In Vitro Antileishmanial Activity of Achillea santolina Essential Oil against Leishmania infantum Promastigote by Methylthiazole Tetrazolium (MTT) and Trypan Blue Colorimetric Methods.

Leishmaniasis causes parasitic infections, especially in developing countries. The disease has not yet been controlled because of the absence of an effective vaccine and low-cost treatment. Achillea santolina essential oil (ASEO) might control the disease as it has antimicrobial properties. This study investigated the in vitro antileishmanial activity of ASEO against Leishmania infantum promastigote using the methylthiazole tetrazolium (MTT) and trypan blue colorimetric methods. The standard strain of L. infantum (MCAN/IR/96/LON49) promastigotes was prepared and cultured in a 96-well Novy-MacNeal-Nicolle (NNN) medium. The effects of different concentrations of saline, ASEO, and glucantime (10, 50, 100, 200, 500, and 1000 mg/mL) were examined in 24-, 48-, and 72-hour intervals using the MTT and trypan blue test methods.The use of ASEO reduced viability in all concentrations compared to the control group in times of 48 (p<0.05) and 72 h (p<0.05). Treatment with glucantime and ASEO had similar efficiency with the concentration of 1000 mL/mg in both methods after 72 h. The results showed that viability was significantly lower in the ASEO group with increases in time using both methods (p<0.05). Cohen's Kappa coefficient showed a significant agreement between the obtained results for the two methods (Kappa=0.856; p<0.001).In sum, the results showed in vitro antileishmanial activity of ASEO, but more clinical studies are needed to confirm the efficiency. ASEO can be used as an agent and/or in combination with synthetic agents for the treatment of leishmaniasis disease.

Detection of Fasciola Hepatica in Lori Sheep Using Polymerase Chain Reaction and Conventional Diagnostic Methods in Western Iran.

Fascioliasis is an emerging and important food and water-borne disease in human communities which has become one of the most important health challenges in countries, like Iran. It causes weight loss, a decrease in feed conversion ratio as well as milk and meat production, and also reduces fertility in animals the prevalence of fasciolosis is increasing in some regions of the world due to various factors. Different methods have been used for the detection of Fasciola hepatica in animals. This study is the first to detect F. hepatica in Lori sheep using polymerase chain reaction (PCR) and conventional diagnostic methods in Western Iran. During three months, 195 fecal samples were collected from sheep in Lorestan province, Iran, using the stratified random sampling method. The conventional diagnostic methods, including wet mount microscopic examination and concentration assays, as well as the PCR technique targeting the intergenic spacer gene of F. hepatica, were used for the detection of the parasite in sheep. In total, 4 (2.1%) out of 195 examined stool samples were positive for F. hepatica based on the conventional assays. The PCR test was positive for F. hepatica in7 (3.6%) samples of 195 studied specimens. Statistical analyses of the data revealed that there is a significant difference between the results of diagnostic methods for F. hepatica detection (P=0.0421). Finally, the results showed that PCR has more diagnostic sensitivity, compared to conventional diagnostic methods, including the concentration techniques and microscopic examination. Hence, it can be advised to use PCR for the detection of F. hepatica in sheep.

Serological Determination of Toxoplasma gondii among Sheep (Ovis aries) in Guilan Province, Iran.

