Molecular investigation of Leishmania in sandflies (Diptera: Psychodidae) and rodents (Mammalia: Rodentia) in Nahavand, west of Iran.

Cutaneous leishmaniasis caused by different species of Leishmania is transmitted by Phlebotominae sandflies. This disease remains a public health concern in Iran. Therefore, the present study aimed to examine Leishmania infection in sandflies and reservoir rodents in six rural regions of Nahavand, located in western Iran. From May to October 2022, sandflies and rodents were collected and identified at the species level. Additionally, rodents' skin lesions and earlobe specimens were collected separately for microscopic and molecular examination. All specimens were tested for Leishmania DNA by PCRs targeting the parasite's ITS-2 and 18S rRNA gene and positive were Sanger sequenced. A total of 3396 sandflies belonging to seven subgenera and 11 species, i.e., Phlebotomus papatasi (42.7%), P. major (20.6%), P. mascitti (0.3%), P. neglectus (0.2%), P. alexandri (0.2%), P. turanicus (0.3%), Sergentomyia murgabiensis (18.1%), S. dentata (10.5%), S. theodori (5.8%), S. antennata (1.1%), and S. pawlowski (0.1%) were identified. Based on the species population, 29 pools of sandflies were examined for the presence of Leishmania DNA using conventional PCR (cPCR), and individual DNAs were tested when positive. Leishmania major DNA was detected in two P. papatasi and Leishmania sp. in one P. major individual sandfly. This is the first report of Leishmania infection in sandflies from Hamadan province. The captured rodents (n = 61) belonged to four families and seven species, i.e., Arvicola amphibius (37.7%), Mus musculus (29.5%), Microtus socialis (13.1%), Apodemus sylvaticus (11.5%), Talpa davidiana (4.9%), Apodemus witherbyi (1.6%), and Rattus norvegicus (1.6%). Microscopic and molecular examinations of the rodent lesions and earlobes scored negative results. The presence of Leishmania in the Phlebotominae sandflies in Nahavand indicates a potential threat to humans and animals in the region. Regular monitoring and examination of the sandflies' population and timely diagnosis and treatment of new patients are strongly recommended.

The alleviating effects of sesame oil on diazinon-induced toxicity in male wistar rats.

Pesticides are introduced into the environment in a variety of ways and constitute a wide group of environmental pollutants. To evaluate the attenuating effects of sesame oil on the toxicity of diazinon (DZN), male Wistar rats were exposed to DZN and/or sesame oil by gavage at different dosages for 8 weeks. DZN in a concentration of 30 mg/kg caused an increase in the number of white blood cell (WBCs), and the combination of DZN and sesame oil raised the number of platelets; the number of red blood cells, however, did not change. In addition, DZN caused a drastic decrease in the sperm count in a dose-dependent manner and in a concentration of 50 mg/kg, the sperm count decreased by more than 50%, but the combination of sesame oil in a dose of 4 ml/kg with DZN reversed the effect of this pesticide. The evidence presented here suggests that in addition to antioxidants, such as olive oil, intermittent exposure with sufficient intervals can decrease the toxicity of pesticides.

Distribution and Seasonal Activity of Phlebotominae Sand Flies in Yazd and Its Outskirts, Center of Iran.

BACKGROUND: Phlebotominae sand flies are the main vectors of leishmaniasis and some other diseases. MATERIALS AND METHODS: Using sticky traps, sand flies were collected fortnightly from outdoors and indoors areas of selected sites. RESULTS: A total of 2032 specimens (498 in the city and 1534 in the outskirts of Yazd) belonging to 11 species were collected. The activity of sand flies started in early-April and ended in mid-November. There were two peaks of activity in the end of April and mid-September. Phlebotomus sergenti and P. papatasi were the most abundant species in the city and outskirts of Yazd city, respectively. Other species were P. salehi, P. ansarii, P. kazerouni, P. caucasicus, P. andrejevi, P. alexandri, P. mongolensis, Sergentomyia sintoni, and S. palestinensis. CONCLUSION: In comparison to some other parts of Iran, the extended period between two peaks suggests that the larvae to adult development of sand fly were delayed by the higher temperature of the summer months in Yazd province.

Bacterial Contamination of Collected Cockroaches and Determination Their Antibiotic Susceptibility in Khorramabad City, Iran.

