Candida
species cause a wide range of opportunistic infections in humans and animals. The detection of
Candida
species by conventional diagnosis methods is costly and time consuming. This study was ...conducted for the first time to evaluate and compare a relatively new molecular assay and the loop-mediated isothermal amplification (LAMP) technique with conventional methods for detection of
Candida albicans
. In this study, 70 different species of
Candida
identified by conventional methods were cultured on Sabouraud chloramphenicol agar medium and then the genomic DNA was extracted. The LAMP technique was performed using specific primers targeting the ITS2 gene of
C. albicans
. The analytical sensitivity and specificity of LAMP were measured using a tenfold serial dilution prepared from extracted DNA from standard
C. albicans
strain from 1 ng to 1 fg and the DNA samples of other clinical
Candida
species and three non-
Candida
yeast. Out of 70 yeast samples analyzed by LAMP technique, 24 samples (34.3%) were positive for
C. albicans
. Comparison of the results showed that the CHROMagar
Candida
and germ tube production methods are quite consistent with the LAMP technique, while the agreement amount between the results of carbohydrate assimilation and chlamydoconidia generation assays and LAMP technique was 98.5% and 72.8%, respectively. The detection limits of the LAMP assay were 10 fg of the DNA from the standard
C. albicans
strain. No amplification was observed in the DNA samples of other yeast species and only the DNA sample of standard
C. albicans
strain was amplified. Based on the results, it can be concluded that the LAMP method is as specific and precise as common diagnostic methods, but is faster, easier deployable or more sensitive. Therefore, this method can be used as a suitable complementary assay for
Candida
diagnosis in medical diagnostic laboratories and field conditions.
Human infection with Enterobius vermicularis occurs worldwide, particularly in children. The role of E. vermicularis in appendicitis is neglected. This study was designed to investigate genotypes of ...E. vermicularis detected from appendectomy specimens in the human population from Iran and clarify the intra-species variation of the parasite. Seventy appendectomies for acute clinical appendicitis isolates from Azerbaijan and North Khorasan of Iran were used in the present study. The genetic information of Tehran and Hamedan regions was also obtained from GenBank for comparison and analysis. The nucleotide sequence of cytochrome c oxidase subunit 1 (cox1) gene was analyzed to perform genetic differentiation, haplotype network analysis, and population structure. Phylogenetic analysis of all the isolates were included in type B haplogroup. The number of haplotypes in all geographical locations of Iran is not much. Network analysis of sequences for regions such as Thailand, Iran, Denmark, and Poland show three classified subtypes B1, B2, and B3 in the B haplogroup. It seems that the haplotypes of E. vermicularis detected from appendectomy are B type, and divided into three subtypes. Further research using another genetic marker is required to elucidate the genetic variation of the parasites in detail.
Background. Human toxocariasis (HT) is a zoonotic disease with a global expansion. Contaminated soil with Toxocara spp. eggs is the main source of human infection, which may lead to severe ...complications depending on the organs invaded by migrating larvae. Aim. This study is aimed at eliciting the prevalence of Toxocara spp. eggs in public parks in Zahedan, southeast Iran, and providing new insight into the soil contamination rate in this area using microscopic and molecular methods. Methods. Based on five municipal districts, 240 soil samples were collected from public parks and playgrounds in Zahedan. The modified Sheather’s flotation technique was employed to isolate Toxocara spp. eggs from the soil, followed by microscopic assessment and molecular evaluation of internal transcribed spacer 1 and 2 ribosomal deoxyribonucleic acid (ITS1 and 2 rDNA) using nested polymerase chain reaction (nested PCR) to identify the presence of Toxocara spp. eggs. The Sanger sequence was used to differentiate the Toxocara species. Subsequently, all the sequenced data were blasted and compared with other sequences available in the GenBank. Results. Out of 240 soil samples collected, 7 (2.9%) samples were identified to contain Toxocara spp. eggs using Sheather’s flotation and microscopic techniques. Meanwhile, 19 (7.9%) samples were positive using nested PCR. According to the Sanger sequencing analysis findings, all positive samples were contaminated with Toxocara cati. Conclusion. As evidenced by the obtained results, only T. cati species were detected in public parks and playgrounds in Zahedan; therefore, control and prevention programs against this species should be considered in human and animal communities.
One of the opportunistic pathogens which cause serious problems in the human immune system is Toxoplasma gondii, with toxoplasma encephalitis (TE) seen in patients affected by it. The treatment of ...these patients is limited, and if not treated on time, death will be possible.
