Microscopic and Molecular Identification of Cyclospora cayetanensis and Cystoisospora belli in HIV-Infected People in Tabriz, Northwest of Iran.

Opportunistic pathogens such as Cryptosporidium, Cystoisospora belli, and Cyclospora cayetanensis cause various gastrointestinal and non-digestive disorders in people with HIV/AIDS. These symptoms are especially severe in HIV-infected people who have a CD4+ count of less than 200 cells/mL. This study aimed to determine the prevalence of C. belli and C. cayetanensis infections among people living with HIV in Tabriz, northwest of Iran. This descriptive study was performed on 137 people with HIV who had been referred to behavioral disease counseling centers in Tabriz. Then, after receiving written consent, fecal samples were collected and evaluated for the detection of parasitic infections using direct methods and modified acid fast staining, as well as polymerase chain reaction (PCR).From the 137 fecal samples collected (98 males and 39 females, between 20 and 40 years old), 1.5% were positive for C. cayetanensis and 2.9% were positive for C. belli. Due to the prevalence of C. cayetanensis and C. belli in people with HIV in Tabriz, essential measures, including personal hygiene training for infection control and prevention, seem necessary.

Molecular diagnosis of Trichomonas vaginalis in liquid-based Papanicolaou samples in Shiraz, southern Iran.

BACKGROUND: Trichomoniasis is a parasitic infection of the urinary and genital tract, caused by Trichomonas vaginalis. This study aimed to investigate the molecular diagnosis of T. vaginalis infection in liquid-based Papanicolaou samples in Shiraz, southern Iran. MATERIALS AND METHODS: In this cross-sectional study, 534 liquid-based Papanicolaou samples were collected from women referring to the laboratory of Motahari Clinic of Shiraz University of Medical Sciences in 2021. Genomic DNA were extracted from the samples and examined for evidence of T. vaginalis using polymerase chain reaction (PCR) using TVK3 and TVK7 specific primers. RESULTS: The mean age of participants was 39.28 ± 9.89 with a maximum age of 65 and a minimum age of 19 years. T. vaginalis DNA fragments were detected in 4.86% (26/534) of the cases. There was significantly higher prevalence in the age groups of 21 to 30 and 41 to 50 years (46.15%, p = 0.001 and 38.46%, p = 0.015, respectively). Furthermore, the results showed an association between a history of foamy discharge and Trichomonas positivity (p = 0.001). CONCLUSION: T. vaginalis infection is common in liquid-based Papanicolaou samples of women who attended regular health check-ups in the study area. Screening for trichomoniasis in populations, particularly if using highly sensitive methods such as PCR, may lead to increased detection and treatment.

Muscular hydatid cyst in Iran: A case report.

INTRODUCTION AND IMPORTANCE: Hydatid disease, caused by the larval stage of Echinococcus granulosus, is a common parasitic infection of humans and herbivores. Although livers and lungs are the most commonly affected organ, hydatid cysts may develop in any body part. Primary muscular hydatid cyst is extremely rare. CASE PRESENTATION: We reported the case of a 40-year-old-woman with the presentation of a soft, mobile, and non-tender lump in the dorsal part of her left upper arm (triceps brachii), which emerged one year ago. Her past medical history was unremarkable. The arm sonography revealed a single uniloculated cystic mass (6.5 cm × 5.5 cm) with a thick wall containing cystic lesions. It suggested the diagnosis of echinococcosis. The patient underwent surgery, and the hydatid cyst was excised. Histopathological examination confirmed hydatidosis. CLINICAL DISCUSSION: Hydatid cysts occur rarely (about 4 %) in muscles even in endemic regions. The study is the first case of hydatidosis found in triceps brachii in Fars province, Iran. In endemic regions, considering the hydatid cyst possibility is very important because it presents with many diversities. As it clinically presents a painless slow-growing mass, may be misdiagnosed with benign soft tissue tumors. CONCLUSION: Although muscular hydatidosis is extremely rare, it should be considered a differential diagnosis of any growing subcutaneous or muscular masses or tumors. Imaging modalities and blood tests are highly relevant for diagnosis. Surgical excision, a choice of treatment, should be done with cautions and is combined with anthelmintic therapy to reduce the risk of recurrence.

