Co-detection and isolation of Leishmania and Crithidia among naturally infected Tatera indica (Rodentia: Muridae) in Fars province, southern Iran

Objective: To explore the co-detection of natural infection of Trypanosomatidae parasites such as Leishmania and Crithidia in reservoir hosts of leishmaniasis. Methods: Rodent populations were monitored in two endemic foci of cutaneous leishmaniasis of Fars province, southern Iran from March to October 2016. Rodents were trapped alive in several parts of Shiraz and Kharameh cities. Afterwards, their organs were prepared for detection of Leishmania and Crithidia species by molecular, microscopic, and culture methods. Results: Totally, 115 rodents of five species; Tatera indica (T. indica) (85), Rattus rattus (12), Meriones libycus (9), Mus musculus (7), and Rattus norvegicus (2), were trapped alive and their tissue samples were examined using microscopic, cultivation, and molecular assays. Overall, 59 (51.3%) rodents were positive for Leishmania or Crithidia parasites. The highest rate (61.2%; 52/85) of Leishmania infection was related to the T. indica population. The cultivation, and molecular observations showed that two (2.4%; 2/85) of T. indica (foot-pad, and spleen samples) were positive to Crithidia. Conclusions: This is the first report of Crithidia infection in T. indica in Iran. Consequently, more epidemiological and ecological studies are needed to understand the role of Crithidia and Leishmania in T. indica.

Synthesis and biological evaluation of metronidazole derivatives as anti-giardia agents

Nitroimidazole derivatives such as Metronidazole [MTZ] have been used as anti- protozoa and anti-anaerobic bacteria. In this study several compounds of MTZ derivatives were synthesized and evaluated against Giardia lamblia cyst. Firs MTZ were reacted with several alkyl halide to obtain O-alkyl MTZ derivatives, then products were purified and their chemical structures were confirmed by elemental and spectral analysis [1HNMR and Mass]. In order to biological evaluation all compounds were investigated against 25 Giardia samples isolated from giarfdiasis patients. Results showed compound 2a had the most activity on cyst of Giardia in comparison with MTZ

Knowledge, attitude, and practices related to cutaneous leishmaniasis in an endemic focus of cutaneous leishmaniasis, Southern Iran

Objective: To assess knowledge, attitudes, and practices (KAP) of inhabitants of an endemic area in Fars province toward cutaneous leishmaniasis (CL). Methods: The study was carried out in Lapui district in Fars province, south of Iran, one of the most important foci of CL in this province. Sample size (237 residents) was calculated based on population. House-to-house survey was done to collect the data regarding knowledge, attitudes, and practices of the inhabitants. The head of each household was interviewed by a trained staff to assess his/her KAP related to CL. A semi-structured KAP questionnaire was used for data collection.Results:Mean age of participants was 39 and more than half of the respondents were in the age group of 31-40. Males constituted 172 (72.5%) of subjects. Most of the respondents (84.3%) were literate. The majority of the study population (83%) had heard about Salak (local name for CL) and most of these respondents (91%) were aware that CL is presented with a cutaneous lesion. Nearly two-third of the participants (63.5%) stated the bite of mosquito (not specifically sandflies) for CL transmission. The respondents’ attitude regarding the treatment of CL was not satisfactory since only 48% believed that CL can be treated by medicine. A noticeable proportion of respondents (21%) believed in indigenous medicine for the treatment of CL. A small proportion of respondents (14%) stated that traditional healers are good at treating this disease. More than two-third (69%) of respondents believed that the disease is preventable although most of interviewees did not know about preventive measures. Conclusions: In this study, insufficient knowledge of community about infection nature, vector, transmission mode and preventive measures of CL, highlights the needs for a health education initiative to enhance the awareness of people about CL. This would improve inhabitants’ contribution in control program of CL in this area.