Background: Cockroaches are one of the most important carriers of pathogenic microorganisms. Therefore, the presence of cockroaches in public places, especially in hospitals, homes, and restaurants, is dangerous, and threatens the health of society, people, and the environment. The aim of this study was evaluation of bacterial contamination of cockroaches and the sensitivity of these bacteria to various antibiotics, captured from Khorramabad City, Iran. Methods: This descriptive cross-sectional study was performed on 150 cockroaches collected from hospital environments, homes, and restaurants in Khorramabad. The outer surface of the cockroaches was washed with physiological saline. The suspension was centrifuged for 5 minutes at 2000rpm. Isolation and identification of bacteria was performed using phenotypic methods. Antibiotic susceptibility testing was performed by disk diffusion method according to Clinical and Laboratory Standard Institute (CLSI) guideline. Results: A total of 100 American cockroaches (66.66%), 28 B. germanica (18.66%) and 22 Blatta orientalis (14.66%) were identified. In total, 97.33% of the collected cockroaches were infected with bacteria. The most bacterial infection of the cockroaches was Escherichia coli, coagulase-negative Staphylococci and Bacillus respectively. The overall results of the antibiogram test showed that the identified bacteria were resistant to cephalothin, ampicillin, cefotaxime, and kanamycin antibiotics, semi-sensitive to ciprofloxacin and sensitive to tetracycline, gentamicin, nitrofurantoin, Trimethoprim/sulfamethoxazole, and Chloramphenicol. Conclusion: Infection of cockroaches with pathogenic bacterial agents in hospital, residential, and restaurant environments, as well as the observation of bacterial resistance to some common antibiotics is worrying.

Effects of vitamin A and vitamin E on attenuation of amphotericin B-induced side effects on kidney and liver of male Wistar rats.

Despite the fact that amphotericin B (AmB) is currently considered as the first choice for treatment of visceral leishmaniasis, it is associated with some side effects. This study was designed to investigate the protective effects of vitamins A and E against amphotericin B-induced adverse effects in the kidney and liver of rat. Thirty male Wistar rats aged 7-8 weeks and weighing around 200 g were randomly divided into five groups, each one containing six rats. The first to fifth groups received olive oil as the control groups, AmB, AmB + vitamin A, AmB + vitamin E, and AmB + vitamins A + E, respectively. Rats received vitamins by gavage (vitamin A, 1000 IU/kg and vitamin E, 100 IU/kg) and amphotericin B by injections (5.5 mg/kg body weight). The treatment was constantly continued for 5 days and days 7 and 21. At the end of the study, serum level of TAC, TOS, MDA, liver enzyme activity (ALT, AST, ALP, LDH), renal factors (urea, uric acid, and creatinine), lipid profile as well as histopathological changes of the liver and kidney were investigated. AmB significantly increased serum level of creatinine, urea, uric acid, ALP, TOS, MDA, and kidney and renal tissue damage (p < 0.05). Supplementation AmB with vitamins A and E alone or combination improved oxidative stress status, liver and renal tissue structure, and functional parameters and serum lipid profile. This study highlighted the effects of vitamin A and vitamin E on attenuation of amphotericin B-induced side effects on the kidney and liver of male Wistar rats. Combination of the two vitamins is more effective than either alone improving the oxidative stress status, serum lipid profile, or liver and renal tissue structure and functional parameters.

Control of Cockroaches (Blattaria) in Sewers: A Practical Approach Systematic Review.

Periplaneta americana (L.) (Blattaria: Blattidae), the American cockroach, is the most important invasive urban pest of sewer environments colonizing there with high significance of human public health and household allergens need to be controlled. Therefore this practical approach systematic review perform internationally to highlight and provide a detailed P. americana control in sewers. Of the 474 papers, 129 papers were selected to become this practical approach systematic review study of cockroach control in sewers. To control the American cockroaches, many studies have been conducted in various fields describing from an angle. The results were classified and discussed in getting cockroaches from sewers into buildings and their elimination, insecticide susceptibility, application of dust, bait and Inesfly paint insecticide formulations, biocontrol, and futuristic action categories. A recommending manner to achieve a successful P. americana cockroach control in sewers is using a combination of Integrated Pest Management (IPM) strategies resulted in significant reductions of cockroach infestations and asthma health outcomes. Use of P. americana breeding thelytoky, push-pull strategies and an automated sewer robot, and integrating health into the future buildings may be new approaches of P. americana control strategies.

Effect of seasonality on the population density of wetland aquatic insects: A case study of the Hawr Al Azim and Shadegan wetlands, Iran.