The Histopathological Findings in Appendectomy Specimens in an Iranian Population.

BACKGROUND: Appendicitis is one of the most common causes of acute abdominal surgeries. The importance of parasitic etiologies in the pathogenesis of appendicitis is not well known in appendectomy specimens on a large scale in southwestern Iran. The current study aimed to retrospectively assess the demographic data and histopathological records of appendicitis in a 28-year period in Fars province, southwestern Iran. MATERIALS AND METHODS: Histopathological records of 13,013 patients who had undergone surgeries for appendicitis at Dr. Ali Shariati Hospital, affiliated with the Fasa University of Medical Sciences from December 1993 to January 2021 were reviewed and data concerning the patients' demographic data and histopathological records were retrieved from each record. More than 6800 archived microscopic glass slides were also reviewed. RESULTS: From a total of 13,013 histopathological records of surgical excisions of appendicitis that were reviewed over a 28-year period, 8,189 (62.9%) were male and 4,842 (37.1%) were female. Patients' age ranged from 2 to 98 years, with a mean age of 24.68±19.87 years. The most common inflammatory changes were 5,687 (43.7%), 1,228 (9.4%), 670 (5.1%), 522 (4%), and 363 (2.8%) cases of acute appendicitis, suppurative appendicitis, early acute appendicitis, gangrenous appendicitis, and perforated appendicitis respectively. Microscopically, no viral inclusions, fungal elements, and histopathologic findings of bacterial causes were found. Parasitic infections such as helminthiasis were detected in 74 (0.6%) cases aged from 6 to 63. Enterobiasis (Syn. oxyuriasis, pinworm infection) accounted for 73 (98.6%) of the 74 helminthiases, while ascariasis accounted for 1 (1.4%). Out of 74 cases, 29 (39.2%) showed evidence of appendicitis. CONCLUSION: The results suggest that although parasitic agents are minor causes of appendicitis, these agents should be kept in mind during differential diagnosis. However, whether every parasitic infection leads to appendicitis is controversial.

Prevalence and Associated Risk Factors of Intestinal Parasitic Infections among Patients Visiting a Referral Hospital in Tehran Province, Iran.

Background: Intestinal parasitic infections (IPIs) are still considered a public health problem of mankind, particularly in immunocompromised patients. We aimed to determine the prevalence of IPIs with an emphasis on immunocompromised patients in a referral hospital in Tehran Province, Iran. Methods: In this cross-sectional study, 442 fecal specimens were collected randomly from patients, referred to Baqiyatallah Hospital in Tehran Province, Iran from May to September 2020. The collected specimens were examined using wet-mount, trichrome and modified Ziehl-Neelsen staining, formalin-ether concentration, and agar plate culture. Results: The prevalence of IPIs was found 17.0% (95% CI: 13.6-20.8%). The prevalence of protozoan parasites (16.3%; 95% CI: 13.0-20.1%) was significantly higher than helminthic parasites (0.7%; 95% CI: 0.1-2.0%). Blastocystis spp., Giardia lamblia, and Entamoeba coli were the most common intestinal protozoan with a prevalence of 12.2%, 1.6%, and 1.4%, respectively. E. histolytica/E. dispar/E. moshkovskii, Iodamoeba bütschlii, Cryptosporidium spp., Chilomastix mesnili as protozoan species and Hymenolepis nana, Dicrocoelium dendriticum, and Ascaris lumbricoides as helminthic species were the other detected parasites. Multiple logistic regression revealed a significant association of IPIs infections with stool consistency and the status of immune system. Conclusion: The prevalence of IPIs among the patients who are immunocompromised was significantly higher than immunocompetent patients (P< 0.05). Periodic stool examinations for screening of IPIs should be included as a part of routine medical check-up in these patients.

Promising Larvicidal Efficacy of Solid Lipid Nanoparticles Containing Mentha longifolia L., Mentha pulegium L., and Zataria multiflora Boiss. Essential Oils Against the Main Malaria Vector, Anopheles stephensi Liston.