In this study, the effects of the treatment with different doses of fluconazole (FLZ) in combination with the current treatment of acute toxoplasmosis on reducing the mortality rate and the parasitic load in the murine model in vivo were studied. The mice were treated with different doses of fluconazole alone, sulfadiazine, and pyrimethamine plus fluconazole. A day after the end of the treatment and 1 day before death, the mice's brains were collected, and after DNA extraction and molecular tests, the parasite burden was detected.
This study showed that a 10-day treatment with 20 mg/kg of fluconazole combined with sulfadiazine and pyrimethamine 1.40 mg/kg per day affected acute toxoplasmosis and reduced the parasitic load significantly in brain tissues and also increased the survival rate of all mice in this group until the last day of the study, in contrast to other treatment groups. These results also indicate the positive effects of combined therapy on Toxoplasma gondii and the prevention of relapse.
Reducing the parasitic burden and increasing the survival rate were more effective against acute toxoplasmosis in the combined treatment of different doses of fluconazole with current treatments than current treatments without fluconazole. In other words, combination therapy with fluconazole plus pyrimethamine reduced the parasitic burden in the brain significantly, so it could be a replacement therapy in patients with intolerance sulfadiazine.
Background & Aims: Blastocystis sp. is a common parasite of humans with a vast variety of nonhuman hosts and global distribution. Based on molecular methods, distinct subtypes are isolated from ...different hosts. In this study, we detected human-isolated Blastocystis subtypes in the northwestern of Iran. Materials & Methods: A total of 600 stool samples referred to health centers in Urmia and Maragheh cities in the northwest of Iran was randomly collected and examined using direct wet mount, formalin-ether concentration, a modified version of the Ziehl–Neelsen staining technique for the detection of parasitological items and using barcoding method (18S rRNA gene) for the molecular survey. Results: From 600 samples studied, 496 (82.7%) and 104 (17.3%) were male and female, respectively. Three subtypes, including ST1, ST2 and ST3, were identified from collected samples, and the frequent subtypes were ST3, ST1, and ST2, respectively. Conclusion: While genetic similarity of Blastocystis isolated from human and animal did not show in this region, the zoonotic cycle of this parasite, probably by fecal contamination, exists in the study area. By identifying Blastocystis subtypes, it is possible to prevent the transmission and related complications.
Neonatal hypoxia-ischemia (HI) is one of the most important causes of neurological disorders in children. Various studies suggest that maternal exercise during pregnancy has a beneficial impact on ...the health status of offspring infants. In this study, the effect of maternal treadmill exercise during pregnancy on neurological and molecular changes induced by HI in newborn rats was investigated.
In this experiment, 24 pregnant female rats were divided into two groups; the first group was subjected to treadmill exercise for six weeks. The treadmill exercise program was initiated by running for 17 min at 5-10 m/min at 0 inclination in the first week, followed by running for 21 min at 5-25 m/min at 5° inclination in the second week, running for 25 min at 5-30 m/min at 10° inclination in the third and fourth weeks, running for 25 min at 5-15 m/min at 10° inclination in the fifth and sixth weeks. The second group was left untreated and did not perform the exercise. Newborn rats were assigned to four groups; (1) control, (2) control+exercise, (3) HI, and (4) HI+exercise. HI was developed in the offspring on the 8th postnatal day. One week following the induction of HI, the Garcia test was carried out. The histological morphology of neonates was assessed, and the expression levels of caspase-1 and NLRP3 were evaluated.
The data showed that maternal exercise during pregnancy significantly improved neural cell death (
<0.001) and the Garcia score (
<0.05), while it attenuated the expression levels of caspase-1 (
<0.001) and NLRP3 (
<0.05) genes in newborn rats induced by HI.
These results demonstrated that maternal treadmill exercise during pregnancy could reverse the neurological deficits, as well as the expression levels of caspase-1 and NLRP3 genes, which occur in neonatal hypoxia-ischemia.
Abstract This report describes a rare case of ophthalmic dirofilariasis in a 2-year-old boy with redness, irritation, pain and foreign body sensation in the right eye. Slit lamp examination ...demonstrated a thread-like whitish nematode in the anterior chamber of the right eye that twisted around itself. The nematode worm (35 mm long and 150–200 μm width) was removed surgically. The presence of the smooth cuticular surface without longitudinal ridges and the vulva showed that it could be a female Diroflaria immitis . PCR amplification was done to verify the Diroflaria species. PCR amplification and sequence analysis of mitochondrial 12S rDNA confirmed that recovered worm was D. immitis . Ocular dirofilariasis caused by D. immitis is very rare, but it must be considered in humans living in endemic areas.