AIM: Wetlands are extremely suitable ecosystems to assess the effect of climate change on the density of aquatic insects. This study aimed to assess the effect of seasonality on populations of aquatic insects in the Hawr Al Azim and Shadegan wetlands. MATERIALS AND METHODS: The insect samplings were conducted at a large area of the Hawr Al Azim and five different sites of the Shadegan wetlands. In total, 18,534 arthropods of different life stages, including 12 orders containing 51 families, were collected and identified from the selected sites of the Shadegan and Hawr Al Azim wetlands. RESULTS: Results showed that the population density of wetland aquatic insects gradually increased as the average daily temperature decreased, positively increased with daily mean relative humidity and precipitation, and decreased with the mean daily evaporation between October and April. Conversely, the population density of wetland aquatic insects gradually decreased with increasing average daily temperature and reduction of the mean relative humidity and precipitation and increasing the average evaporation from April to September. When differences between the average daily and water temperatures reached minimum in April, the population density of wetland aquatic insects reached maximum and turned mainly to families that they have high level of biological indices, indicating that wetlands have clean waters around the spring. While around the autumn conversely, they mostly changed to families that they have low level of biological indices, indicating that wetlands have unclean waters. CONCLUSION: The present study showed an optimum condition for the growth of insects around spring. Seasonality affects the population density of wetland aquatic insects during a year.

10-Year Trends in Epidemiology, Diagnosis, and Treatment of Cutaneous Leishmaniasis in Hamadan Province, West of Iran (2007-2016).

Background: Cutaneous leishmaniasis is one of the most important infectious diseases in eastern Mediterranean countries. The aim of this study was to determine the epidemiological pattern of cutaneous leishmaniasis across a 10-year period in the Hamadan province. This study was considered necessary due to the lack of new information in recent years on the epidemiology of the disease. Methods: This is a descriptive study. The data of patients who were diagnosed with cutaneous leishmaniasis in Hamadan health centers were collected during 10 years (2007-2016) and were analyzed using SPSS software. Results: Of the 908 registered patients, 94% were male and 6% were female. The mean age was 32.7 ± 11.8. About 87.1% of patients had a history of travel to endemic areas. The highest incidence rate was observed in the year 2015 with a rate of 12.6, and the lowest was in 2008 at 1.5 per 100,000 people. Conclusion: Due to an increasing trend in the number of cutaneous leishmaniasis in Hamadan and the effect of occupation, high-risk groups such as lorry drivers should be informed of preventive measures such as using insect repellents. Also, considering the possibility of shaping a new hotspot in the province, thorough reviews and more comprehensive entomological studies are recommended.

The cytotoxicity of malathion and essential oil of Nepeta crispa (lamiales: lamiaceae) against vertebrate and invertebrate cell lines.

INTRODUCTION: Pesticides are used as essential tools to control vector-borne diseases and agricultural pests and maintain quality and quantity crop production. Scientists attempt to use derived plant natural products due to environmental safety and low mammalian toxicity. Therefore, the cytotoxicity of malathion and Nepeta crispa essential oil against vertebrate L929 and invertebrate Sf9 cell lines were investigated. METHODS: About 2×103 cells were placed into the wells of a 96-well plate experiments. Then appropriate concentrations of malathion and N. crispa essential oil added to the wells. The cells were allowed to grow for 3-5 days and estimated the cell numbers. Control cell wells contained only cells with DMSO. All treatments and controls repeated at least four replicates. RESULTS: About 2×103 cells were placed into the wells of a 96-well plate experiments. Then appropriate concentrations of malathion and N. crispa essential oil added to the wells. The cells were allowed to grow for 3-5 days and estimated the cell numbers. Control cell wells contained only cells with DMSO. All treatments and controls repeated at least four replicates. CONCLUSION: Plant essential oil not only had no negative effects but also had boosting effects on the L929 cell viability. Nepeta crispa essential oil had negative effects on the Sf9 cell viability with the differences that derived plant natural products containing environmentally friendly and readily biodegradable derivatives, hydrolyzing rapidly in nature and nearly having no destructive effects on mammals and environment.

Comparing cytotoxicity of propoxur and Nepeta crispa (Lamiales: Lamiaceae) essential oil against invertebrate (Sf9) and vertebrate (L929) cell lines.