Laboratory based diagnosis of leishmaniasis in rodents as the reservoir hosts in southern Iran, 2012

Objective:To examine the fauna of rodents as zoonotic cutaneous leishmaniasis reservoir hosts in Zarqan County, Fars Province, south of Iran, during 2012. Methods:During 2012, wild rodents from different parts of this region were caught by Sherman traps and checked by the examination of liver and spleen smears, for Leishmania infection, to see which species were acting as reservoir hosts;the slides were then processed to extract DNA for molecular test using PCR assay. Results:From 108 rodent species caught, 63%were male and 37%identified as female. Meriones libycus was the most abundant species caught (80.5%) and 5.7%of them were found to be smear-positive for Leishmania amastigotes. The other species were Rattus rattus (14.8%) and Mus musculus (4.7%), but none of them were found positive. Leishmania infection was observed in male and female samples microscopically. Moreover, molecular results revealed Leishmania major in three male and two female specimens. Conclusions:Based on our knowledge, Meriones libycus is incriminated as the main reservoir hosts of Leishmania major in the rural area of Zarqan.

In vitro and in vivo potential of RH strain of toxoplasma gondii [type I] in tissue cyst forming

Based on recent studies, there are controversial reports on the capacity of tissue cyst forming of Toxoplasma gondii RH strain. In this study, the capacity was evaluated by in vivo and in vitro experiments. RH strain was subcutaneously inoculated to ten Wistar rats. After one month, their blood, brain, tongue and diaphragm were collected and evaluated by MAT, PCR, pathological and bioassay methods. The parasite was cultivated in the cell monolayer. To change to bradyzoite, the media pH was altered to 6.8. Biological aspect of the bradyzoites was evaluated by incubation in acidic pepsin and it's inoculation in ten BALB/c mice. All rats showed antibodies to Toxoplasma at titers >/= 1:320 but no DNA and tissue cyst were detected in the tissues. Following intraperitoneal inoculation of rats' brain homogenate into BALB/c mice, no infection was established in none of the animals. During presence of cell culture, in acid media for a 3-5 days period, cyst-like structures were noticed when they were stained with PAS. The visible bradyzoites in the cysts that were incubated in acid pepsin medium were not able to kill any mice. This study confirmed that Iranian RH strain has lost the potential of tissue cyst forming in rats and bradyzoites cultivated in cell culture lost their resistance to acidic condition, so this strain can be a candidate for future vaccine researches

Molecular genotyping of toxoplasma gondii in human spontaneous aborted fetuses in Shiraz, southern Iran

Congenital toxoplasmosis is associated with variable complications including encephalitis, microcephaly, hydrocephaly, hepatitis, lymphadenopathy and even intrauterine death. Presence of Toxoplasma gondii in human placenta may induce congenital infection. The aim of this study was to determine the genotypes of Toxoplasma gondii infection in human spontaneous aborted fetuses in Shiraz, south of Iran. Five hundred and forty two paraffin-embedded blocks of aborted placenta were collected, from two university-affiliated hospitals in Shiraz. Occurrence of spontaneous abortion was confirmed by examine of the slides. After re-cutting of the blocks and dewaxing, semi-nested PCR assay was used to detect the fragments of T. gondii B1 gene in the samples. Also direct molecular genotyping was performed on positive samples with Restriction Fragment Length Polymorphism-PCR analysis on the SAG2 gene. Among the 542 tissue samples, the B1 gene was amplified from 78 [14.4%] of cases with the semi nested PCR and typed by RFLP. The genotype of Toxoplasma strains of 65 [out of 78] PCR-positive samples were evaluated and 54 out of 65 [83.1%] were found to be type II and 11 out of 65 [16.9%] were type I. Considering the high level of Toxoplasma infection in aborted fetuses in this study, Toxoplasma might largely contribute to spontaneous abortion in this area of Iran

Report on leishmania infection of gerbillus nanus [rodentia: muridae] as the reservoir host of leishmania major in Hormozgan province

