Molecular and haematological investigations on Anaplasma ovis infection in goats in Ahvaz, Iran: Insights into infection rate, haemolytic effect and breed influences.

BACKGROUND: Anaplasma ovis (A. ovis) is the predominant causative agent of anaplasmosis in goats and sheep in most tropical and subtropical regions of the world. However, there is considerable variation in reported infection rates, breed susceptibility, and controversial findings regarding the haemolytic effects of A. ovis infection in goats. OBJECTIVES: Thus, we investigated the molecular and haematological aspects of A. ovis infection in goats from Ahvaz city. METHODS: One hundred and fifty apparently healthy goats (74 blacks and 76 Najdi goats) were randomly sampled from six flocks in the Ahvaz suburb during ticks' activity season. Haematological evaluation, smear microscopic (SM) examination and PCR assay were performed to assess A. ovis infection. Additionally, the percentage of parasitemia was determined from blood smears. RESULTS: SM examination revealed that 25.7% of the goats displayed erythrocyte Anaplasma-like inclusion bodies. PCR analysis indicated that 54% of the goats were positive for A. ovis infection (44.6% of blacks and 63.2% of Najdi goats). No significant difference in haematological values was observed between healthy and infected goats based on PCR testing. However, a significant difference in haematological indices was observed between the group with parasitemia level of 0.01-0.02% (SM and PCR positive) compared to the healthy goats (SM and PCR negative), particularly concerning Hb, PCV and RBC count (p < 0.01). CONCLUSIONS: When the parasitemia exceeds 0.01%, A. ovis infection may disrupt haematological parameters in infected goats. The high prevalence of A. ovis infection (54%) among the studied goats underscores the importance of giving special attention to implementing necessary measures for disease control in the Ahvaz suburb.

Isoenzyme characterization of Leishmania infantum toward checking the antioxidant activity of superoxide dismutase and glutathione peroxidase.

BACKGROUND: Leishmania infantum is the major causative agent of visceral leishmaniasis in Mediterranean regions. Isoenzyme electrophoresis (IE), as a biochemical technique, is applied in the characterization of Leishmania species. The current study attempted to investigate the isoenzyme patterns of logarithmic and stationary promastigotes and axenic amastigotes (amastigote-like) of L. infantum using IE. The antioxidant activity of superoxide dismutase (SOD) and glutathione peroxidase (GPX) was also checked in the aforementioned forms. METHOD: After L. infantum cultivation and obtaining logarithmic and stationary promastigotes, axenic amastigotes were achieved by incubation of stationary promastigotes at 37 °C for 48 h. The lysate samples were prepared and examined for six enzymatic systems including glucose-6-phosphate dehydrogenase (G6PD), nucleoside hydrolase 1 (NH1), malate dehydrogenase (MDH), glucose-phosphate isomerase (GPI), malic enzyme (ME), and phosphoglucomutase (PGM). