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BACKGROUND: The sensitivity of parasitological and molecular methods is unsatisfactory for the diagnosis of strongyloidiasis, and serological techniques are remaining as the most effective diagnostic approach. The present study aimed to design and produce a chimeric recombinant antigen from Strongyloides stercoralis immunoreactive antigen (SsIR) and Ss1a antigens, using immune-informatics approaches, and evaluated its diagnostic performance in an ELISA system for the diagnosis of human strongyloidiasis. METHODOLOGY/PRINCIPAL FINDINGS: The coding sequences for SsIR and Ss1a were selected from GenBank and were gene-optimized. Using bioinformatics analysis, the regions with the highest antigenicity that did not overlap with other parasite antigens were selected. The chimeric recombinant antigen SsIR- Ss1a, was constructed. The solubility and physicochemical properties of the designed construct were analyzed and its tertiary structures were built and evaluated. The construct was expressed into the pET-23a (+) expression vector and the optimized DNA sequences of SsIR-Ss1a (873 bp) were cloned into competent E. coli DH5α cells. Diagnostic performances of the produced recombinant antigen, along with a commercial kit were evaluated in an indirect ELISA system, using a panel of sera from strongyloidiasis patients and controls. The physicochemical and bioinformatics evaluations revealed that the designed chimeric construct is soluble, has a molecular with of 35 KDa, and is antigenic. Western blotting confirmed the immunoreactivity of the produced chimeric recombinant antigen with the sera of strongyloidiasis patients. The sensitivity and specificity of the indirect ELISA system, using the produced SsIR-Ss1a chimeric antigen, were found to be 93.94% (95% CI, 0.803 to 0.989) and 97.22% (95% CI, 0.921 to 0.992) respectively. CONCLUSIONS/SIGNIFICANCE: The preliminary findings of this study suggest that the produced SsIR-Ss1a chimeric antigen shows promise in the diagnosis of human strongyloidiasis. However, these results are based on a limited panel of samples, and further research with a larger sample size is necessary to confirm its accuracy. The construct has potential as an antigen in the ELISA system for the serological diagnosis of this neglected parasitic infection, but additional validation is required.
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Antígenos de Helmintos , Ensaio de Imunoadsorção Enzimática , Sensibilidade e Especificidade , Testes Sorológicos , Strongyloides stercoralis , Estrongiloidíase , Humanos , Estrongiloidíase/diagnóstico , Estrongiloidíase/imunologia , Animais , Testes Sorológicos/métodos , Ensaio de Imunoadsorção Enzimática/métodos , Strongyloides stercoralis/imunologia , Strongyloides stercoralis/genética , Antígenos de Helmintos/genética , Antígenos de Helmintos/imunologia , Anticorpos Anti-Helmínticos/sangue , Proteínas Recombinantes de Fusão/imunologia , Proteínas Recombinantes de Fusão/genética , Escherichia coli/genética , Escherichia coli/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Expressão GênicaRESUMO
Background: Toxocara canis is one of the most important causes of animal toxocariasis with global distribution. We aimed to find out the seroprevalence of toxocariasis in dogs in a rural area in Fars Province, south of Iran. Methods: Dogs blood samples were collected from 60 dogs in three rural areas in the Sar Mashhad region, Fars Province. Dog sera were evaluated for anti-Toxocara antibodies by an indirect ELISA method. The association between the seropositivity and age, gender, and the sampling location were statistically evaluated. Results: Serological assay detected anti-Toxocara antibodies in sera of 32 out of 60 dogs, corresponding to a seroprevalence of 53.3%. The rate of seropositivity was higher in the male dogs. The rate of seropositivity was higher in old dogs. This rate increased with increasing age, however, the association between age and Toxocara seropositivity was not statistically significant. Conclusion: The high prevalence of Toxocara infection in dogs in the current study area confirms that infected dogs are an important source of Toxocara infection for their owners and people who are in close contact with these animals, especially children.
