The development of an integrated BIM-based visual demolition waste management planning system for sustainability-oriented decision-making.

In light of the suboptimal resource efficiency persisting in current demolition waste management (DWM) practices caused by inattentive and profit-driven decision-making due to the lack of tailored stringent legislation, monetary incentives, and benchmarking frameworks, this study aims to facilitate sustainability-oriented decision-making at the demolition planning stage. A practical Building Information Modelling (BIM)-based visual DWM planning system is designed, wherein the system seamlessly accommodates inventory analysis and Multi-Criteria Decision-Aiding (MCDA) algorithms into various interconnected modules. Moreover, this research proposes bespoke algorithms and colour coding schemes to quantify and visualise the recycling value of building components for augmenting the visual guidance of sustainable building design and selective demolition planning. Furthermore, a pilot case study demonstrates the system's applicability in a real-world demolition project. The findings unveil that improving the recycling rate substantially offsets carbon emissions and demolition waste disposal expenditures. The increment in beneficial impacts outweighs the additional energy consumption and costs for implementing sustainable DWM strategies based on the predefined geographical settings. This BIM-based system reforms the conventional demolition planning and DWM decision-making workflow by tackling technical barriers concerning data richness, interoperability, and result interpretation. It equips the users with intuitive visual design guidance and parallel scenario analysis when crafting sustainability-oriented DWM schemes. In summary, this research contributes to familiarising industry practitioners with sustainable DWM schematic design and circular economy principles. Moreover, it prompts the development of customised BIM libraries as repositories for updating and capitalising on DWM-related information that can be adapted to different regional contexts.

Molecular characterization of PMQR carrying bacteriophages in effluent discharge samples of Azerbaijan hospitals.

Quinolones are one of the most widely used drugs in medicine. Resistance to this agent has been increased significantly among the nosocomial isolates. The objective of this research was to study generalized transduction, as a potential mechanism for plasmid-mediated quinolone resistance (PMQR) genes acquisition among hospital effluent isolates. Discharge samples from hospital effluent were taken from four medical centers in Azerbaijan. Resident phages were enriched against resident enterobacterial hosts using standard phage enrichment protocols. Polymerase chain reaction (PCR) was used to examine phage stocks and bacterial isolates for the presence of PMQR determinants. All positive bacterial isolates for target genes were subjected to transduction assays. Restriction fragment length polymorphism (RFLP) profiles were determined for cluster analysis. A total of 55 pure phage stocks were prepared from 42 effluents. A total of 95 non-duplicated Gram-negative bacteria were isolated. Thirty-two EcoRV-RFLP profiles were determined for the 40 Escherichia coli phage stocks. Twenty-six of 40 (65%) E. coli phages were positive for qnrB (n = 15), qnrD (n = 7), qnrA (n = 3), and qnrC (n = 2) genes. A total of 34 (35.7%) bacterial isolates were recognized to have any PMQR genes including qnrB (n = 23), qnrD (n = 8), qnrA (n = 5), and qnrC (n = 3) genes. Present research provided a strong evidence for potential role of generalized transduction in persistence and circulation of PMQR genes in health care settings of Azerbaijan.

Identifying and prioritizing sustainability indicators for China's assessing demolition waste management using modified Delphi-analytic hierarchy process method.

Addressing the sustainability issues arising from construction and demolition waste management (DWM) has gained little traction due to the lack of incentives, stringent regulations, and systematic guidance. This study aims to empower systematic decision-making concerning DWM alternative selection by developing a sustainability assessment framework by coupling a modified Delphi method with the multicriteria decision analysis technique. First, the study identifies a comprehensive inventory of indicators across three dimensions of sustainability in the context of DWM. Next, the study combines a modified Delphi method with the analytic hierarchy process to validate and prioritize the selected sustainability indicators. For the first time, insights regarding the DWM sustainability indicators from China's construction industry practitioners' perspectives are elicited using a mixed method comprising online semistructured interviews and two rounds of questionnaire surveys. Experts participating in the research are mostly based in Guangzhou and Shenzhen, where local governments exhaust all efforts in promoting carbon-neutral and sustainable development. The findings reveal that eight sustainability indicators were regarded as the determinants for the sustainability performance of DWM, with the global warming potential (32%), energy efficiency (16.1%) and land use (13.5%) receiving the highest preference scores (weights) based on the experts' judgment. Notably, the economic factors like the total cost (6.54%) appeared not highly prioritized by the local experts as typically did in the previous studies from developing countries.