PURPOSE: An attempt was made in the current study to develop a natural mosquito larvicide using nanotechnology. METHODS: Solid lipid nanoparticles (SLNs) containing three essential oils were first prepared using the high-pressure homogenizer. Larvicidal effects of essential oils and the SLNs against Anopheles stephensi were then compared. RESULTS: The size of SLN containing Mentha longifolia, Mentha pulegium, Zataria multiflora essential oil was obtained as 105 ± 7, 210 ± 4, and 137 ± 8 nm. Their zeta potentials were - 7.8, - 4.7, and - 9.7 mV. Besides, their efficacy with LC50 values of 24.79, 5.11, and 9.19 µg/mL was significantly more potent than that of their un-formulated essential oils with LC50 values of 36.2, 27.55, and 33.33 µg/mL. CONCLUSION: SLNs containing M. pulegium with the best efficacy (P < 0.05) could be considered as potent larvicides against other important species of mosquitoes and field trials.

A nanoemulsion-based nanogel of Citrus limon essential oil with leishmanicidal activity against Leishmania tropica and Leishmania major.

Cutaneous leishmaniasis is one of the diseases that severely affects human skin. Nanogels are the well-known formulation for topical drug delivery due to easy usage, high loading capacity, and physical and chemical stabilities. In this study, the toxicity effect of three essential oils, including Mentha piperita, Anethum graveolens, and Citrus limon (CLEO), was evaluated against Leishmania major and Leishmania tropica. Ingredients of CLEO as the most potent essential oil were identified using GC-MS analysis. The five major components were limonene (61.83%), sabinene (16.99%), trans-limonene oxide (3.08%), cis-limonene oxide (2.27%), and 1,2-cyclohexane diol, 1-methyl-4-(1-methylethenyl) (1.50%). The nanogel of CLEO (CLNgel) was prepared by the addition of carbomer 940 (1% w/v) to the prepared nanoemulsion with a droplet size of 146 ± 12 nm. The viscosity of CLNgel was fitted with a regression of non-Newtonian materials, Carreau-Yasuda. Interestingly, CLNgel at a concentration of 80 µg/mL reduced the viability of both species to 0%. Therefore, the prepared prototype can/could/would be used as an excellent nanoformulation for in vivo studies.

The present status of Sarcocystis spp. and sarcocystosis in Iran: a literature review.

Sarcocystosis due to Sarcocystis species, is prevalent among livestock in most parts of Iran, the predominant species being S. cruzi, and then followed by S. hirsuta and S. hominis. Studies on the prevalence and geographic distribution pattern of infection in the most common sites of infection have revealed infection rates up to 100% in at least one of the examined tissues in the country. Although human intestinal and gallbladder Sarcocystis infections have been reported, nothing is known about muscular Sarcocystis infection in human in Iran. The main aim of this review is to estimate the prevalence of Sarcocystis infections in ruminants, dogs, cats, and poultry as well as humans based on the studies conducted in Iran from November 1974 to October 2020. PubMed, Scopus, Web of Science, Science Direct, and Google Scholar, as well as one Persian electronic databases (SID) were searched systematically from November 1974 to October 2020. Publication searches were performed by various combinations of the following terms: "sarcocystosis" or "Sarcocystis spp." and "Iran". The reference list of selected articles was also manually screened, and the searching process resulted in the selection of 56 studies. The abstracts of the papers published at the congresses were not reviewed.

A survey of the frequency of cytomegalovirus-associated diarrhea in immunocompromised patients using a non-invasive method.