Toxoplasma gondii is one of the most common foodborne protozoan parasite causing congenital infection, abortion, and stillbirth in humans and animals. The temperate and humid climate is one of the most important factors in the high prevalence of T. gondii. Sheep are among the important sources of meat production in Guilan province, Iran. Therefore, the consumption of raw and half-cooked meat is one of the major risk factors for T. gondii infection. Toxoplasmosis in patients with intact immune systems is usually asymptomatic; however, it but can be life-threatening in patients with a weak immune system (for example, patients with the human immunodeficiency viruses/acquired immunodeficiency syndrome or cancer and transplant recipients). Guilan is divided into three geographical regions of plains with a temperate climatic condition, hillsides with a semi-humid climate, and heights with cold mountainous climate. Climate situations play a role in the prevalence of toxoplasmosis. The present study aimed to investigate the seroprevalence of T. gondii infection among sheep in Guilan province, north of Iran. In the current cross-sectional study, a total of 400 sheep sera samples were tested for the determination of immunoglobulin G (IgG) antibody against T. gondii using the enzyme-linked immunosorbent assay. The samples were divided into different groups according to the geographical location and animal age. T. gondii antibody (i.e., IgG) was detected in 166 sheep (41.5%). The highest frequency of T. gondii infection (72.7%; n=56) was observed for the age group of &gt; 4 years; the difference was statistically significant in this regard (P=0.0001) in comparison to that reported for other groups. In addition, the seroprevalence of T. gondii was significantly higher in the plains (53.9%) than that of the hillsides and heights (P=0.0001). Consequently, the seroprevalence of T. gondii infection in Guilan was high indicating a significant relationship with geographical location and animal age.

Genetic Affinity of Echinococcus granulosus protoscolex in Human and Sheep in East Azerbaijan, Iran.

Echinococcosis caused by the larval form of Echinococcus granulosus (E. granulosus) is known as an important zoonotic disease in various parts of the world, including Iran. The genetic diversity of this parasite is very high, particularly in areas where the disease is endemic. It has been suggested in the literature from different parts of the world that diverse factors, such as parasite life cycle, transmission pathways, pathologic disease, immunization, and disease control can be affected by the genetic diversity of the parasite. Various studies indicated sheep strain G1 as the most common genotype throughout the world. This strain is commonly found in the liver and lung repeatedly causing echinococcosis in humans, sheep, and cattle. The present study was conducted to determine the genetic affinity between the protoscolex of E. granulosus in humans and sheep in East Azerbaijan province, Iran for the first time. A total of 120 hydatid cyst samples were collected, 60 of which were from people who referred to the hospitals of East Azerbaijan and 60 were from the sheep slaughtered in Tabriz slaughterhouse. Following DNA extraction, certain regions of the cox1 gene were amplified and evaluated by the polymerase chain reaction. The replicated parts in all isolates had the same size of 450 bp. Electrophoresis was followed by selecting a total of 60 suitable samples, including 30 human samples and 30 sheep samples and sending them for genome sequencing. The overlap of the samples was investigated using the BLAST software. The results of BLAST, sequencing, and overlap demonstrated a genetic linkage of approximately 91.76% between the protoscolex of E. granulosus in human and sheep.

Study on Cryptosporidium contamination in vegetable farms around Tehran.

In recent years, an increase in the number of cases of food-borne illnesses linked to fresh vegetables has been reported. One of the causative agents of these infections is Cryptosporidium and it appears that one route of transmission to humans is food-borne, so fruits and vegetables have important roles. The goal of this study was to determine the level of Cryptosporidium contamination in vegetable farms around Tehran, Iran. A total of 496 samples from 115 vegetable farms in different regions around Tehran (Capital city of Iran) were collected and different types of vegetables were investigated for the parasite in June and July, 2012. A sediment concentration method followed by modified Ziehl-Neelsen's acid-fast staining was used to determine the presence of Cryptosporidium oocysts. Our findings revealed that 6.6% of studied samples were contaminated with Cryptosporidium species. The highest rate of contamination was reported in Bagher Abad (South of Tehran) (11.1%), and green onions were more commonly contaminated (14.8%) than any other vegetables tested. Furthermore, when waste water was used to irrigate vegetable farms, the contamination rate was (33.3%). Statistical analysis showed a correlation between contamination with Cryptosporidium spp. and studied risk factors including: different regions around Tehran, type of vegetables, and type of water used for farm irrigation. Therefore, vegetables may provide a route by which Cryptosporidium can be transmitted to humans, and control strategies should be considered.