First molecular detection of SARS-CoV-2 virus in cockroaches.

Coronavirus is one of the main pathogens that primarily targets the human respiratory system. There are several ways to transmit this virus, such as direct contact or droplets spread by coughing or sneezing, and direct contact with fomites and surfaces is another way. This cross-sectional study was conducted in Shiraz, southern Iran, in 2021. 5 locations, including 3 hospitals and 2 dormitories, were selected for the survey. The cockroaches were collected from selected locations and transferred to the Laboratory of Medical Entomology at Shiraz University of Medical Sciences. All specimens were identified morphologically. The external and gastrointestinal washouts of collected samples with sterile phosphate-buffered saline separately were used for molecular analysis. An RT-qPCR assay, which suggests the possible insect­borne transmission, was used. External and gastrointestinal washout of B. germanica from Dastgheyb Dormitory and P. americana from Ali-Asghar Hospital were positive for contamination with the SARS-CoV-2. Cockroaches spread the virus in the environment and contaminate human food and various surfaces of buildings. Their role will be more important in crowded places such as hotels, lodging houses, restaurants, and hospitals; vector control programs should be carried out with more accuracy in such places.

Acceleration in healing of infected full-thickness wound with novel antibacterial γ-AlOOH-based nanocomposites.

This study was conducted to synthesize γ-AlOOH (bohemite)-based nanocomposites (NCs) of Au/γ-AlOOH-NC and its functionalized derivative using chitosan (Au/γ-AlOOH/Ctn-NC) and with the help of one-step Mentha piperita. The physicochemical characteristics of the NCs were investigated. In addition, biomedical properties, such as antibacterial activity under in vitro and in vivo conditions, and cell viability were assessed. Wound healing activity on infected wounds and histological parameters were assessed. The gene expressions of TNF-α, Capase 3, Bcl-2, Cyclin-D1 and FGF-2 were investigated. The TEM and FESEM images showed the sheet-like structure for bohemite in Au/γ-AlOOH-NC with Au nanoparticles in a range of 14-15 nm. The elemental analysis revealed the presence of carbon, oxygen, aluminum, and Au elements in the as-synthesized Au/γ-AlOOH. The results for toxicity showed that the produced nanocomposites did not show any cytotoxicity. Biomedical studies confirmed that Au/γ-AlOOH-NC and Au/γ-AlOOH/Ctn-NC have anti-bacterial properties and could expedite the wound healing process in infected wounds by an increase in collagen biosynthesis. The administration of ointment containing Au/γ-AlOOH-NC and Au/γ-AlOOH/Ctn-NC decreased the expressions of TNF-α, and increased the expressions of Capase 3, Bcl-2, Cyclin-D1 and FGF-2. The novelty of this study was that bohemite and Au nanoparticles can be used as a dressing to accelerate the wound healing process. In green synthesis of Au/γ-AlOOH-NC, phytochemical compounds of the plant extract are appropriate reagents for stabilization and the production of Au/γ-AlOOH-NC. Therefore, the new bohemite-based NCs can be considered as candidate for treatment of infected wounds after future clinical studies.

Methotrexate loaded in alginate beads for controlled drug release against breast cancer.

Methotrexate (MTX), as a folate antagonist is used for breast cancer chemotherapy, but its application due to the adverse side effects was limited. In this study, MTX were encapsulated in magnetic alginate beads coated with glutaraldehyde to control its release in order to reduce the side effects and improve its stability. The complex was characterized by physicochemical studies. The encapsulation efficiency was 75 % and the complex showed acceptable controlled release behavior. The cell cytotoxicity assessed using methylthiazol tetrazolium (MTT) method showed that magnetic alginate beads-MTX, in lower dosage has higher anticancer effect compared to the free MTX. The real-time polymerase chain reaction (PCR) was used to evaluate apoptotic factors Bcl2 associated X gene (Bax), B-cell lymphoma 2 (Bcl-2), and neuroinflammatory marker tumor necrosis factor-alpha (TNF-α) genes expression level on the treated cells. The findings demonstrated the significant increase of expression of Bax and a significant decrease in the expressions of Bcl-2 and TNF-α in Michigan cancer foundation-7 (MCF-7) cells. These results indicated that the developed drug can overcome the side effects of MTX and offer a controlled drug release for a sustained period with the long-term treatment of breast cancer.