BACKGROUND AND OBJECTIVES: Cytomegalovirus (CMV) infection is the most common viral opportunistic infection causing gastrointestinal diseases such diarrhea and colitis in immunocompromised patients. The development and performance of a robust and sensitive PCR assay are usually evaluated to detect CMV DNA in human fecal specimens. In this study, our aim was to detect CMV DNA in stool samples taken from patients with HIV/AIDS, cancer, and transplant recipient patients with chronic and persistent diarrhea using a non-invasive method. MATERIALS AND METHODS: A total of 633 immunocompromised patients (451 males and 182 females) suffering from persistent or chronic diarrhea were included in this study. Among them, 392 were HIV/AIDS patients, 151 had cancer and were receiving chemotherapy, and 90 were recipients of a solid organ or bone marrow transplant. CMV genome was extracted from the stool samples using phenol: chloroform: isoamyl alcohol method. CMV DNA was identified by polymerase chain reaction using sequence specific primers on genomic DNA. RESULTS: Looking at the frequency of CMV DNA in 392 HIV/AIDS patients, we found that only 5 patients (1.27%) were positive for CMV genome, while this frequency was 4.63% (7/151) and 5.5% (5/90) in patients with cancer receiving chemotherapy and in those with solid organ or bone marrow transplant, respectively. CONCLUSION: The results of this study revealed that the cause of chronic or persistent diarrhea in HIV/AIDS, cancer, and graft recipient patients might be related to CMV infection. Accordingly, we recommend a non-invasive method, such as stool sample, as a first line of diagnosis of enteritis when the physician suspects that a patient has CMV infection.

Human intestinal sarcocystosis in Iran: there but not seen.

Human sarcocystosis is a rare infection caused by the genus Sarcocystis who human serve as definitive (intestinal form of infection) host or intermediate (extraintestinal form) host for some various Sarcocystis species. The detection of Sarcocystis oocysts/sporocysts in the feces usually incidentally and is achieved by microscopic examination of the fresh or preserved specimens. To rule out any parasitological etiology among 23,875 (aged 2 months to 95 years) apparently immunocompetent Iranian individuals (from October of 2010 to June of 2016) with abdominal discomforts referred to several teaching hospitals and local clinical laboratories in Fars Province, Iran, their fecal samples were examined using light microscopy. Most pathogenic parasite-positive and doubtful samples were sent to the Intestinal Protozoology Laboratories of Fasa and Shiraz Universities of Medical Sciences to further examination to detect probable co-infection with other underdiagnose parasitoses. In addition to the common protozoal and helminthic infections, during the course of examining stool specimens using direct smear mixed with saline or iodine mounts and by formalin-ethyl acetate techniques, four cases of intestinal Sarcocystis infection as only or concurrently infected with other parasites were found. The present paper presents cases of human intestinal Sarcocystis infection in Iran. Since Sarcocystis are small in size and usually rare in stool, they often go unnoticed. It should be noted that stool smears must be examined with great care to avoid misinterpretation of Sarcocystis infections in microscopic examinations. To the best of our knowledge, co-infection of intestinal sarcocystosis and other principal parasitoses in stool investigations has not been reported earlier.

Prevalence of Enteric Protozoan Oocysts with Special Reference to Sarcocystis cruzi among Fecal Samples of Diarrheic Immunodeficient Patients in Iran.

The genus Sarcocystis is not usually considered as an important enteric pathogen in immune compromised patients. It might be expected that species for which humans are the final host (Sarcocystis hominis and Sarcocystis suihominis as well as possibly others) would be encountered increasingly often in immunodeficient persons. This study aimed to address how to detect and differentiate Sarcocystis oocysts and/or sporocysts from enteric protozoans in the diarrheal samples of immunodeficient patients in Shiraz, Iran. Diarrheal samples of 741 immunodeficient patients with recurrent persistent or chronic diarrhea were examined by microscopy and molecular biological analysis. Oocysts-positive samples were 68 Cryptosporidium spp., 9 Cystoisospora belli (syn. Isospora belli), 2 Cyclospora cayetanensis, and 15 microsporidia (Enterocytozoon bieneusi). Sarcocystis-like sporocysts found from a woman were identified as Sarcocystis cruzi through 18S rDNA amplification and phylogenetic analysis. To the best of our knowledge, this is the first report of S. cruzi from a human.

Cystoisosporiasis-related human acalculous cholecystitis: the need for increased awareness.