Cutaneous leishmaniasis or oriental sore has continuously expanded during the recent years in the country. Jask County in the easternmost point of Hormozgan province with 245 cases in 2007 has been the main focus of the disease. The present study was conducted to investigate the role of Gerbillus nanus as the reservoir of cutaneous leishmaniasis in this center. In a cross-sectional study during 2007-2008, rodents were caught from infected villages and after anesthesia, two slides were prepared from each ear of rodents using abrasive grinding and after recording morphometric specifications, their liver and spleen were kept in the 70% ethanol and the rest of the body was kept in 10% formalin for identification. Studies to determine leishmania infection were conducted through microscopic and molecular techniques. DNA was extracted through phenol/chloroform/lsoamyl alcohol method and it was proliferated through Nested-PCR method with primers LINR4, LIN17 and LIN19. A total of 106 rodents were caught. Species Gerbillus nanus [Muridae: Gerbillinae], with 17 heads, included 16.03% of haunting. Leishmania infection was found in a male sample of this species of rodent through microscopic method and two male and female samples [11.76%] through molecular method and specific PCR specified the parasite Leishmania major. Cutaneous leishmaniasis in this center is of zoonotic or damp type with leishmania major agent and therodentGerbillus nanus will be introduced in Hormozgan province for the first time as a possible reservoir host of the disease in this center. Infection of this species with Leishmaniamajoris reported for the first time in the world.

Leishmania major in Tatera indica in Estahban, Southern Iran: microscopy, culture, isoenzyme and PCR

This study identifies the infected rodent hosts to Leishmania major in Estahban town, southern Iran during 2004-2005. The rodents were caught alive from April 2004 to April 2005 in Estahban town, south of Iran and examined for any skin lesion. An impression was provided from the tissues of feet, tail, ears and any patent lesion, stained with Giemsa and studied microscopically for the presence of amastigotes. All samplings were cultured at 25°C in rabbit blood agar and considered negative if no promastigotes were visible during a two months period. The parasites from any positive culture were cryopreserved in liquid nitrogen pending their identification in PCR and isoenzyme electrophoresis. The femoral bones were histologically and ultrastrucrurally studied. Among 13 captured rodents, 8 were Tatera indica [5 male and 3 female Indian gerbils] and 5 were Rattus rattus [3 males and 2 females]. Just one female T. indica was smear-positive for amastigotes in Mohmmad Abad village. This rodent was also found culture positive for leishmanial infection which was confirmed by PCR and enzyme electrophoresis. At histological and ultrastructural levels, many clusters of amastigotes were noticed in the foamy macrophages of the femoral bone bone marrow. T. indica was found for the first time in the area and can be one of the rodents to be a potential reservoir host of L. major. It was also shown that femoral bone marrow was the tissue of choice to confirm the presence of macrophages containing the amastigote form of the parasite

Zoonotic cutaneous leishmaniasis in Shiraz, Southern Iran: a molecular, isoenzyme and morphologic approach

Cutaneous leishmaniasis [CL] with diverse clinical manifestations is prevalent and remains a major public health problem in Iran and its incidence has been doubled over the last decade. The present study is about the potential role of rodents in the epidemiology of CL in Kharameh district in Shiraz, Southern Iran. From April 2004 to April 2005, a total of sixteen rodents were collected in live traps from the endemic area of CL in Kharameh district in Shiraz. Evans medium was used for culture. Specific polymerase chain reaction and isoenzyme electrophoresis methods were performed to characterize the parasite. The rodent species were Tatera indica. Three samples from Tatera indica were found positive [2 males and 1 female in Kafdehak and Sejel-Abad villages] for L. major. Macrophages in the bone marrow of femoral bone were infected with the amastigote form of the parasite. It seems that T. indica is the reservoir host for CL in Kharameh [a district in Shiraz, Southern Iran]. It was shown that the bone marrow of the rodents is the tissue of choice for light and ultrastructural studies of L. major