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Cutaneous leishmaniasis is of particular importance in southern Iran. This study aimed to investigate the infection of rodents with Leishmania major in an urban area of Fars Province, located in southern Iran. Rodents were trapped and samples from the liver, spleen, and skin were collected. Impression smears were prepared from these tissues and any skin lesions and were examined microscopically. In addition, a portion of the samples were preserved for subsequent DNA extraction. A total of 41 rodents belonging to three species were caught from 10 trapping stations in gardens or houses within the area. The caught rodent species were Rattus rattus (n = 25, 60.97%), Mus musculus (n = 15, 36.58%), and Meriones persicus (n = 1, 2.5%). Leishmania amastigotes were seen in the spleen tissue smear of 6 (2.43%) of the rodents, including 4 of R. rattus and 2 of M. musculus. Skin lesions were observed on the muzzles of two R. rattus and one M. musculus. Samples taken from these lesions tested positive for Leishmania infection. Leishmania DNA was detected in 18 (43.9%) rodents, including 11 R. rattus, 6 M. musculus, and one M. persicus, based on DNA sequencing of the ITS2 gene and PCR of the kDNA. Phylogenetic reconstruction revealed that the parasite infecting the rodents was L. major. The detection of Leishmania infection in these rodents in urban areas raises concerns about the urbanization of cutaneous leishmaniasis caused by L. major. This urbanization poses unique challenges for control and prevention efforts.
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Background: Inflammation and oxidative activities within the gut play major roles in the pathogenesis of ulcerative colitis (UC). We aimed to determine the effect of Melissa officinalis, an antioxidant and anti-inflammatory agent, on the colon histological characteristics in acetic acid (AA)-induced UC in rat models. Methods: Thirty-six male rats with AA-induced colitis were divided into 5 groups: no treatment (AA); daily treatment with 300 mg/kg Melissa officinalis orally (MO) and rectally (MR); and 100 mg/kg mesalamine orally (AO) and rectally (AR). Macroscopic and histopathological evaluation of the colon, along with a biochemical laboratory evaluation, were performed 10 days after UC induction. Results: All treatment groups demonstrated lower macroscopic grading scores compared to the AA group. After treatment with MO, 42.9% of cases demonstrated no macroscopic changes, while 14.3% demonstrated only mucosal erythema. In the MR group 28.6% of rats had no changes in their mucosal lining and 28.6% had only mucosal erythema. Following histopathological evaluation, the AO group had lower scores regarding the severity of ulcer, inflammation, destruction, crypt abscess, and disorganization compared to the MO group. (P=0.02) The MR group demonstrated lower microscopic scores compared to the MO group, and also lower macroscopic scores compared to the AR group, although not significantly (P>0.05). Conclusions: Both oral and topical administration of Melissa officinalis have satisfactory healing properties compared to mesalamine, with topical route having better results. Therefore, further studies are needed to establish the benefit of Melissa officinalis administration (both orally and topically) within a UC treatment protocol.
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OBJECTIVE: Superinfection of cystic echinococcosis (CE) is a life-threatening complication with significant morbidities, which can be prevented with early diagnosis and treatment. This study aims to examine the clinical characteristics, diagnostic methods, and treatment options for superinfected CE, as there is currently limited information available on the differences between superinfected and non-infected CE in terms of clinical features, serological and radiologic findings. METHODS: This cross-sectional study was conducted on hospital records of patients who were diagnosed with hydatid cysts in a 15-year period (2004 to 2018) in two main university-affiliated referral centers in Fars province, southern Iran. Patients' information regarding the demographical and clinical features related to CE, age, sex, previous history of CE or recurrence, size and location of CE, and length of hospital stay were collected. Moreover, the characteristics of concurrent infections with hydatid cysts were recorded. RESULTS: A total of 586 surgeries due to CE were performed on 501 patients, of which 67 (11.43%) had reoperations due to the recurrence of the disease. A total of 30 (5.99%) incidences of superinfection were observed. There were no statistically significant differences in terms of laboratory and imaging findings between CE patients with concurrent infections and other CE patients (p-value > 0.05). Among the patients with super-infection, four had fungal infections of the lungs. Aspergillus fumigatus was the causative pathogen in all four patients that were diagnosed with fungal superinfection. All patients underwent surgical excision with favorable long-term outcomes. CONCLUSION: Our study revealed a 5.99% incident rate of CE superinfection. Regarding the concurrent fungal infections in hydatid cysts, the patient's symptoms and laboratory and imaging findings are inconclusive and histopathological evaluation seems to be the most reliable option. Surgical resection is the gold-standard treatment option with favorable outcomes and potentially can be curative.