Theileria equi in the horses of Iran: Molecular detection, genetic diversity, and hematological findings.

In all equids worldwide, Theileria equi and Babesia caballi are believed to be two important erythrocytic protozoa that cause equine piroplasmosis. In addition, it was recently discovered that Theileria haneyi is another potential equine piroplasmosis (EP) agent. Ixodid ticks are the major vectors of these parasites. Equine piroplasmosis is of international importance and affects enormously the equine industry. In this study, for the first time, molecular prevalence and genetic diversity of piroplasma parasites (T. equi and B. caballi) in horses from Fars province (south of Iran) were determined. Also, hematological alterations of naturally infected horses were analyzed. PCR positive horses showed anemia, thrombocytopenia, leukocytosis with a left shift of neutrophilia, and monocytosis. PCR results revealed that, from 133 blood samples of horses, 40 samples were positive (30.07%). The occurrence of T. equi in this area (30.07%) was more than the national average prevalence of T. equi (24.11%), but B. caballi prevalence in study area (0%) was less than the average of previous studies in Iran (5.47%). Our findings revealed that the T. equi was widespread in Fars province of Iran. PCR products of 18S rDNA and EMA-1 genes of T. equi strains were sequenced successfully. All 18S rDNA sequences collected in this experiment revealed 100% similarity together. According to the phylogenetic tree constructed using the 18S rDNA gene, Iranian T. equi is clustered with strains from Cuba (KY111762, KY111761) and USA (CP001669, JX177672). So, this could be concluded that T. equi studied in this research, and those strains are initiated from a common T. equi ancestor at an unknown time ago. Also, the phylogenetic tree based on EMA-1 gene demonstrated a genetically diverse population of Iranian T. equi strains (10 different genotypes). As EMA-1 is one of the most immunogenic antigens in this parasite, such variability could be a concern about the efficacy of T. equi vaccines. Finally, more studies on equine piroplasmosis in the provinces of the southern region of Iran are recommended to create a better vision of disease in this region.

Cutaneous leishmaniasis in Iran: A review of epidemiological aspects, with emphasis on molecular findings.

Leishmania parasites can cause zoonotic cutaneous leishmaniasis (CL) by circulating between humans, rodents, and sandflies in Iran. In this study, published data were collected from scientific sources such as Web of Science, Scopus, PubMed, Springer, ResearchGate, Wiley Online, Ovid, Ebsco, Cochrane Library, Google scholar, and SID. Keywords searched in the articles, theses, and abstracts from 1983 to 2021 were cutaneous leishmaniasis, epidemiology, reservoir, vector, climatic factors, identification, and Iran. This review revealed that CL was prevalent in the west of Iran, while the center and south of Iran were also involved in recent years. The lack of facilities in suburban regions was an aggravating factor in the human community. Some parts of southern Iran were prominent foci of CL due the presence of potential rodent hosts in these regions. Rhombomys opimus, Meriones lybicus, and Tatera indica were well-documented species for hosting the Leishmania species in Iran. Moreover, R. opimus has been found with a coinfection of Leishmania major and L. turanica from the northeast and center of Iran. Mashhad, Kerman, Yazd, and sometimes Shiraz and Tehran foci were distinct areas for L. tropica. Molecular identifications using genomic diagnosis of kDNA and ITS1 fragments of the parasite indicated that there is heterogeneity in leishmaniasis in different parts of the country. Although cutaneous leishmaniasis has been a predicament for the health system, it is relatively under control in Iran.