Study on Cryptosporidium contamination in vegetable farms around Tehran

In recent years, an increase in the number of cases of food-borne illnesses linked to fresh vegetables has been reported. One of the causative agents of these infections is Cryptosporidium and it appears that one route of transmission to humans is food-borne, so fruits and vegetables have important roles. The goal of this study was to determine the level of Cryptosporidium contamination in vegetable farms around Tehran, Iran. A total of 496 samples from 115 vegetable farms in different regions around Tehran (Capital city of Iran) were collected and different types of vegetables were investigated for the parasite in June and July, 2012. A sediment concentration method followed by modified Ziehl-Neelsen's acid-fast staining was used to determine the presence of Cryptosporidium oocysts. Our findings revealed that 6.6% of studied samples were contaminated with Cryptosporidium species. The highest rate of contamination was reported in Bagher Abad (South of Tehran) (11.1%), and green onions were more commonly contaminated (14.8%) than any other vegetables tested. Furthermore, when waste water was used to irrigate vegetable farms, the contamination rate was (33.3%). Statistical analysis showed a correlation between contamination with Cryptosporidium spp. and studied risk factors including: different regions around Tehran, type of vegetables, and type of water used for farm irrigation. Therefore, vegetables may provide a route by which Cryptosporidium can be transmitted to humans, and control strategies should be considered.

Cryptosporidiosis and its potential risk factors in children and calves in Babol, north of Iran.

This study was carried out during April-August 2009 to determine the prevalence of Cryptosporidium spp. and its potential risk factors in children and calves in Babol, north of Iran. A total of 150 faecal samples were taken directly from the rectums of calves which were no more than two months old. Information about age, breeding conditions, consistency of faeces (as diarrhoeic or normal) and contact with human were recorded. At the same time, 150 stool samples were taken from children aged one month to 6 years old in Amir Kola children hospital (Babol, north of Iran). All samples were stained with modified Ziehl-Neelsen's acid-fast and Auramine O techniques to detect for the presence of Cryptosporidium oocysts. Results revealed that the prevalence of Cryptosporidium in children and calves were 16% and 7.33% (Auramine O stain), and 10.67% and 4% (Modified Acid-fast stain), respectively. The prevalence of the infection according to age groups and consistency of faeces were found to be statistically significant with Auramine O stain. The prevalence of infection in urban and rural children was similar, but prevalence of Cryptosporidium was more in calves with native breeding. The young calves and children and type of animal breeding represent important risk factors for transmission of cryptosporidiosis. Moreover, there was no relationship between infection of Cryptosporidium in children and calves. Our finding revealed that clinical cryptosporidiosis cases exist in north of Iran and the most important infection route for Cryptosporidium spp. is anthroponotic transmission.

Epidemiological aspects of canine dirofilariasis in the north of iran.

BACKGROUND: Dirofilaria immitis is an important parasite in dog and other carnivores. Our objective was study on incidence and periodicity of heartworm in north of Iran and using other methods for its diagnosis in addition to Parasitology exam. METHODS: This survey spanned two years, between 2006 and 2008. Blood samples were collected from 431 stray dogs distributed along north of Iran, the coastal areas of the Caspian Sea. The Knott's modified test was used for diagnosis of D. immitis and other filariae. Meanwhile, the periodicity of microfilaria in peripheral blood circulation was calculated and the imaging diagnosis techniques of four dogs that had positive results were done. RESULTS: Diagnostic parasitology results indicated that 16.01% of stray dogs were microfilaremic. Two different microfilariae were diagnosed: D. immitis in 13.69%, Dipetalonema reconditum in 1.86% and in 0.46% both of them. There was no statistically significant between infection to fiariae with sex and age of dogs. Also study on the periodicity of the presence of microfilaria in peripheral circulation showed that the highest rate of those was at 1 am and the lowest rate at 12 pm. Radiographic study showed distinctive signs with varied degrees of severity included: Tortuous and enlargement of main and lobar pulmonary artery, pulmonary parenchymal lesions and Right side heart enlargement that confirmed in electrocardiography. Also in echocardiographic images observed short parallel-sided images with the appearance of equal signs that indicated the presence of the heartworm. CONCLUSION: These results showed that to obtain a reliable diagnosis of heartworm infection, imaging tests could support parasitological exams.