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Equinococose , Echinococcus , Micoses , Superinfecção , Animais , Humanos , Estudos Retrospectivos , Irã (Geográfico)/epidemiologia , Superinfecção/epidemiologia , Estudos Transversais , Equinococose/complicações , Equinococose/epidemiologia , Equinococose/diagnósticoRESUMO
Background: We aimed to present a systematic review and meta-analysis of studies that used antigen-based assays for the diagnosis of human giardiasis. Methods: All the related published literature cited within PubMed, ISI web of science, Google Scholar, Embase, and Scopus, were searched up to December 2021. The search terms, both as MeSH terms and text words, were "Giardia", "Giardia lamblia", "Giardia intestinalis", "giardiasis", combined with "diagnosis", "antigen detection", serodiagnosis, or serological diagnosis. The required data was extracted from the papers. Pooled estimates of sensitivity and specificity were obtained and forest plots and summary receiver operating characteristics (SROC) plots were used to calculate sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV). Results: The search of databases found 1683 papers, of which 46 articles fulfilled our eligibility criteria. The sensitivity of antigen-based methods for the diagnosis of human giardiasis ranged from 45% (95% CI: 31-59%) to 100% (95% CI: 100-100%) and the pooled estimate of sensitivity was 92% (95% CI: 90-93%). The pooled estimated specificity was 97% (95% CI: 96-98%), ranged from 81% (95% CI: 68-89%) to 100% (95% CI: 98-100%). The summary estimate of PPV and NPV were 92 % (95% CI: 90-93%) and 97% (95% CI: 96-98%) respectively. Comparing the performance of the antigen detection assays by region revealed a significant difference in the assay's performances in different regions of the world. Conclusion: The antigen-based detection methods have acceptable and satisfactory performance in the diagnosis of human giardiasis. The task ahead is to identify more specific target antigens and design simpler, cheaper, and more sensitive methods for the diagnosis of this common worldwide-distributed parasitic infection.
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BACKGROUND: The lungs are considered the second-most frequent location for hydatid cyst in human. The current retrospective hospital-based study aimed to assess the epidemiological data, clinical presentation, and treatment outcomes of lung hydatid cyst in patients who underwent surgery for this disease in Fars province, southern Iran. METHODS: In this retrospective study, hospital records of 224 pulmonary hydatid cyst patients were assessed in two main university-affiliated hospitals in Fars Province, southern Iran. Clinical features of patients, epidemiological data, cyst features, surgical interventions, and treatment outcomes were reviewed and analyzed. RESULTS: A total of 224 hydatid cyst cases of the lung were reviewed. Male patients accounted for the majority of cases (60.4%). The average age of the patients was 31.13 (± 19.6), ranging from 2 to 94 years old. Of the 224 patients, 145 (75.9%) cases had only one single cyst and mostly 110 (53.9%) located in the right lung. Also, 6 (2.9%) cases had cysts in both lungs. The lower lobe of the lungs was the most common location of the hydatid cyst. The average size of lung hydatid cyst was 7.37 cm (SD = 3.86; rang: 2-24) while for the cyst areas was 42.87cm2 (SD = 52.76; range: 2-488). Regarding the surgical method, 86 (38.6%) cases were operated by lung resection surgery while 137 (61.4%) cases had lung preserving one. The chief complaints of the patients were cough (55.4%) and dyspnea (32.6%). Relapse was documented in 25 (11.16%) of cases. CONCLUSIONS: Lung hydatid cyst is a common infection in southern Iran. Lung preserving surgery is the method of choice for the management of hydatid cyst. Relapse, which was not uncommon in our study, is a challenging feature of hydatid cyst management.