Title: Leishmaniose cutanée en Iran : une synthèse des aspects épidémiologiques, mettant l'accent sur les découvertes moléculaires. Abstract: Les parasites Leishmania peuvent établir une leishmaniose cutanée zoonotique (LC) en circulant entre les humains, les rongeurs et les phlébotomes en Iran. Dans cette étude, les données publiées ont été collectées à partir de ressources scientifiques telles que Web of Science, Scopus, PubMed, Springer, ResearchGate, Wiley Online, Ovid, Ebsco, Cochrane Library, Google Scholar et SID. Les mots-clés recherchés dans les articles, les thèses et les résumés de 1983 à 2021 étaient leishmaniose cutanée, épidémiologie, réservoir, vecteur, facteurs climatiques, identification et Iran. Cet examen a révélé que la LC est répandue dans l'ouest de l'Iran, tandis que le centre et le sud de l'Iran sont également impliqués ces dernières années. Le manque d'équipements dans les régions suburbaines est un facteur aggravant dans la communauté humaine. Certaines parties du sud de l'Iran sont des foyers importants de LC en raison de la présence d'hôtes potentiels de rongeurs dans ces régions. Rhombomys opimus, Meriones lybicus et Tatera indica sont des espèces bien documentées pour héberger les espèces de Leishmania en Iran. De plus, R. opimus a été trouvé avec une co-infection de Leishmania major et L. turanica au nord-est et au centre de l'Iran. Mashhad, Kerman, Yazd et parfois des foyers de Shiraz et de Téhéran sont des zones distinctes pour L. tropica. Les identifications moléculaires utilisant le diagnostic génomique des fragments d'ADNk et ITS1 du parasite ont indiqué qu'il existe une hétérogénéité dans la leishmaniose dans différentes parties du pays. Bien que la leishmaniose cutanée ait été une situation difficile pour le système de santé, elle est relativement contrôlée en Iran.

Molecular-based detection of Leishmania tropica isolates among sensitive, resistant, and relapsed patients treated with Meglumine Antimoniate.

The appearance of resistance to pentavalent antimony, as the mainline of treatment for Cutaneous Leishmaniasis (CL) has been reported from Iran. According to the patients' laboratory and clinical history, 96 archived slides of patients infected with Leishmania tropica (L. tropica) treated with Meglumine Antimoniate (Glucantime®) were selected. After microscopic examination, Nested Polymerase Chain Reaction (Nested-PCR) and Restriction Fragment Length Polymorphism (RFLP) assays were done for each sample. In Nested-PCR, all positive samples were characterized as L. tropica. Additionally, some positive products of sensitive, resistant, and recidivans cases were selected to check their differentiations by sequencing software. In RFLP, various patterns of schizodemes were detected according to the reference patterns. Most sensitive cases of L. tropica (treated with Glucantime®) were categorized as schizodeme B, and most resistant cases were identified as schizodeme B and D. In recidivans cases, 91% of specimens categorized as schizodeme A and B. However, study on the type of L. tropica isolates that are resistant or sensitive to Glucantime® could be helpful before drug therapy.

Region-income-based prioritisation of Sustainable Development Goals by Gradient Boosting Machine.

The Sustainable Development Goals (SDGs) seek to address complex global challenges and cover aspects of social development, environmental protection, and economic growth. However, the holistic and complicated nature of the goals has made their attainment difficult. Achieving all goals by 2030 given countries' limited budgets with the economic and social disruption that the COVID-19 pandemic has caused is over-optimistic. To have the most profound impact on the SDGs achievement, prioritising and improving co-beneficial goals is an effective solution. This study confirms that countries' geographic location and income level have a significant relationship with overall SDGs achievement. This article applies the Gradient Boosting Machine (GBM) algorithm to identify the top five SDGs that drive the overall SDG score. The results show that the influential SDGs vary for countries with a specific income level located in different regions. In Europe and Central Asia, SDG10 is among the most influential goals for high-income countries, SDG9 for upper-middle-income, SDG3 in low and lower-middle-income countries of Sub-Saharan Africa, and SDG5 in Latin America and the Caribbean upper-middle-income countries. This systematic and exploratory data-driven study generates new insights that confirm the uniqueness, and non-linearity of the relationship between goals and overall SDGs achievement.

Genotyping of Toxoplasma gondii Strains from Goats in Jahrom District, Southern Iran.