BACKGROUND AND AIM: Attempts to use the plant products are to be an appropriate option due to substantial concerns about human health and environmental problems of using synthetic pesticides. Therefore, the cytotoxicity of Nepeta crispa essential oil was compared with propoxur against invertebrate (Sf9) and vertebrate (L929) cell lines. MATERIALS AND METHODS: The cell lines of Sf9 and L929 which were derived from the ovary glands of fall armyworm, Spodoptera frugiperda (Lepidoptera: Noctuidae) and mouse fibroblast cells, respectively, were obtained from the National Cell Bank of Pasteur Institute of Iran. About a number of 2 × 103 cells were placed into the wells of 96-well plate experiments. Then, appropriate concentrations of essential oil of N. crispa plant and propoxur added to the wells. The cells were allowed to grow for 3-5 days and estimated the numbers of cells. The cells of control experiment wells contained only cells with dimethyl sulfoxide. All control and treatment experiments repeated at least four replicates. RESULTS: Propoxur had negative effects on the viability of both invertebrate (Sf9) and vertebrate (L929) cell lines. The cytotoxicity of propoxur against invertebrate (Sf9) and vertebrate (L929) cell lines was gradually increased in accordance with propoxur concentrations. The cytotoxicity of N. crispa essential oil against vertebrate (L929) cell line was gradually decreased in accordance with plant concentrations, while the cytotoxicity of N. crispa essential oil against invertebrate (Sf9) cell line was strongly increased in accordance with plant concentrations. CONCLUSION: Plant essential oil not only had no negative effects but also had boosting effects on vertebrate cell viability. Essential oil of N. crispa plant had negative effects on invertebrate cell viability with the differences that the products derived from plants possessing of biodegradable and environmentally friendly derivatives, hydrolyzing rapidly in nature, and nearly having no destructive effects on environment, humans, or the mammals.

Hard tick (Acari: Ixodidae) species of livestock and their seasonal activity in Boyer-Ahmad and Dena cities of Kohgiluyeh and Boyer-Ahmad Province, Southwest of Iran.

AIM: The present study was carried out to identify the Ixodidae ticks fauna of livestock and their seasonal activity in the cities of Boyer-Ahmad and Dena of Kohgiluyeh Province, south-west of Iran. MATERIALS AND METHODS: Hard ticks from sheep, goats, and cattle were collected manually, stored in 70% ethanol, and identified using morphological characters. RESULTS: During the study, a total of 1273 hard ticks from four genera, including Rhipicephalus, Hyalomma, Dermacentor, and Haemaphysalis, were collected. Rhipicephalus sanguineus (s.l.) had the highest frequencies in both cities with 62.08 and 62.88% of collected specimens, followed by Hyalomma scupense with 14.36 and 13.54% in Boyer-Ahmad and Dena, respectively. Furthermore, Hyalomma marginatum with only one sample or 0.12% of collected ticks showed the lowest frequencies in the studied areas. Dermacentor marginatus with three samples or 0.37% was recorded only in Boyer-Ahmad, and Haemaphysalis sulcata with two samples or 0.43% was recorded only in Dena. In both cities, sheep were the most infested ruminant, and the ears in sheep and goats were the most affected areas. The highest activity was observed in spring, and the lowest activity was observed in winter and autumn. CONCLUSION: The results of the present study showed that Hyalomma and Rhipicephalus genera were the most widespread genera in the study areas. Regarding the importance of genera, such as Rhipicephalus, Hyalomma, and Haemaphysalis, in transmitting disease agents and the location of Kohgiloyeh and Boyer-Ahmad Province in the routes of migrant birds, further studies are required to elucidate their exact roles in human and livestock health in these areas.

Seasonal distribution and faunistic of ticks in the Alashtar county (Lorestan Province), Iran.

INTRODUCTION: Ticks are non-permanent obligate parasites that have considerable medical-veterinary and zoonosis importance. In this regard a study designed to investigate the distribution and fauna of ticks in the Alashtar county in Iran from April and March 2014. METHODS: Ticks were collected from livestock farms and facilities from selected rural and geographically location in the Alashtar county. Based morphological characteristics and reference identification keys, ticks were identified. RESULTS: A total of 549 ticks including 411 hard and 138 soft ticks were found. Ten tick species including Haemaphysalis concinna (0.36%), Haemaphysalis sulcata (0.36%), Hyalomma anatolicum (0.18%), Hyalomma dromedarii (0.18%), Hyalomma marginatum (1.45 %), Hyalomma schulzei (0.36%), Rhipicephalus annulatus (0.18%), Rhipicephalus bursa (28.1%), Rhipicephalus sanguineus (43.63%) and Argas persicus (25.2%) were identified. Tick seasonal distribution were 47.26%, 22.63%, 14.96% and 15.15% in the spring, summer, autumn and winter, respectively. The tick distribution was more from plain areas (64.96%) than the mountainous areas (35.04%). The rates of the tick contamination were 97.3% and 2.7% in the traditional and industrial livestock's, respectively. The livestock contamination ranks to the hard ticks were cattle (39.51%), sheep (34.15%) and goats (26.34 %), respectively. Chi-square analysis showed a significant difference among the seasonal distribution of the ticks in the spring, summer and autumn or winter; between the tick distribution in the plain and mountainous areas; and between the traditional and industrial livestock's tick contamination (P < 0.05). CONCLUSION: Present study proves to change the traditional livestock's to the industrial livestock's. These findings highlight the importance of ticks and shows need to their control and tick pest management.