Cholecystitis is one of the common surgical indications affecting human beings in many countries. A variety of infectious agents can be associated with acute or chronic acalculous cholecystitis, especially in HIV/AIDS patients. In this investigation, the authors aim to describe two cases of histologically and molecularly documented cystoisosporiasis (syn. isosporiasis) as the cause of chronic acalculous cholecystitis in two immunodeficient patients. During microscopic examinations of more than 2500 diarrheic patients' samples, 11 cases of cystoisosporiasis-related recurrent persistent/chronic diarrhea were detected. A review of the medical records of Cystoisospora belli (syn. Isospora belli)-positive patients showed that two of them, i.e. a patient with prolonged corticosteroid therapy and an AIDS patient, several months prior to fecal examinations had undergone cholecystectomy due to acalculous cholecystitis. The study was continued by a review of the histopathological investigation of the recuts prepared from the excised gallbladder tissue sections and stained with hematoxylin and eosin in order to detect a possible specific clinical correlation with cystoisosporiasis. Light microscopic examination revealed the presence of various developmental stages of a coccidial parasite, namely Cystoisospora belli, in both patients' gallbladder tissue sections. To the best of our knowledge, C. bellii-associated cholecystitis has not been previously reported in a patient with prolonged corticosteroid therapy.

First detection of acalculous cholecystitis associated with Sarcocystis infection in a patient with AIDS.

Acalculous cholecystitis and cholangitis are increasingly being recognized as complications of AIDS. The opportunistic parasites that have been most commonly associated with these disorders are Cryptosporidium species, Isospora belli, Cyclospora cayetanensis and Enterocytozoon bieneusi. The authors performed a parasitological survey on the gallbladder tissue sections of patients underwent cholecystectomy due to chronic acalculous cholecystitis at the Shiraz University of Medical Sciences, Iran. Light microscopic investigation in more than three hundred archived histopathological slides revealed the presence of sexual stages (i.e., mature sporocysts) of a coccidial protozoan in a patient with AIDS who developed acalculous cholecystitis as confirmed by histological, parasitological and molecular tests in which Sarcocystis species was the only identifiable pathogen in gallbladder sections. In the best of our knowledge it's the first documented case of chronic non-calculous cholecystitis due to Sarcocystis parasite in an Iranian AIDS patient from worldwide.

Microsporidia and coccidia as causes of persistence diarrhea among liver transplant children: incidence rate and species/genotypes.

We determined species/genotype(s) of enteric microsporidia and coccidia causing diarrhea among 44 liver transplant children in Shiraz Nemazee hospital using acid-fast-trichrome staining and polymerase chain reaction-sequencing techniques. Enterocytozoon bieneusi (genotype D), Cryptosporidium (parvum and meleagridis) were detected in 6.81% and 11.36% of the children, respectively.

HIV/AIDS-associated opportunistic protozoal diarrhea.

Human immunodeficiency virus (HIV) infection has altered both the epidemiology and outcome of enteric opportunistic parasitic infections. This study was done to determine the prevalence and species/genotypes of intestinal coccidian and microsporidial infections among HIV/AIDS patients with diarrhea and/or a history of diarrhea alternately with an asymptomatic interval, and their association with CD4 T cell count. This cross-sectional study was done from May 2010 to May 2011 in Shiraz University of Medical Sciences, South of Iran. A blood sample was obtained from HIV-positive patients for a CD4 T cell count upon enrollment. Sociodemographic data and a history of diarrhea were collected by interviewing 356 consecutive participants (273 males and 83 females). Whenever possible more than a fecal sample was collected from all the participants and examined for parasites using direct, physiological saline solution ethyl acetate, an acid-fast trichrome stain, nested polymerase chain reaction, and sequencing techniques for the detection, confirmation, and genotyping of Cryptosporidium spp., Cyclospora cayetanensis, Isospora belli, and intestinal microsporidia (Enterocytozoon bieneusi). The most common opportunistic and nonopportunistic pathogens were Cryptosporidium spp. (C. parvum and C. andersoni), E. bieneusi, Giardia lamblia, Sarcocystis spp., and Blastocystis homonis affecting 34, 8, 23, 1, and 14 patients, respectively. C. cayetanensis, I. belli, Enterobius vermicularis, and Hymenolepis nana were observed in few patients. A CD4 count <200 cells/µl was significantly associated with the presence of opportunistic parasites and diarrhea (p<0.05). Opportunistic intestinal parasites should be suspected in any HIV/AIDS patient with chronic diarrhea. Tropical epidemic nonopportunistic enteric parasitic infections among such patients should not be neglected in Iran.