PURPOSE: Toxoplasma gondii is transmitted congenitally or acquired by consumption of food and water contaminated with cysts or oocysts. This study aimed at genotyping T. gondii strains from slaughtered goats in Jahrom. METHODS: A total of 561 specimens (heart, diaphragm, and tongue) from 187 slaughtered goats were collected from Jahrom slaughterhouse. After DNA extraction, the T. gondii strains were genotyped by the nested PCR-RFLP based on GRA6 and 3', and 5' ends of the SAG2 gene. RESULTS: T. gondii infection was present in 18.2% of cases. Among the examined organs, the diaphragm was more disposed to the infection (10.2%). Furthermore, infection rates of the heart and tongue were 8.6% and 3.7%, respectively. Concurrent infection in the heart and diaphragm, tongue and diaphragm, and heart and tongue were 3.2%, 0.5%, and 0.5%, respectively. In genotyping experiments, genotype I was the most frequent genotype of T. gondii (58.8%), followed by type II (23.5%), type III (11.8%), and a combination of type I and II (5.9%). CONCLUSIONS: The results of this study showed the presence of different genotypes of T. gondii in goats including three major and mixed genotypes. These results can be useful in toxoplasmosis control and prevention.

Bionomics of Phlebotomine Sand Flies in Different Climates of Leishmaniasis in Fars Province, Southern Iran.

Background: Phlebotomus and Sergentomya are distributed in the old-world regions and transmit leishmaniases through mammalian and reptile hosts. Cutaneous leishmaniasis (CL) is one of the most important diseases in Iran. Iranian sand flies belong to three Oriental, Palearctic, and Afrotropical Regions. Fars Province is located in Palearctic, but southern parts are affected Oriental region situations on phglebotomine population variety. Therefore, a comprehensive study was required on the vectors of the disease in this Province. Methods: Regarding the approved role of the environmental factors in creating the fauna and distribution of living creatures, the "de martonne climate method" was used, and the climate was noticed as an important environmental factor for the determination of vector distribution. Accordingly, 14 sampling sites were selected from 10 foci in different climates of Fars Province. 19648 sand flies were collected from the studied areas in this Province during 2016. Results: Phlebotomus papatasi and Se. antennata were the most frequent species, which were caught from in/outdoor areas. Phlebotomus sergenti and Ph. alexandri were caught from both Palearctic and Oriental zones but were more prevalent in the cold semi-arid climate of the Palearctic zone. Moreover, they were not caught from the hot desert and summer Mediterranean climates of the Oriental zone. Conclusions: It seems that Ph. papatasi as the main vector of CL could be well distributed in different climates in Fars. Moreover, some species like Ph. sergenti preferred especial climates in the Palearctic zone. Therefore, these data could be helpful to control leishmaniases more efficiently.

Molecular Detection of Leishmania Infection in Phlebotomine Sand Flies from an Endemic Focus of Zoonotic Cutaneous Leishmaniasis in Iran.

Background: Due to the outbreak of zoonotic cutaneous leishmaniasis (ZCL), a disease caused by Leishmania major and mainly transmitted by Phlebotomus papatasi, in Damghan City, Semnan Province, the probable vectors of the disease were investigated in the city from 20 March 2016 to 20 January 2018. Methods: Sand flies were collected from indoors and outdoors biweekly by sticky traps in different parts of the city. The trapped sand flies were stored in 70% ethanol. They were identified and checked for Leishmania infections using nested-PCR method and specific primers; CSB1XR, CSB2XF, LiR, and 13Z. Results: Overall, 1862 phlebotomine sand flies of Ph. papatasi (48.8%), Ph. andrejevi (8.3%), Ph. caucasicus (7.7), Ph. mongolensis (2%), Ph. sergenti (1.2%), Ph. alexandri (0.7%), Sergentomyia murgabiensis sintoni (29.3%), and Se. sumbarica (2%) were collected indoors (31.1%) and outdoors (68.9%). The highest and lowest numbers of collected sand flies were belonging to Ph. papatasi (48.8%) and Ph. alexandri (0.7%) respectively. 2.2% of the examined sand flies were shown to be infected with L. major and all were belonging to Ph. papatasi. Conclusion: This study confirms the report of Ph. papatasi infection with L. major and also the existence of Ph. sergenti and Ph. alexandri, the potential vectors of L. tropica and L. infantum respectively, in Damghan City. According to the findings, it is necessary for health officials to plan and take action to prevent the occurrence of ZCL epidemic in the city as well as the occurrence of other forms of leishmaniasis.

Frequency of pyrethroid resistance in human head louse treatment: systematic review and meta-analysis.