Toxicity of zinc oxide nanoparticles on adult male Wistar rats.

The purpose of this study was to investigate the effects of zinc oxide nanoparticles (nZnO) on adult male Wistar rats. Thirty male Wistar rats divided into five groups of six animals each were used for this study. For ten days, Groups one to four continuously received 50, 100, 150 and 200 mg/kg nZnO, respectively. Group five served as the control group. At the end of the study, the rats were sacrificed and histopathological study of the liver and renal tissue, sperm analysis, serum oxidative stress parameters and some liver enzymes were done. The results of this study showed that nZnO at concentration more than 50 mg/kg lead to significant changes in liver enzymes, oxidative stress, liver and renal tissue and sperm quality and quantity. In conclusion, the toxicity of nZnO is more significant when the concentration is increased; however, the use of low doses requires further investigation.

An epidemiological study of cutaneous leishmaniasis using active case finding among elementary school students in Pakdasht, southeast of Tehran, Iran 2013-2014.

BACKGROUND: Cutaneous leishmaniasis is one of the prevalent health problems in tropical and subtropical areas including Iran. Therefore, it is essential to identify the prevalence and new foci of this disease in different regions of Iran. This study aimed to determine the epidemiology of cutaneous leishmaniasis by active surveillance in elementary schools in Pakdasht City, Southeast of Tehran, Iran 2013-2014. METHODS: This cross sectional study was carried out on 4800 students from 60 elementary schools in Pakdasht during October 2013 to March 2014. After examining the students, some epidemiological data such as age, gender, date, anatomic location of the lesion(s) in the body, number and size of lesion(s), lesion type, and travel history to endemic areas was recorded in a checklist. Students with active lesions were examined using parasitological and molecular methods (PCR). RESULTS: Totally, 31 students (0.64%) had leishmaniasis, of which 15 (0.31%, 95% CI, 0.15%, 0.47%) had active lesions and 16 (0.33%, 95% CI, 0.17%, 0.49%) had leishmaniasis scars. Molecular testing showed that Leishmania major was the causative agent of leishmaniasis in all patients with active lesion. The highest frequency of cutaneous leishmaniasis was found in the age group 10-12 years (P=0.03). There was a significant difference between the frequency of the disease in different months (P=0.04) and the anatomic location of the lesion/scar (P=0.04). Moreover, t-test revealed that there was a significant relationship between number of lesions/scars and the age group (P<0.001). CONCLUSIONS: The leishmanial type diagnosed in all patients was L. major. This finding suggests that Pakdasht might be a new focus for zoonotic cutaneous leishmaniasis.

Diversity and incrimination of sandflies (Psychodidae: Phlebotominae) captured in city and suburbs of Hamadan, Hamadan province, west of Iran.

OBJECTIVE: To investigate Phlebotomus sand fly fauna and their vectorial capacity in Hamadan, Iran. METHODS: During the three consecutive years (2010-2012), from May to October, sandflies were sampled monthly from residential areas of Hamadan and selected villages using sticky traps. RESULTS: A total of 3 089 sandflies comprising 12 species from 2 genera were collected, of which 6 species are known vectors of leishmaniasis in Iran and/or other parts of the world. About 75% of the collected sandflies were of two dominant species, Phlebotomus kandelaki (38%) and Phlebotomus papatasi (36.6%). These species in combination with Phlebotomus major (6.8%), Phlebotomus sergenti (6.3%), Phlebotomus caucasicus (5.3%), Phlebotomus wenyoni (1.3%), Sergentomyia sintoni (2.9%) and Sergentomyia dentate (2%) collectively comprised the majority of the sandflies. Other species including Phlebotomus halpensis, Phlebotomus longiductus, Phlebotomus comatus and Phlebotomus alexandri each comprised only less than 1% of collected samples. CONCLUSIONS: Although none of the 158 dissected female Phlebotomus kandelaki and Phlebotomus papatasi were infected with leishmania promastigote, it is probable that in the future, factors such as environmental changes, migration and urbanization can ease the transmission of leishmaniases in this area.