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Cistos , Equinococose Pulmonar , Humanos , Masculino , Pré-Escolar , Criança , Adolescente , Adulto Jovem , Adulto , Pessoa de Meia-Idade , Idoso , Idoso de 80 Anos ou mais , Equinococose Pulmonar/cirurgia , Estudos Retrospectivos , Irã (Geográfico)/epidemiologia , Recidiva Local de Neoplasia , Pulmão , Hospitais UniversitáriosRESUMO
Background: Toxocariasis is an important zoonotic infection, especially in tropical areas. One of the significant challenges in the serodiagnosis of human toxocariasis is the cross-reaction of Toxocara antigens with other parasites due to their relatively similar glycan structures. Removing the glycan structure from Toxocara excretory-secretory (TES) antigens may increase the efficacy of these antigens in the diagnosis of toxocariasis. The current study aimed to assess the efficacy of deglycosylated Toxocara cati excretory-secretory (dTES) antigens for the serodiagnosis of human toxocariasis. Methods: Toxocara ES antigens were prepared from T. cati second-stage larvae and deglycosylated using sodium hydroxide (NaOH). The TES antigens, along with the dTES antigens, were used in an ELISA as well as a western blotting system for the detection of anti-Toxocara antibodies. Sera samples collected from 30 confirmed cases of toxocariasis, 30 patients with other diseases, and 30 healthy subjects were evaluated by both systems. Results: The sensitivity of TES and dTES ELISA for the diagnosis of human toxocariasis was 96.67% (95% CI = 82.78-99.92) and 93.33% (95% CI = 77.93-99.18), respectively, while the specificity of dTES (88.33%; 95% CI = 77.43-95.18) increased significantly compared to the TES (80.00%; 95% CI = 67.67-89.22). The sensitivity of both antigens was 100% (95% CI = 88.43-100) by the western blotting system. Moreover, the specificity of TES and dTES antigens was 95% (95% CI = 86.08-98.96) and 98.33% (95% CI = 91.06-99.96), respectively, when using the western blotting system. Conclusion: Results of the current study indicate that the chemical removal of the glycan epitopes of T. cati ES antigens significantly reduces cross-reactivity rates with other parasitic infections. Considering the findings of the present study, the dTES antigens seem to be suitable antigens for the serodiagnosis of human toxocariasis.
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Background: The current study aimed to evaluate the therapeutic features and complications of liver hydatid cyst in patients who underwent surgery for cystic echinococcosis (CE) in Fars province, southern Iran. Methods: A total of 293 patients who underwent surgery for liver hydatid cyst from 2004 to 2018 in Fars province, southern Iran, were retrospectively evaluated. The clinical records of patients were reviewed, and the demographic and clinical characteristics of each patient were assessed. Results: Of the total of 293 cases, 178 (60.9%) were females and 115 (39.1%) were males. The mean age of the subjects was 37.22 (±20.55) years. The mean size of the liver hydatid cyst was 9.18 (±4.365) cm. Of the 293 patients studied, 227 (77.4%) had hydatid cysts only in the liver, while 55 (9.4%) had both liver and lung cysts. More than half of the liver cysts (65.9%) were located in the right portion of the liver (segment 5 to 8). Of the 293 cases, 52 (17.7%) underwent radical surgery, while 241 (82.3%) underwent conservative surgery. Recurrence of hydatid cyst was recorded in 46 (15%) of cases. Patients who were treated with radical surgery in comparison with those who had conservative surgery had a lower recurrence rate but a longer duration of hospital stay (P < 0.05). Conclusion: Recurrence remains as one of the major challenges in the management of hydatid cyst. Radical surgery reduces the chance of recurrence, although this procedure increases the length of hospital stay.
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Toxocariasis as a common neglected disease is the culprit of infecting all age groups. The current cross-sectional study was designed to assess the seroprevalence of toxocariasis and risk factors associated with seropositivity of Toxocara infection among the general population of adults in the Kavar district, south of Iran. A total of 1060 participants with an age range of 35 to 70 years from the Kavar region entered the study. Manual ELISA was utilized to detect anti-Toxocara-specific antibodies in their serum samples. In addition, demographic information as well as risk factors related to toxocariasis was collected from individuals who participated in the survey. The mean age of the participants was 48.9 (±7.9) years old. Out of 1060 subjects, 532 (50.2%) were men, and 528 (49.8%) were women. The overall Toxocara seroprevalence was 5.8% (61/1060). The prevalence of Toxocara seropositive cases was significantly different between males and females (p = 0.023). The seropositive rate for Toxocara infection was also significantly higher in housewives (p = 0.003) and subjects with learning disabilities (p = 0.008). Multivariable logistic regression revealed that housewives (OR = 2.04, 95% CI: 1.18-3.51, p = 0.010) and subjects with learning disability (OR = 3.32, 95% CI: 1.29-8.52, p = 0.013) were at increased risk of Toxocara infection. The findings of the current study depicted a noticeable seroprevalence of Toxocara infection in the general population in the Kavar district, southern Iran. An increased risk of toxocariasis has been associated with learning disabilities and being a housewife. All of the toxocariasis-positive cases had contact with animals, at some point in their life. In perspective, it is necessary to raise awareness of this infection among the population while surveilling Toxocara infection in high-risk groups.
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Cutaneous leishmaniasis (CL), a neglected tropical disease, is an important health problem in Fars Province, southern Iran. Fars, the fourth most populous Province in Iran, is the center of both anthroponotic and zoonotic cutaneous leishmaniasis (ZCL). Rodents, the reservoir of Leishmania major, play an important role in transmitting ZCL. In the present study, we report Leishmania infection in calomyscid rodents for the first time in mountainous residential areas of Shiraz, the capital of Fars Province, in southern Iran. Rodents were trapped in urban mountainous areas. The skin, liver, and spleen of rodents were examined microscopically for Leishmania infection. In addition, DNA was extracted from the tissues and they were evaluated for Leishmania infection by targeting the kDNA and subsequent sequencing of the nuclear rDNA internal transcribed spacer two (ITS2) region. DNA of L. major was detected in the spleen and liver of calomyscid rodents. Molecular evolution based on DNA-sequencing of the ITS2 gene confirmed the taxonomic situation of the parasite as L. major. Our findings suggest the eco-epidemiological importance of calomyscid rodents in the foci of leishmaniasis in the mountainous residential area on the plateau of Iran. These rodents may play a role in the transmission of leishmaniasis in a residential area and could be considered a potential reservoir for CL.
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Cystic echinococcosis (CE) is a common zoonotic infection in all provinces of Iran, especially in areas where people live on agriculture and animal husbandry. The current cross-sectional population-based study is aimed at determining the seroprevalence of CE in Kavar County, Fars province, southern Iran. Blood samples were collected from 1500 individuals (749 males and 751 females). Anti-hydatid cyst antibodies were detected, using a recombinant B8/1 antigen of Echinococcus graunlosus in an ELISA system. Multivariate logistic regression analysis was used to find out the independent risk factor for CE seropositivity. Anti-hydatid cyst antibodies were detected in the sera of 131 out of 1500 participants, corresponding to a seroprevalence rate of 8.73%. With a statistically significant difference (p < 0.05), the seroprevalence of hydatid cyst in males and females was 5% and 7%, respectively. Differences in the seropositivity of hydatid cysts were also statistically significant by occupation (p < 0.05). The seroprevalence of hydatid cyst was significantly higher in the age group of 35-45 years than in other age groups (p < 0.05). Multivariate logistic regression showed that only age was associated with seropositivity to CE (OR = 2.23, 95% CI: 1.33-3.72). Moreover, there was a statistically significant association between seropositivity to hydatid cysts and underlying diseases such as hypertension (p = 0.049) and fatty liver (p = 0.048). The findings of this study indicate that the seroprevalence rate of CE is relatively high in the Fars province, southern Iran, and this high rate of infection is mainly linked to people's jobs and lifestyles.
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Cistos , Equinococose , Echinococcus , Animais , Estudos Transversais , Equinococose/epidemiologia , Feminino , Humanos , Irã (Geográfico)/epidemiologia , Masculino , Fatores de Risco , Estudos SoroepidemiológicosRESUMO
Intellectually disabled individuals are more prone to parasitic infections due to their unusual behaviors, immune and nutrient deficiencies, and living conditions. The current study is aimed at evaluating the prevalence of intestinal parasites in institutionalized intellectually disabled individuals in Bandar Abbas County in the south of Iran. Subjects of the study were 119 individuals, living in an intellectually disabled individual care center. Demographic features of the subjects including age, sex, intellectual disability type, and duration of their stay in the center were recorded. A stool sample was taken from each subject and evaluated by direct wet mount and formalin-ethyl-acetate concentration methods. Samples were also examined, using trichrome and modified acid-fast permanent staining. The mean age of the subjects was 27.6 (±2.24), ranging from 4 to 60 years old. Of the 119 participants, 55 (46.2%) were male, and 64 cases (53.8%) were female. Overall, 31 individuals (26.1%, 95% CI: 18.4-34.9) were found to be infected with at least one type of intestinal parasite. Blastocystis hominis, as the most common detected parasite, was detected in 13 (10.1%), Entamoeba coli in 12 (10.1%), Giardia lamblia in 5 (4.2%), Cryptosporidium in 2 (1.7%), Iodamoeba butchlii in 2 (1.7%), and Endolimax nana in 1 (0.8%) of participants. Three cases had coinfection with Blastocystis hominis and Entamoeba coli, one case was infected with Blastocystis hominis and Giardia lamblia, and one case was coinfected with Giardia lamblia and Entamoeba coli. There were no statistically significant associations between intestinal parasitic infection and gender, age, type of intellectual disability, or duration of stay in the care center (P > 0.05). The findings of the present study indicate a relatively high prevalence of parasitic infections in people with intellectual disabilities in Bandar Abbas, southern Iran. Noteworthy is the high prevalence of Blastocyst and also the presence of Cryptosporidium infection in these people. Periodic treatment of these people and improvement of their maintenance conditions can be considered for the prevention and control of intestinal parasitic infection in these people.
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Background: Serological diagnosis of Strongyloides stercoralis (S. stercoralis) is fre-quently challenging because of cross-reactivity with other parasitic nematodes. Therefore, it is necessary to introduce novel serological tests with high performance to properly diagnose this neglected parasitic infection. The purpose of the current study was to design a multi-epitope construct for the diagnosis of S. stercoralis. Methods: For the purpose of this study, first, highly antigenic segments and potential immunodominant epitopes of S. stercoralis were identified from two antigenic proteins, and then all of the selected parts were linked by an appropriate linker. Next, the physicochemical features of the designed construct were analyzed. Then, tertiary structures of the construct were built and evaluated to find out the best one. Lastly, the amino acid sequence was reverse-translated and optimized for over-expression in Escherchia coli (E. coli). Results: The bioinformatic evaluation indicated that the designed protein construct could be hydrophilic, thermostable, and acidic and the estimated half-life was more than 10 hr in E. coli. Conclusion: According to the results of the study, the designed construct could be used as an efficient antigen in the ELISA system for serological diagnosis of human strong-yloidiasis.
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Giardia duodenalis is a well-known flagellated parasite and the causative agent of protozoal diarrhea in animals and humans worldwide. Current study was aimed at determination of G. duodenalis prevalence, genetic variation and zoonotic significance in various species of rodents in Shiraz, southwestern Iran. In brief, 120 fecal specimens were collected from rodents (Rattus rattus, Rattus norvegicus, and Mus musculus) during May up to November 2021 and microscopically examined for Giardia cysts. Further molecular characterization of positive samples was done by nested-PCR, followed by nucleotide sequencing of the triose phosphate isomerase (tpi) gene. A total prevalence of 3.3% (4/120) was observed in rodents, with highest rate in black rats [5% (2/40)]. Regarding brown rats and house mice, only one sample was found to be positive, showing 2.5% and 2.5% prevalence, respectively. It is noteworthy that Giardia B and G assemblages were found in black rats (one case/genotype), whereas the only positive samples from brown rats and house mice were characterized as assemblage G. The major findings of the present study were the presence of both zoonotic and non-zoonotic Giardia assemblages in examined rats in Shiraz and the potential of black rats to harbor Giardia infection to humans. These concerns should be taken seriously in terms of public health. Nevertheless, the true epidemiology and assemblage distribution of Giardia is still open to question.
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Giardia lamblia , Giardíase , Doenças dos Roedores , Animais , Fezes/parasitologia , Genótipo , Giardia lamblia/genética , Giardíase/epidemiologia , Giardíase/parasitologia , Giardíase/veterinária , Camundongos , Filogenia , Prevalência , Ratos , Doenças dos Roedores/epidemiologia , Roedores , Triose-Fosfato Isomerase/genéticaRESUMO
Sarcocystis species are intracellular protozoan which mostly complete their life cycle in two hosts. The parasite has a significant economic, medical and veterinary impact in many regions of the world and considered as a significant health problem in Iran. However, most of infections are asymptomatic and mortality is extremely rare. The present study aimed to determine the molecular phylogeny of the Sarcocystis species isolated from sheep slaughtered in southwest Iran, using mitochondrial DNA sequences of 18 S rRNA gene. The DNA was extracted from sheep muscular tissue (n = 60), and partial sequence of 18 S rDNA was amplified and sequenced. Phylogenetic analysis revealed two monophyletic clades representing S. moulei (n = 3) and Sarcocystis spp. (n = 3). BI posterior probability and MP bootstrap values strongly supported the monophyly of these clades. In conclusion, phylogenetic analysis of Sarcocystis species using 18 S rRNA gene could be helpful in identifying the new species of the Sarcocystis.
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Coronavirus 2019 (COVID-19) is a global concern for public health. Thus, early and accurate diagnosis is a critical step in management of this infectious disease. Currently, RT-PCR is routine diagnosis test for COVID-19, but it has some limitations and false negative results. enzyme-linked immunosorbent assay (ELISA) against SARS-CoV-2 antigens seems to be an appropriate approach for serodiagnosis of COVID-19. In the current study, an ELISA system, using a recombinant nucleocapsid (N) protein, was developed for the detection of IgM and IgG antibodies to SARS-CoV-2. The related protein was expressed, purified, and used in an ELISA system. Sera samples (67) for COVID-19 patients, as well as sera samples from healthy volunteers (112), along with sera samples from non-COVID-19 patients were examined by the ELISA system. The expression and purity of the recombinant N protein were approved by SDS-PAGE and Western blotting. The sensitivity of ELISA system was 91.04 and 92.53% for the detection of IgG and IgM antibodies, respectively. Moreover, the specificity of the developed ELISA system for IgG and IgM were 98.21 and 97.32%, respectively. Our developed ELISA system showed satisfactory sensitivity and specificity for the detection of antiSARS-CoV-2 IgM and IgG antibodies and could be used as a complementary approach for proper diagnosis of COVID-19.
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COVID-19 , SARS-CoV-2 , Humanos , Proteínas do Nucleocapsídeo , Imunoglobulina G , COVID-19/diagnóstico , Nucleocapsídeo , Ensaio de Imunoadsorção Enzimática , Sensibilidade e Especificidade , Proteínas Recombinantes , Imunoglobulina MRESUMO
Background. Toxoplasma gondii is an intracellular protozoan parasite responsible for systemic disease in a wide range of warm-blooded animals. The current study is aimed at evaluating the prevalence of Toxoplasma infection in dogs, using serological and molecular methods in rural areas in Kazeroun Township, Fars province, southern Iran. Methods. Blood samples were obtained from 60 clinically healthy dogs with an age range of 1 to 7 years in three rural areas of Fars province, southern Iran. Sera and buffy coats were used to assess the T. gondii infection using both modified agglutination test (MAT) and real-time PCR. Results. Antibodies against T. gondii were detected in 5 out of 60 (8.3%) dogs by the MAT method, and T. gondii DNA was detected in 17 out of 60 (28.3%) studied animals. There was no significant association between sex and seropositivity to Toxoplasma (p > 0.05). Fair agreement (kappa = 0.27) was seen between molecular and serological findings where three dogs with positive serological results had a positive molecular test. Conclusion. Findings of the present study show a relatively high prevalence of T. gondii infection in dogs in rural areas in Fars province, southern Iran. Finding the parasite genotype in dogs deserves further study.
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BACKGROUND: Different genotypes of Echinococcus granulosus sensu stricto (s.s.) isolated from livestock and humans have been identified based on cox1 and nad1 genomic fragments. The present study was performed to differentiate the G1/G3 genotypes of Echinococcus granulosus (s.s.) isolated from humans and livestock (sheep and cattle) from Azerbaijan in northwestern Iran, Fars Province in southern Iran, and Van province in Eastern Turkey, using the nad5 gene fragment as a suitable marker to distinguish these two genotypes. METHODS: A total of 60 pathologically confirmed human hydatid cysts and 90 hydatid cyst samples from livestock were collected from Turkey and Iran. PCR was performed on all of the samples, targeting the nad5 gene. Based on PCR product quality, host type, and the geographical area where the samples were obtained, 36 of the samples were sequenced and were used in the phylogenetic analysis. RESULTS: Out of 36 evaluated samples, 26 (72.2%) samples belonged to G1, and 10 (27.8%) samples belonged to the G3 genotype. Out of 21 samples from Turkey, 16 (76.2%) were G1 and 5 (23.8%) were G3, while out of 15 samples from Iran, 10 (66.7%) were G1 and 5 (33.3%) were the G3 genotype. None of the samples isolated from humans in Iran or from sheep in Turkey were G3. Overall, between the two countries, 18.18% of E. granulosus isolates in cattle, 41.66% of isolates in sheep, and 23.07% of human samples were identified as G3, and the others as the G1 genotype. The G3 genotype was not detected in human samples from Iran or sheep samples from Turkey. CONCLUSION: The findings of the study revealed that the G1 genotype of E. granulosus s.s. is the predominant genotype in humans and livestock, both in Turkey and Iran. The ratio of the E. granulosus s.s. G1 to G3 genotype was 3.2 in Turkey and 2 in Iran. The study also further confirmed that the nad5 gene properly differentiated the G1/G3 isolates of E. granulosus from both humans and livestock.
Assuntos
Doenças dos Bovinos/parasitologia , Equinococose/parasitologia , Echinococcus granulosus/genética , NADH Desidrogenase/genética , Doenças dos Ovinos/parasitologia , Animais , Bovinos , Doenças dos Bovinos/epidemiologia , Primers do DNA/genética , Equinococose/epidemiologia , Echinococcus granulosus/isolamento & purificação , Genes Mitocondriais/genética , Genótipo , Haplótipos , Humanos , Irã (Geográfico)/epidemiologia , Gado , Filogenia , Sensibilidade e Especificidade , Ovinos , Doenças dos Ovinos/epidemiologia , Turquia/epidemiologiaRESUMO
BACKGROUND: Cystic echinococcosis (CE) is one of the most important helminthic parasitic diseases in Iran. The current study aimed to assess the seroprevalence of CE in North Khorasan Province, Northeast of Iran in 2018. METHODS: The study was carried out in seven cities of North Khorasan Province. Venous blood samples were collected from 932 individuals referring to health centers of those seven cities. A questionnaire was used to obtain the data regarding the subject's gender, age, residence and risk factors linked to the hydatid cyst. Sera samples were evaluated for anti-hydatid cyst antibodies in an ELISA system, using a recombinant B8/1 antigen of E. granulosus. RESULTS: Of the 932 recruited subjects, 496 (53.2%) were male and 436 (46.8%) were female. The range of participants' age was between 11 to 83 yr old and the mean age of the subjects was 35.4 (±12.7) years. Anti-hydatid cyst antibodies were detected in the sera of 37 out of 932 subjects, corresponding to a seroprevalence rate of 3.96%. From these, 20 (54.05%) were male and 17 (45.95%) were female. There were no associations between seropositivity to hydatid cyst and age, the gender of the participants, residential areas and having contact with dogs (P>0.05). CONCLUSION: CE is relatively prevalent throughout the North Khorasan Province in the Northeast of Iran. Rate of CE infection in this Province is somewhat similar to the rate of infection in other parts of the country.