Head lice (Pediculus humanus capitis) are one of the most common insects causing infestations in humans worldwide, and infestation is associated with adverse socio-economic and public health effects. The development of genetic insensitivity (e.g., target site insensitivity = knockdown resistance or kdr) to topical insecticides has impaired effective treatment. Therefore, this study was undertaken to review and meta-analyze the frequency of pyrethroid resistance in treated head louse populations from the beginning of 2000 to the end of June 2021 worldwide. In order to accomplish this, all English language articles published over this period were extracted and reviewed. Statistical analyses of data were performed using fixed and random effect model tests in meta-analysis, Cochrane, meta-regression and I2 index. A total of 24 articles from an initial sample size of 5033 were accepted into this systematic review. The mean frequency of pyrethroid resistance was estimated to be 76.9%. In collected resistant lice, 64.4% were homozygote and 30.3% were heterozygote resistant. Globally, four countries (Australia, England, Israel, and Turkey) have 100% kdr gene frequencies, likely resulting in the ineffectiveness of pyrethrin- and pyrethroid-based pediculicides. The highest resistance recorded in these studies was against permethrin. This study shows that pyrethroid resistance is found at relatively high frequencies in many countries. As a result, treatment with current insecticides may not be effective and is likely the cause of increased levels of infestations. It is recommended that resistance status be evaluated prior to insecticide treatment, to increase efficacy.

TITLE: Fréquence de la résistance aux pyréthroïdes dans le traitement du pou de tête chez l'homme : revue systématique et méta-analyse. ABSTRACT: Les poux de tête (Pediculus humanus capitis) sont l'un des insectes les plus courants à l'origine d'infestations chez l'homme dans le monde, et l'infestation est associée à des effets socio-économiques et de santé publique néfastes. Le développement d'une insensibilité génétique (par exemple, l'insensibilité au site cible = résistance knockdown ou kdr) aux insecticides topiques a altéré l'efficacité de leur traitement. Par conséquent, cette étude a été entreprise pour examiner et méta-analyser la fréquence de la résistance aux pyréthroïdes dans les populations de poux de tête étudiées du début 2000 à la fin juin 2021 dans le monde. Pour ce faire, tous les articles en anglais publiés au cours de cette période ont été extraits et examinés. Les analyses statistiques des données ont été effectuées à l'aide de tests de modèles à effets fixes et aléatoires dans la méta-analyse, Cochrane, méta-régression et indice I2. Un total de 24 articles provenant d'un échantillon initial de 5033 ont été acceptés dans cette revue systématique. La fréquence moyenne de la résistance aux pyréthroïdes a été estimée à 76,9 %. Chez les poux résistants collectés, 64,4 % étaient homozygotes résistants et 30,3 % étaient hétérozygotes résistants. À l'échelle mondiale, quatre pays (Australie, Angleterre, Israël et Turquie) ont des fréquences de gène kdr de 100 %, ce qui entraîne probablement une inefficacité des pédiculicides à base de pyréthrine et de pyréthrinoïde. La résistance la plus élevée enregistrée dans ces études était celle contre la perméthrine. Cette étude montre que la résistance aux pyréthroïdes est trouvée à des fréquences relativement élevées dans de nombreux pays. En conséquence, le traitement avec les insecticides actuels peut ne pas être efficace et est probablement la cause d'une augmentation des niveaux d'infestation. Il est recommandé d'évaluer le statut de résistance avant le traitement insecticide, pour augmenter son efficacité.

In vitro evaluation of CRISPR PX-LmGP63 vector effect on pathogenicity of Leishmania major as a primary step to control leishmaniasis.

Cutaneous leishmaniasis (CL) is caused by intracellular obligate parasites (Leishmania spp.) carried by the blood-sucking of female sandflies and transmitted between mammalian hosts. Despite the high incidence and prevalence of Leishmania cases in many countries, it has been a neglected tropical disease. The current treatment approaches are limited by the complications such as loss of fertility and drug resistance. It is, therefore, essential to find new medicines to treat leishmaniasis. CRISPR/Cas9 as a powerful genome-editing tool provides the opportunity to create precise genetic manipulation to investigate the molecular basis of different leishmaniasis cases. Therefore, our main goal was to evaluate the CRISPR PX-LmGP63 vector effect on pathogenicity of Leishmania majorin vitroto challenge for using CRISPR/Cas9 as a therapeutic CL through the reduction of L. major pathogenicity by manipulating the GP63 gene. In this study, L. major parasites were transfected with CRISPR/Cas9 vectors constructed by electroporation and then added to macrophage cells on RPMI. The effect of CRISPR/Cas9 constructs on GP63 mutation, viability, and status of L. major was investigated by counting phagocytic parasites into macrophages and DNA sequence analysis. Our data validate that the use of CRISPR/Cas9 in L. major creates a new stop codon and disrupts the frame sheet of the gene by creating a new insertion (thymine), which prevents its expression. In addition, the parasite count was significantly different in the case and control of infected macrophages (P < 0.05). This study shows the successfully targeted manipulation of the L. major GP63 gene via the adaptation of the CRISPR/Cas9 editing tool. The manipulation of GP63 revealed a reduction in the infection load compared to wild-type parasite infection. Therefore, more studies are necessary for this field to help achieve a new method for the prevention and treatment of CL disease.

Genetic characterization of Toxoplasma gondii isolates from human spontaneous aborted fetuses in Jahrom, southern Iran.

Toxoplasma gondii (T. gondii) is an intracellular protozoan that infects the fetus through the placenta and leads to severe complications in the fetus. One of the complications of congenital toxoplasmosis is spontaneous abortion. The prevalence of toxoplasmosis infection was investigated among spontaneously aborted fetuses (SAFs), and the genotypes of parasite isolates were determined in the present study. Placentas from 330 samples of SAFs were collected in Jahrom (Fars province) from February to September 2018. DNA was extracted from each placental tissue. The T. gondii infection was detected using nested polymerase chain reaction (Nested-PCR) assay based on a 529 bp repeat element (RE) gene. Afterward, Toxoplasma was genotyped using PCR-restriction fragment length polymorphism (PCR-RFLP) based on the GRA6 gene. The frequency of T. gondii infection was found to be 14.5% (48 out of 330 samples). Genotyping of nine T. gondii isolates revealed that all belonged to genotype II. Statistically, the prevalence of T. gondii infection was significantly correlated with the education levels of the mothers and the age of the fetus (P < 0.05). The lowest prevalence of Toxoplasma infection belonged to mothers with university education and the highest frequency of infection was observed among the fetuses in the age group of 8-9 weeks. The findings of the present study suggest a significant role for toxoplasmosis in SAFs in Jahrom city.

Bionomics and phylo-molecular analysis of Leishmania species isolated from human lesions using ITS1 genes in north-east of Iran.

Leishmaniasis is a zoonotic infectious disease caused by Leishmania species. The identification of parasite species and the type of disease is beneficial for treatment and preventive modalities. Leishmania tropica and L. major have been reported as the main etiological agents of cutaneous leishmaniasis (CL) in Iran. The incidence of zoonotic CL has increased and different in distinct loci of Iran. Hence, we perused the Leishmania species and its genetic traits in the North East of Iran. The investigation was conducted on 200 positive smears prepared from patients' lesions suffering from CL referred to the health care centers of northeastern provinces in Iran from 2013 to 2019. The obtained positive microscopy samples were divided to score the ranges from + 1 to + 6, of them 40 smears exhibited low-parasitemia. Leishmania species analyzed using PCR-RFLP, genetic diversity indices evaluation, phylogenetic analysis, and sequencing comparison with other species in the GeneBank based on ITS1 gene. The isolated L. major strains were similar to other Iranian isolates in this region. Pairwise fixation index (FST) index was statistically significant in different L. major populations and showed the genetic differences in pairwise population of different geographical locations of Iran. The current study confirmed an old pattern endemicity of zoonotic CL in North-east of Iran. Therefore, in order to assess the hybrid formation, more epidemiological, ecological, and gene polymorphism studies are needed to understand the pathogenic role of Leishmania species in Iran.

Potential co-infection of Wolbachia with Leishmania among sand fly vectors caught from endemic leishmaniasis foci in Fars province, southern Iran.

Leishmaniasis is one of the Neglected Tropical Diseases in the tropical region of many countries in the world. The etiological agents (Leishmania parasites) of the disease are transmitted to human and other vertebrate hosts by infectious bites of female phlebotomine sand flies. On the other hand, some symbiotic microorganisms such as Wolbachia (Rickettsiales: Anaplasmataceae) may be transmitted vertically in many arthropods and may cause synergistic or antagonistic effects on epidemiology of the vector-borne diseases. Hence, in the present study, potential coinfection of Wolbachia with Leishmania in the sand fly vectors will be examined by PCR technique in the important leishmaniasis foci of Fars province in southern Iran, as a new feature for the disease long-term control. Sand flies were collected by sticky traps from indoor and outdoor locations of 5 different areas of Fars province during 2018 and 2019. DNAs of sand flies were extracted and PCR method was performed based on primers which were designed from surface proteins (WSP) genome region for Wolbachia and minicircle kDNA gene for Leishmania detections. At last, PCR products were sequenced and recorded in the GenBank. Out of 1002 sand flies caught from 5 different foci of Fars province, 909 male and female and 386 female sand flies' DNAs were extracted for detection of Wolbachia and Leishmania by PCR, respectively. Accordingly, out of the total 44 pools prepared from sand flies, 6 out of 28 pools form P. papatasi female were positive for Wolbachia in Shiraz, Marvdasht and Kharameh. However, none of female sand flies were positive for Leishmania. The study also focused on monitoring of sand flies co-infection of Wolbachia with Leishmania, which was not found in any of the studied samples. The negative results may be due to control strategies implemented which were done against Leishmaniasis in the studied areas during last years.

PCR-based detection of Toxoplasma gondii from cattle in southern Iran.

OBJECTIVE: Toxoplasma gondii is a protozoan parasite that is widely prevalent in most warm-blooded vertebrates. Humans mainly become infected by eating raw or undercooked meat. This study was designed to investigate the infection of cattle with T. gondii in Jahrom, southern Iran. METHODS: Tissue samples consisting of heart, diaphragm, and tongue were collected from 125 slaughtered cattle. DNA samples were extracted from the homogenized tissues. T. gondii was detected and genotyped using nested-polymerase chain reaction (Nested-PCR) and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) based on GRA6 and SAG2 (3', 5' terminal regions) genes, respectively. RESULTS: The prevalence of T. gondii DNA was 56% in cattle. The most infected tissue was the diaphragm (54.4%) followed by the heart (48.8%) and tongue (43.2%). Type II was the most prevalent genotype (70%) among T. gondii isolates. CONCLUSION: In this study, the high prevalence of T. gondii infection in cattle meat indicates the important role of cattle in the transmission of infection to humans. Therefore, incorporating the correct method of consuming meat in health education programs is crucial to prevent human infection.

High incidence of multidrug resistance and class 1 and 2 integrons in Escherichia coli isolated from broiler chickens in South of Iran.

The objective was to investigate the multidrug resistance and presence of class 1 and 2 integrons in 300 Escherichia coli isolates obtained from 20 broiler farms during three rearing periods (one-day-old chicks, thirty-day-old chickens, and one day before slaughter) in Fars, South Iran. Results showed that 81.00%, 82.00%, and 85.00% of isolates were multidrug-resistant on the first day, thirty-day-old chickens, and one day before slaughter, respectively. Multidrug-resistant E. coli isolates were further examined for the presence of class 1 and 2 integrons using PCR assay. The existence of class 1 integron-integrase gene (intI1) was confirmed in 68.40%, 72.70%, and 60.90% of multidrug-resistant isolates from stage 1, stage 2, and stage 3 of the rearing period, respectively. The frequency of class 2 integron-integrase gene (intI2) during the first to the third stage of sampling was 2.60%, 25.50%, and 30.40%. Also, sequence analysis of the cassette arrays within class 1 integron revealed the presence of the genes associated with resistance for trimethoprim (dfrA), streptomycin (aadA), erythromycin (ereA), and orfF genes. The results revealed that percentages of antimicrobial resistance in E. coli isolates were significantly higher in the middle and end stages of the rearing period. In conclusion, widespread dissemination of class 1 integrons in all three stages and rising trends of class 2 integrons existence in E. coli isolates during the rearing period of broiler chickens could exacerbate the spread of resistance factors among bacteria in the poultry industry. Future research is needed to clarify its implication for human health.