasitaemia were produced by each of the three routes of infection, although the prepatent period was 3-5 days shorter in the case of the i.p.-injected animals. The oral inoculation of blood from mice infected with T. musculi into uninfected mice (outbred) and from rats infected with T. lewisi into uninfected BK rats produced parasitaemia after 6-8 days. This is the first report of the oral and i.g. transmission of T. microti, T. evotomys and T. grosi into their specific homologous hosts.

Differentiation of rodent trypanosomes of the subgenus Herpetosoma by lectins.

Trypomastigote forms of Trypanosoma microti, T. evotomys, T. grosi, T. musculi, and T. lewisi and trypomastigote and epimastigote forms of T. acomys were differentiated using 34 lectins and the Aminoff test for N-acetylneuraminic acid (NANA). Twelve of the lectins failed to agglutinate any of the above species. The number of lectins which agglutinated each species differed; T. mciroti, T. evotomys, T. grosi, T. musculi, T. lewisi and T. acomys (trypomastigote and epimastigote forms) were agglutinated by 7, 14, 7, 13, 11 and (11 and 10) lectins respectively. Some of the lectins were common in agglutinating all species of parasites, for example Bauhinia purpurea, Caragana aborescens and Viscum album. The minimum concentration of lectins which agglutinated each parasite was quite different. The agglutinations were cell body-cell body, flagellum-flagellum or flagellum-cell body. Most of the agglutinations were inhibited by their specific carbohydrates. The lowest concentrations of NANA was observed in T. lewisi (0.3 micrograms/ml) and the highest in T. musculi (4.5 micrograms/ml). The concentrations of NANA in T. microti, T. grosi and in T. acomys (trypomastigote and epimastigote forms) were 2, 1.8, 2.9, and (1.4 and 1.2) micrograms/ml respectively.

Studies on cross-immunity in Herpetosoma trypanosomes of Microtus, Clethrionomys and Apodemus.

Laboratory-bred rodents of three species were inoculated with heterologous Herpetosoma trypanosome species as follows: Microtus agrestis with Trypanosoma evotomys or T. grosi, Apodemus sylvaticus with T. evotomys or T. microti and Clethrionomys glareolus with T. grosi or T. microti. The three rodent species were subsequently challenged with their natural trypanosome parasite, i.e. T. microti for M. agrestis, T. grosi for A. sylvaticus and T. evotomys for C. glareolus. The parasitaemias and courses of infection that developed were followed. All challenged animals showed some degree of cross-immunity; not all became infected, and those that did had lower levels of parasitaemia and shorter patent periods than control animals. No C. glareolus previously inoculated with T. microti developed T. evotomys infections on challenge, and an infection was observed in just one of ten M. agrestis inoculated first with T. evotomys and later with T. microti.

In vitro cultivation of Herpetosoma trypanosomes on embryonic fibroblasts and in semidefined cell-free medium.

Microtus agrestis embryo fibroblasts (MAEF) support the survival and multiplication at 37 C of the mammalian multiplicative forms of the Herpetosoma trypanosomes Trypanosoma microti, T. evotomys, T. musculi, and T. lewisi passaged from cultures on Schneider's Drosophila medium and of T. grosi from Grace's medium. MAEF layers with parasites were maintained for a period of over 5 mo. A semidefined cell-free medium also supported the multiplication (at 37 C) of the mammalian forms of the same trypanosome species, passaged directly from Schneider's Drosophila medium or Grace's medium, without their prior culture on cell lines. Reproductive stages were observed in cultures; T. microti and T. evotomys produced nests of dividing amastigotes from which trypomastigotes developed in the medium supernatant. Trypanosoma lewisi, T. musculi, and T. grosi divided initially as epimastigotes, which then transformed to bloodstream trypomastigotes. Multiplication of trypomastigotes was also observed. These methods of reproduction are the same as those reported in the respective mammalian hosts.