Babesiosis is a globally distributed zoonotic parasitic disease in a broad range of vertebrates with great importance in the veterinary field. The standard diagnostic test for Babesiosis in animals ...is microscopic identification of the parasite in a venous blood smear stained with Giemsa combined with assessment of clinical manifestations throughout the acute phase of the disease. The present study was planned to determine the presence of Babesia species in camels from the southeastern regions of Iran. A total of 140 blood samples of camels were randomly collected in four selected cities including Qaen, Nehbandan, Iranshahr, and Zahedan from March to August 2019. Blood smears of each case were also examined by the Giemsa staining method and extracted DNA samples were subjected to internal transcribed spacers (ITS1) polymerase chain reaction (PCR) amplification. The prevalence rates using microscopically and molecular examinations were 10% and 19.28%, respectively. The prevalence rates significantly vary between the selected regions (p = 0.003). PCR technique showed higher sensitivity than microscopy. We found that all infected camels were positive for Babesia caballi. The rate of infection with Babesia among the camel in Zahedan is remarkable. Early diagnosis and early treatment can prevent further spread of the disease in this area.
Babesisosis in camels can be detected by microscopic and molecular diagnosis throughout the acute phase of disease. In the present study, Babesia species in camels from the eastern regions of the Iran was determined.
Background. The purpose of present study was to assess the impact of maternal treadmill exercise during pregnancy on inflammation, oxidative stress, expression of Bax and Bcl-2 genes, and ...brain-derived neurotrophic factor (BDNF) level in neonatal rat brain after the hypoxia-ischemia injury. Material and Methods. A total of 24 female Wistar rats were utilized in this research. Two groups are randomly considered for rats: (1) not exercised through pregnancy and (2) exercised during pregnancy. Offsprings were divided into four groups including after delivery: (1) sham, (2) sham/exercise (sham/EX), (3) HI, and (4) HI+exercise. HI was induced in pups at postnatal day 8. Neurobehavioral tests were done seven days after HI induction. Then, the brain tissue was taken from the skull to estimate Bcl-2 and Bax gene expressions, BDNF, cerebral edema, infarct volume, inflammatory factors, oxidative stress, and neurological function. Results. The BDNF level in the HI+exercise group was considerably higher than the HI, sham, and sham/EX groups. Tumor necrosis factor (TNF-α), C-reactive protein (CRP), and the whole oxidant capacity (TOC) levels in the HI group were significantly higher than the sham and sham/EX groups. TNF-α, CRP, and TOC levels in the HI+exercise group were significantly lower than the HI group. Total antioxidant capacity (TAC) level in the HI+exercise group was significantly higher than the HI group. Infarct volume and edema percent in the HI+exercise group were significantly lower than the HI group. Neurological function in the HI+exercise group was significantly better than the HI group. Bax expression in the HI+exercise group was significantly lower than the HI group. Bcl-2 expression in the HI+exercise group was significantly higher than the HI group. In the sham group, BDNF, TNF-α, CRP, TAC, TOC, edema levels, and neurological function had no significant difference with the sham/EX group. Conclusion. It appears that the maternal treadmill exercise during pregnancy exerts a supportive impact against neonatal HI brain injury through increasing antioxidant capacity, Bcl-2 expression, and BDNF levels and decreasing inflammation that is resulted in the lower infarct volume and sensorimotor dysfunction.
Objective(s): Attempts to produce vaccines for leishmaniasis need adjuvants to trigger the kind of immune reaction required for protection. In this study, we examined the properties of the TLR7 ...agonist imiquimod, a vaccine adjuvant, making use of a live model of infection where the immune reactions could be identified prior to and following the challenge of infection.
Materials and Methods: The liposomes of EPC containing the imiquimod adjuvant were prepared and characterized for protein concentration, surface charge, and particle size. Vaccination was done using the soluble Leishmania antigen (SLA) as a first-generation vaccine model in the liposomal state to vaccinate BALB/c mice against the challenge of leishmania major. BALB/c mice were vaccinated subcutaneously, three times at a two-week interval. Parasite burden, footpad swelling, IgG isotype, as well as the level of IL-4 and IFN-γ were assessed as the protection criteria.
Results: The group of mice vaccinated by Lip+Imiquimod+SLA demonstrated a lower amount of footpad swelling and parasite burden than the buffer group. In addition, the highest level of IFN-γ and the lowest level of IL-4 production was noticed in the splenocytes of the mice vaccinated with the formulation of Lip+Imiquimod+SLA.
Conclusion: These results imply that imiquimod added to the formulation of liposomes is able to modulate the immune reaction of the BALB/c mice vaccinated preferably to a Th1 reaction rather than a Th2 reaction which can also lead to partial protection against the challenge of Leishmania.
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