The Role of Dicrocoelium dendriticum Egg Antigen in Colitis: A Molecular, Pathological and Serological Study in an Experimental Model of C57BL/6 Mice.

BACKGROUND: Inflammatory bowel disease (IBD) is a chronic and recurrent disease of the gastrointestinal tract that enhances the chance of developing colorectal cancer. Since standard treatments such as Mesalazine have limited effectiveness and are often accompanied by numerous side effects, the use of immune modulators derived from worms has been proposed as a new immunotherapy method for inflammatory diseases such as ulcerative colitis. The aim of this study is to investigate the protective effects of D. dendriticum egg antigen on DSS-induced colitis in C57BL/6 mice. METHODS: D. dendriticum egg antigen was extracted and DSS (3.5%) was used to induce colitis in mice. Treatment and prophylaxis included intraperitoneal injections of D. dendriticum egg antigen. Histopathological indicators and the disease activity index (DAI), including weight loss, rectal bleeding, stool consistency, and rectal prolapse, were used to assess the severity of colitis. Real-time PCR measured the expression of transforming growth factor-ß (TGF-ß) and interleukin-17 (IL-17), while ELISA determined the concentration of these cytokines. RESULTS: Treatment with D. dendriticum egg antigen significantly improved the clinical symptoms and decreased the severity of DSS-induced colitis. Furthermore, D. dendriticum egg antigen increased the expression of TGF-ß mRNA and reduced the expression of IL-17 mRNA, leading to a positive adjustment in the regulation of proteins and reduction of inflammatory proteins. As a result, the macroscopic, microscopic inflammation and activity index (DAI) of DSS-induced decreased. CONCLUSION: D. dendriticum egg antigen provides a promising new way to modulate the immune system and improve ulcerative colitis.

Immunomodulatory effect of Dicrocoelium dendriticum ova on DSS-induced experimental colitis in C57BL/6 mouse.

Inflammatory bowel disease (IBD) significantly diminishes an individual's quality of life and increases the risk of colorectal cancer. Recent clinical and experimental findings suggest that infection with parasitic helminths may suppress the development of certain inflammatory conditions. The objective of this study was to evaluate the immunoregulatory effects of Dicrocoelium eggs on experimentally induced colitis in C57BL/6 mice using dextran sulfate sodium (DSS). C57BL/6 mice received 3.5% DSS orally for 7 days to induce colitis, during which they were treated intraperitoneally with Dicrocoelium eggs. The severity of colitis was assessed through parameters such as body weight, stool consistency or bleeding, disease activity index (DAI), colon lengths, macroscopic scores, histopathological findings, colon gene expression levels, and serum cytokine levels. Our results indicated that Dicrocoelium eggs administration significantly reduced the severity of colitis and disease activity. Histopathological scores improved, correlating with downregulation of IFN-γ and upregulation of IL-4 expression. This findings suggest the therapeutic potential of Dicrocoelium eggs in treating colitis. Immunotherapy involving Dicrocoelium eggs primarily induces a Th2 response and modulates IFN-γ, contributing to reduced inflammation in colitis. Thus, this approach could be a promising therapeutic strategy for alleviating inflammation in IBD.

Morphometric Characteristics and Species Identification of Bed Bugs (Hemiptera: Cimicidae) in Eastern Iran.

Background: The common bed bugs, Cimex lectularius, and tropical bed bugs, Cimex hemipterus are the primary species of public health importance in the family Cimicidae. This study aimed to determine the morphometric criteria and prevalent species of bed bugs in eastern Iran. Methods: Bed bugs were collected from March 2021 to June 2022 from infested residential buildings and apartments in eastern Iran, including North Khorasan, Khorasan Razavi, and Sistan and Baluchistan Provinces. The morphological and morphometrical characteristics were used to identify collected bed bugs at inter- and intra-specific levels. Results: A total of 34 isolates comprising 127 adult bed bugs were collected from Bojnord, Mashhad, Neishabur, Taibad, Sabzevar, Kashmer, Zahedan, Saravan, Rask, Pishin and Chabahar. Of these, 33 isolates (n=124) were found to be tropical bed bugs, C. hemipterus, and one population (n=3) was identified as C. lectularius. The index pronotal width/length ratio was calculated from 2.72 to 2.94 and 1.98 to 2.47 for C. lectularius and C. hemipterus, respectively. The length/width ratio of the hind femur was 3.365 in C. hemipterus and 4.267 in C. lectularius. The ratio of length/width of the third femur (F3 l/w) between populations of C. hemipterus was different, and this difference was statistically significant (P< 0.05). Conclusion: The results of this study indicated that C. hemipterus was the dominant bed bug species in the east of Iran and provided more morphometric criteria of C. hemipterus for researchers to identify the species and determine the intraspecific variations in the present and future.

Testing the Sensitivity of the Tropical Bed Bug Cimex hemipterus (Hemiptera: Cimicidae) to Deltamethrin, Phoxim and Propetamphos in Eastern Iran.

Background: Bedbugs are nocturnal ectoparasites that have made a comeback after 20 years and have become one of the main challenges for pest control methods worldwide. Monitoring chemical pesticide resistance is crucial for identifying the best bed bugs management strategies to effectively manage arthropods. This study aims to assess the susceptibility of Cimex hemipterus (Hemiptera: Cimicidae) collected from different parts of Khorasan-Razavi Province, (northeast of Iran) to deltamethrin (pyrethroid), phoxim (organothiophosphate) and propetamphos (phosphoramidate). Methods: This study was conducted from Dec 2020 to May 2021. The efficacy of three insecticides (deltamethrin, phoxim, and propetamphos) on adult C.hemipterus was assessed using a bioassay method recommended by the World Health Organization (WHO),. Concentrations of deltamethrin used were 10, 20, 40, 80, 160, 320, 640, 1280, and 2560 ppm, while concentrations of phoxim and propetamphos were 10, 40, 160, and 320 ppm. The bed bugs were continuously exposed to the insecticide for 24 hours, and mortality was assessed at regular intervals during the observation period. The concentration-response data were subjected to POLO-PC software and data were analyzed by the one-way and two-way ANOVA procedures. Results: The lethal concentration fifty values of deltamethrin, phoxim and propetamphos on the examined bed bugs were 0.551, 0.148 and 0.237 ppm, respectively. Insecticide effects of phoxim were significantly higher (P≤ 0.05) compared to each of either deltamethrin and propetamphos agents. Conclusion: The insecticide effects of phoxim against bed bugs were significantly higher compared to each of either deltamethrin or propetamphosinsecticides.

Toxoplasmosis Frequency Rate in Rheumatoid Arthritis Patients in Northeastern Iran.

Background: Toxoplasmosis is a zoonotic disease caused by the parasite Toxo-plasma gondii, a cosmopolitan intracellular parasite. It can be a risk factor for auto-immune diseases, including rheumatoid arthritis (RA). This study was designed to investigate the possible association between serological history of T. gondii infection and defined clinical manifestation of RA in Northeast of Iran. Methods: Overall, serum samples were collected from 50 RA patients and 40 healthy controls, from Qaem Hospital in Mashhad City, northeastern Iran in 2018. Seroprevalence of T. gondii infection was determined by ELISA. Results: The prevalence of anti -T. gondii IgG in RA patients 48% (24.50) was significantly higher than the control group 10% (4.40) (P <0.001). Erythrocyte sedimentation rate (ESR), anti-cyclic citrullinated peptide (anti-CCP) and (rheumatoid factor) RF levels between the RA and control groups (P <0.01). Control group were matched with patients for age, gender and living area. Conclusion: Given that a high correlation has been demonstrated between positivity rate of anti-T. gondii IgG and RA in Northeastern Iran, further studies will be necessary to clarify the pathogenesis of T. gondii among these patients.

Evaluation of the immunoregulatory effect of Dicrocoelium dendriticum eggs on inflammatory and anti-inflammatory cytokines in EAE model.

Experimental autoimmune encephalomyelitis (EAE) is an appropriate model for the study of the immunologic and pathologic mechanisms in multiple sclerosis (MS). According to the hygiene hypothesis, helminths can improve immunoregulation and have therapeutic effects on immune-mediated diseases. In this study, we used Dicrocoelium dendriticum (Dicrocoeliidae, Platyhelminthes) eggs for the evaluation of their prophylactic and treatment effects on EAE disease. D. dendriticum eggs were extracted. Female C57BL/6 mice were immunized with the specific antigen MOG35-55 , and then the egg extracts were utilized for prophylaxis and/or treatment. Clinical symptoms and other relevant parameters were assessed daily. The mRNA expression of transforming growth factor-ß (TGF-ß), interleukin-10 (IL-10), IL-6, IL-23 and IL-17 were assessed with a real-time polymerase chain reaction technique. Furthermore, secretion of TGF-ß and IL-17 cytokines were determined by enzyme-linked immunosorbent assay. Data indicated that clinical symptoms in prophylaxis and treatment groups were decreased significantly in comparison with the untreated control group (p < .001). Our results showed a significant decrease in IL-17, as well as an increase in TGF-ß cytokine in the treatment group compared to the EAE control group (p < .01). Furthermore, in the prophylaxis and treatment groups, the mRNA expression of disease-associated cytokines decreased and the mRNA expression of the anti-inflammatory cytokines increased. In this study, the D. dendriticum egg ameliorates the clinical symptoms of the EAE model through the modulation of related cytokines of Th17 and Treg cells. Therefore, using this parasite egg could be a new treatment for MS.

Prevalence and Seasonality of Adult and Arrested Larvae of Gastrointestinal Nematodes of Sheep from Mashhad City, Northeastern Iran.

Background: This study was conducted to determine the prevalence and intensity of nematode infections in sheep located in northeastern Iran from Apr 2018 to Mar 2019. Methods: Gastrointestinal nematodes of 300 sheep were inspected. The season of slaughter, anatomic location where the parasite was located, the animal's sex, infection prevalence and intensity were recorded. Seasonal differences in arrested larvae numbers also were assessed using Cochran's Q test. Results: Overall, 4,331 adult nematode specimens were collected. Among the examined sheep, 53% (159/300) were infected with one or more nematode species. Among infected sheep, 42.8% were infected with a single species of nematode, 26.4% were infected with two species of nematodes, and 30.8% were infected with three or more species of nematodes. Marshallagia marshalli (13.3%) was the most common nematode recovered from the abomasums of infected sheep, while Trichostrongylus vitrinus (4.6%) was commonly recovered from the small intestines, and Trichuris ovis (25.6%) was commonly recovered from the large intestines. In total, 463 arrested larvae were found in the abomasums of 7.5% of infected sheep and 104 arrested larvae were found in the small intestines of 8.8% of infected sheep. A significantly higher numbers of arrested larvae were found in summer compared to autumn (P<0.001). Conclusion: Intestinal parasites continue to be a problem for sheep in northeastern Iran and additional control measures need to be explored.

Impact of a health educational interventional program on reducing the head lice infestation among pupils in an elementary school of a sub-tropical region: a quasi-experimental study.

BACKGROUND: Pediculosis is an important social challenge that can be caused by human head louse, Pediculus humanus capitis. This infestation is cosmopolitan, especially in countries with low hygiene and sanitation. Regular health education classes for students and their parents are required to reduce this infestation in schools and families. This study aimed to evaluate the impact of a health education intervention program on reducing head lice among pupils of an elementary school. METHODS: In a quasi-experimental study, a total of 880 elementary school girls were screened for pediculosis in (2017-2018). The prevalence of pediculosis follow-up continued monthly until the end of the school year after a two months educational intervention course. Visual inspection was applied for initial diagnosis of infection. The suspected cases were confirmed by wood lamp and potassium hydroxide (KOH) microscopic examination. Also eight questions about diagnostic, epidemiology, treatment and prevention were answered in a questionnaire by 50 volunteer parents before and after the training intervention course, to evaluate their general knowledge and measure the effectiveness of learning. All data were statistically analyzed using SPSS software (version 20.0, SPSS, Inc., Chicago, IL, USA). McNemar's test was also applied to investigate the prevalence rate before and after the intervention. A p -value of less than 0.05 was considered as statistically significant. RESULTS: At the beginning of study, pre-intervention prevalence of pediculosis among pupils was 8.4% (49/594). The mean age in all pupils was 9.86 ± 1.83 years old and the most infestation was shown in fourth-grade students with 10 years old. Analysis of statistics demonstrated a significant difference between having infestation and the number of members in the families. Post-intervention phase led to a decreased prevalence of pediculosis in pupils to 3% (8/594) (p-value < 0.05). Based on statistical analysis of questions before and after the training in the questionnaire, a significant increase of parental knowledge was observed on the prevention program of pediculosis (p-value < 0.001). CONCLUSION: The prevalence of pediculosis was significantly reduced following the educational interventions in the school. The applied interventions may be implemented in other residual centers to get rid of this important infestation.

Validation of a mixture of rK26 and rK39 antigens from Iranian strain of Leishmania infantum to detect anti-Leishmania antibodies in human and reservoir hosts.

Mediterranean type of visceral leishmaniasis (VL) is a zoonotic parasitic infection. Some provinces of Iran are endemic for VL while other parts are considered as sporadic areas. This study aimed to assess a combination of recombinant K26 and rK39 antigens as well as crude antigen (CA), derived from an Iranian strain of L. infantum, compared to direct agglutination test (DAT) for the detection of VL in humans and domestic dogs as animal reservoir hosts of the disease. A combination of rK26 and rK39 antigens and also CA was evaluated using indirect ELISA on serum samples of 171 VL confirmed humans (n = 84) and domestic dogs (n = 87) as well as 176 healthy humans (n = 86) and domestic dogs (n = 90). Moreover, 36 serum samples of humans (n = 20) and canines (n = 16) with other potentially infectious diseases were collected and tested for finding cross- reactivity. The results of ELISA were compared to DAT, currently considered as gold standard for the serodiagnosis of VL. The sensitivity and specificity, positive predictive and negative predictive values were calculated compared to DAT. The positive sera had previously shown a positive DAT titer ≥ 1:800 for humans and ≥ 1:80 for dogs. Analysis was done by MedCalc and SPSS softwares. Using the combination of rK26 and rK39 in ELISA, a sensitivity of 95.2% and a specificity of 93.0% % were found in human sera at a 1:800 (cut-off) titer when DAT-confirmed cases were compared with healthy controls; a sensitivity of 98.9% and specificity of 96.7%% were found at a 1:80 (cut-off) titer compared with DAT. A good degree of agreement was found between the combined rK39 and rK26-ELISA with DAT in human (0.882) and dog serum samples (0.955) by kappa analysis (p < 0.05). The ELISA using the CA test showed 75% sensitivity in human and 93.1% in dog serum samples as well as 53.5% specificity in human and 83.3% in dog,s sera, respectively. The combination of rK26 and rK39 recombinant antigen prepared from Iranian strain of Leishmania infantum showed high accuracy for the serodiagnosis of VL in human and domestic dogs. Further extended field trial with a larger sample size is recommended.

Geographical patterns and mechanisms of Cimex lectularius Linnaeus, 1758, and Cimex hemipterus Fabricius, 1803 (Hemiptera: Cimicidae) resistance to insecticides: a systematic review and meta-analysis.

This study aimed to review published scientific literature on bed bugs in countries where insecticide resistance has been reported worldwide from 2000 to 2021. Electronic databases, including Scopus, PubMed, and WOS, were searched. Out of 606 articles found in the initial search, we selected 57 articles, of which 40 articles had reported on Cimex lectularius (C. lectularius), and 22 papers had reported on Cimex hemipterus (C. hemipterus). Most studies on insecticide resistance were carried out on C. lectularius in North America (14, 35%) and C. hemipterus in Asia (16, 72.7%). The most common method used to detect bed bug resistance to insecticides was toxicological bioassay with an overall random pooled effect size of 0.38 (95% CI: 0.23-0.53) in C. lectularius and 0.46 (95% CI: 0.27-0.65) in C. hemipterus. Resistance to pyrethroids was reported against C. lectularius with an overall pooled effect size of 0.75 (95% CI: 0.56-0.94) and C. hemipterus with an overall pooled effect size of 0.81 (95% CI: 0.57-0.93) in 33.40 (82.5%) and 19.22 (86.3%) published articles, respectively. A very high resistance level to pyrethroids in both studied species was observed, and resistance ratios at the highest level were 76389.3 and 315.5 in C. lectularius and C. hemipterus, respectively. Resistance mechanisms against pyrethroids were reported from most locations except Iran and Thailand, but these mechanisms were not studied in other insecticide groups. These reports indicate that chemical control options for bed bugs are limited. Therefore, a combination of chemical and non-chemical strategies is recommended for bed bug control.

Spatio-temporal visualisation of cutaneous leishmaniasis in an endemic, urban area in Iran.

INTRODUCTION: Cutaneous Leishmaniasis (CL) is a significant public health concern worldwide. Iran is among the most CL-affected countries, being one of the six most endemic countries in the world. This study aimed to provide a spatio-temporal visualisation of CL cases in an endemic urban area in north-eastern Iran identifying high-risk and low-risk areas during the period 2016-2019. METHODS: This ecological study was conducted in the city of Mashhad, north-eastern Iran. All cases (n=2425) were diagnosed based on clinical findings and parasitological tests. The patient data were aggregated at the census tract level (the highest resolution available). CL incidence rates were subjected to Empirical Bayesian smoothing across the census tracts followed by spatial autocorrelation analysis to identify clusters and outliers. Spatial scan statistic was used to explore the purely temporal, purely spatial and spatio-temporal trend of the disease. In all instances, the null hypothesis of no clusters was rejected at p ≤0.05. RESULTS: The overall crude incidence rate decreased from 34.6 per 100,000 individuals in 2016 to 19.9 per 100,000 in 2019. Cluster analysis identified high-risk areas in south-western Mashhad and low-risk areas in the north-eastern areas. Purely time scan statistics identified March to July as the time period with highest risk for CL occurrence. One most likely purely high-risk spatial cluster and six secondary purely high-risk spatial clusters were identified. Further, two spatio-temporal high-risk clusters, one in the north of the city from April to August and a second in the south-western part from March to September were observed. CONCLUSIONS: Significant spatial, temporal and spatio-temporal patterns of CL distribution were observed in the study area, which should be considered when designing tailored interventions, such as effective resource allocation models, informed control plans and implementation of efficient surveillance systems. Furthermore, this study generated new hypotheses to test potential relationships between socio-economic and environmental risk factors and incidence of CL in high-risk areas.

Frequency of Toxocara Antibodies in Patients Clinically Suspected to Ocular Toxocariasis, Northeast of Iran.

BACKGROUND: Human toxocariasis is a neglected parasitic disease in most countries including Iran. Among different clinical forms of toxocariasis, ocular toxocariasis (OT) is an important disease resulting in severe vision loss. However, the prevalence and incidence of OT are currently unclear in Iran. This study aimed to determine the prevalence of ocular toxocariasis among patients with uveitis in the Northeast of Iran. METHODS: From 2015 to 2017, 510 patients with uveitis referred to Khatam-al-Anbia, a tertiary eye hospital at Mashhad, Iran were examined for OT. Serum samples of the suspected patients were obtained and evaluated for IgG against Toxocara canis using ELISA test. Anti-Toxocara IgG positive serums were further investigated using confirmatory Western blotting (WB) analysis. RESULTS: Twenty patients had pathologic changes and clinical presentations in the anterior and posterior segments of their eyes and they were clinically diagnosed ocular toxocariasis. Among the 20 patients, 2 (10%) patients showed IgG antibody against Toxocara canis on ELISA as well as on WB test. The calculated prevalence of ocular toxocariasis was about 0.4%. CONCLUSION: Ocular toxocariasis can be diagnosed both clinically and serologically in Mashhad, northeastern Iran. Although OT is a rare pathologic eye disease, it should be considered as one of the important cause of infectious posterior uveitis.

Local and systemic gene expression levels of IL-10, IL-17 and TGF-ß in active ocular toxoplasmosis in humans.

BACKGROUND: To compare mRNA expression of interleukin 10 (IL-10), interleukin 17 (IL-17) and Transforming Growth Factor-ß (TGF-ß) in aqueous humor (AH) and peripheral blood mononuclear cells (PBMCs) in human ocular toxoplasmosis (OT) and controls. METHOD: RNA isolation, cDNA synthesis and real-time polymerase chain reaction were performed on AH sediments and PBMCs of 16 patients with active OT and 21 controls at the Khatam-al-Anbia Eye Hospital, Iran. For comparison, Mann Whitney U test was used at a discrimination level of p < 0.05. Pearson and Spearman rank correlation test were applied for correlation with clinical parameters. RESULTS: The expression for IL-10 and IL-17 in the AH was 3.7- and 88.0-fold higher in OT than in controls (P = 0.04 and P = 0.03, respectively) whereas that of TGF-ß was 7.7-fold lower (P < 0.001). The expression levels for these cytokines in PBMC followed a similar pattern (IL-10 13.8-fold down-regulated (P = 0.001), IL-17 with 1.9-fold insignificantly upregulated (p = 0.43), TGF-ß 452.8-fold down-regulated (P = 0.002). Compared to PBMC, IL-10 coding mRNA was 1876-fold higher in the almost cell-free AH in OT (39.2-fold in controls), IL-17 coding mRNA was 9.4-fold higher (17.7-fold down-regulated in controls), and that coding for TGF-ß 207-fold higher in OT (7x105-fold in controls). The expression for IL-10, IL-17 and TGF-ß in AH thus followed an opposite pattern compared to that in PBMC. CONCLUSION: OT induces a highly-specific local immunoregulatory process as evidenced by an intraocular up-regulation of IL-10 and down-regulation of TGF-ß mRNA. This could indicate an attempt to prevent unnecessary tissue damage which is in line with a moderate local mRNA up-regulation for IL-17 which seems sufficient to control parasite proliferation. That this regulation is opposite to that in PBMC may be linked to intraocular immune deviation in the course of disease.

Spatio-temporal epidemiology of the tuberculosis incidence rate in Iran 2008 to 2018.

BACKGROUND: Effective reduction of tuberculosis (TB) requires information on the distribution of TB incidence rate across time and location. This study aims to identify the spatio-temporal pattern of TB incidence rate in Iran between 2008 and 2018. METHODS: This cross-sectional study was conducted on aggregated TB data (50,500 patients) at the provincial level provided by the Ministry of Health in Iran between 2008 and 2018. The Anselin Local Moran's I and Getis-Ord Gi* were performed to identify the spatial variations of the disease. Furthermore, spatial scan statistic was employed for purely temporal and spatio-temporal analyses. In all instances, the null hypothesis of no clusters was rejected at p ≤ 0.05. RESULTS: The overall incidence rate of TB decreased from 13.46 per 100,000 (95% CI: 13.19-13.73) in 2008 to 10.88 per 100,000 (95% CI: 10.65-11.11) in 2018. The highest incidence rate of TB was observed in southeast and northeast of Iran for the whole study period. Additionally, spatial cluster analysis discovered Khuzestan Province, in the West of the country, having significantly higher rates than neighbouring provinces in terms of both total TB and smear-positive pulmonary TB (SPPTB). Purely temporal analysis showed that high-rate and low-rate clusters were predominantly distributed in the time periods 2010-2014 and 2017-2018. Spatio-temporal results showed that the statistically significant clusters were mainly distributed from centre to the east during the study period. Some high-trend TB and SPPTB statistically significant clusters were found. CONCLUSION: The results provided an overview of the latest TB spatio-temporal status In Iran and identified decreasing trends of TB in the 2008-2018 period. Despite the decreasing incidence rate, there is still need for screening, and targeting of preventive interventions, especially in high-risk areas. Knowledge of the spatio-temporal pattern of TB can be useful for policy development as the information regarding the high-risk areas would contribute to the selection of areas needed to be targeted for the expansion of health facilities.

Evaluation of zoonotic platyhelminthe infections identified in slaughtered livestock in Iran, 2015-2019.

BACKGROUND: Platyhelminth infections of livestock can result in considerable economic losses. This study aims to evaluate the spatial frequency of cystic echinococcosis (CE), dicrocoeliasis, and fascioliasis in livestock slaughtered in Iran during the years 2015-2019 and estimate direct costs associated with organ condemnation due to these parasites. METHODS: Abattoir data from 413 abattoirs representing all 31 Iranian provinces were collected from the Iran Veterinary Organization. Infection prevalence was calculated per year at the province level. The Local Moran's I statistic was performed to evaluate spatial autocorrelation of animals positive at slaughter for the years 2015-2019. Direct costs associated with condemned livers were calculated for each parasitic condition, with costs associated with condemned lungs also included for CE. RESULTS: Overall prevalence values for the study timeframe were as follows: sheep and goat fascioliasis 1.56% (95% CI: 1.56-1.56%), cattle fascioliasis 3.86% (95% CI: 3.85-3.88%), sheep and goat dicrocoeliasis 4.63% (95% CI: 4.62-4.63%), cattle dicrocoeliasis 3.08% (95% CI: 3.07-3.09%), sheep and goat CE 5.32% (95% CI: 5.32-5.33%), and cattle CE 7.26% (95% CI: 7.24-7.28%). Northwest Iran had the highest prevalence of CE and fascioliasis. High infection areas for Dicrocoelium spp. included the provinces of Zanjan, Gilan, Qazvin, and Tehran, which are located in northern Iran. Direct economic losses for sheep and goat fascioliasis, dicrocoeliasis, and CE for the study period were US$13,842,759, US$41,771,377, and US$22,801,054, respectively. Direct economic losses for cattle fascioliasis, dicrocoeliasis, and CE for the study period were US$1,989,200, US$1,668,986, and US$2,656,568, respectively. CONCLUSION: Our findings provide valuable data for future monitoring of these important parasitic diseases in Iranian livestock. Disease control strategies are required to reduce the economic and public health impact of these platyhelminths.

Diagnosis of Acanthamoeba keratitis in Mashhad, Northeastern Iran: A Gene-Based PCR Assay.

BACKGROUND: The genus Acanthamoeba is a free-living opportunistic protozoan parasite, which widely distributed in soil and fresh water. Acanthamoeba keratitis, which causes a sight-threating infection of the cornea, is going to rise in Iran and worldwide. The aim of this study was to compare direct microscopy, culture and PCR for detection of Acanthamoeba spp. in clinical samples and to determine the genotypes of Acanthamoeba spp. by sequencing 18SrRNA gene. METHODS: Among patients clinically suspected to AK referred to a tertiary ophthalmology center at Mashhad, northeastern Iran. During 2017-18, twenty corneal scrapes specimens obtained. The samples were divided into three parts, subjected to direct microscopic examination, culture onto non-nutrient agar and PCR technique. Sensitivity, specificity, accuracy and likelihood ratio were evaluated. RESULTS: Among 20 persons clinically suspected to amoebic keratitis, 13(69.2%) patients definitely diagnosed as Acanthamoeba keratitis. Wearing contact lens, eye trauma due to foreign particle and swimming in fresh water were the main predisposing factors. Most of patients suffered from pain and photophobia. Corneal ring infiltration and epithelial defect were common clinical sings. Direct examination had the lowest sensitivity and sensitivity of both Nelson-PCR and JDP-PCR methods were equal and highest. In addition, the results of sequencing identified that all strains belonged to T4 genotype. CONCLUSION: Amoebic keratitis is a sporadic parasitic keratitis, which is mainly seen in contact lens user in Mashhad. PCR based on 18S ribosomal DNA with JDP primers is a reliable and highly sensitive method for diagnosis of Acanthamoeba keratitis in clinically suspected cases.

Anisakis allergy: unjustified social alarm versus healthy diet; commentary to the "Letter to the Editor" of Drs Daschner, Levsen, Cipriani, and del Hoyo, referencing to "World-wide prevalence of Anisakis larvae in fish and its relationship to human allergic anisakiasis: a systematic review".

Anisakiasis is an underrecognized condition globally, and accurate diagnosis remains problematic even in countries where the condition is well known. Our "systematic review" was conducted according to Prisma guidelines. The stated basis of our study was "syndromic surveillance." Both methods are recognized in published literature as valid to identify or predict disease and to make accessible large amounts of evidence from published literature. Our study identified Anisakis allergy "hot spots" and other geographical areas where fish are highly infected with Anisakis without commensurate studies of human allergy. Results of our study will open up new lines of enquiry. Norway, used as an example to discredit the scientific integrity of our article, has a cuisine thriving with raw fish dishes and many sushi restaurants. The peer reviewed data sets, confirmed A. simplex sensitization among the Norwegian population, although this has been overlooked by the authors of the "Letter to the Editor." The identification of hot spots in our study may be influential in many ways not the least in raising diagnostic suspicion to expedite accurate diagnosis.

World-wide prevalence of Anisakis larvae in fish and its relationship to human allergic anisakiasis: a systematic review.

The infective stage of Anisakidae nematodes responsible for allergic reactions in humans is found in a variety of edible fish and cephalopods. The identification of geographical regions that are high risk for infected seafood may help prevent allergic reactions in humans. Despite an abundance of published literature which has identified anisakid larvae in an array of edible seafood as well as scattered reports of human allergic anisakiasis, the relationship between the two has not been fully explored. Therefore, a systematic spatio-temporal study was conducted to determine the prevalence of Anisakis spp. in fish from January 2000 to August 2020 firstly to explore the relationship between fish infection and cases of allergic anisakiasis and secondly to use fish infection data to map potential allergic anisakiasis 'hot spots'. A systematic literature search for original English text articles was conducted through search engines, Web of Science, Scopus, PubMed, Science Direct and Google Scholar. Out of 3228 articles which describe anisakid infection in fish, 264 were used for data extraction. Of 904 articles describing allergic anisakiasis, 37 were used for data extraction. A qualitative summary of the extracted data was performed using equal interval method (ArcMap software) in order to compare the global distribution of Anisakis-infected fish. Of the 152-identified fish hosts, five families were most commonly infected with Anisakis spp. These included Lophiidae (86.9%), Trichiuridae (77.05%), Zeidae (70.9%), Merlucciidae (67.8%) and Gadidae (56.8%). The hot spot areas for allergic anisakiasis were North and northeast of Atlantic Ocean, southwest of USA, west of Mexico, south of Chile, east of Argentina, Norway, UK and west of Iceland (confidence 99%). The highest rate of allergic anisakiasis was in Portugal and Norway with the prevalence rate of 18.45-22.50%. Allergologists should consider allergic anisakiasis as a public health issue particularly in high-risk countries where high prevalences in fish have been demonstrated.

Effect of Low-level Laser Therapy With Different Locations of Irradiation on Postoperative Endodontic Pain in Patients With Symptomatic Irreversible Pulpitis: A Double-Blind Randomized Controlled Trial.

Introduction: This double-blind, placebo-controlled, clinical trial aimed to investigate the analgesic efficacy of low-level laser therapy (LLLT) with two different locations, and their comparison, in postoperative endodontic pain (PEP) levels in molars diagnosed with symptomatic irreversible pulpitis. Methods: Seventy-five patients with a molar tooth, diagnosed with symptomatic irreversible pulpitis, were divided into three groups of placebo, buccal only irradiation (BI), and buccal and lingual irradiation (BLI), with 25 cases being in each group. The participants received similar single-visit nonsurgical endodontic treatments. Then, a sham laser was used in the control group instead of LLLT. Individuals in BI and BLI groups received 80-second irradiation on the buccal surface and 80-second irradiation on each of the buccal and lingual surfaces respectively. A laser with an 808 nm wavelength, power of 100 mW, and a fiber diameter of 600 µm was used. PEP was assessed using a 0-100 mm VAS 4, 8, 24, and 48 hours after the treatment. Results: BLI showed a significantly higher reduction of PEP compared to placebo in all time intervals of this study. BLI was significantly more effective than BI 8 hours after the treatment. However, intragroup differences between BLI and BI groups at other time intervals and between BI and placebo groups in all time intervals were not significant. The number of taken analgesics in the BLI group was significantly lower than the placebo group and was on a statistical borderline compared to the BI group. Conclusion: LLLT with BLI was an effective measure as a supplement to oral analgesics in the reduction of PEP compared to the placebo.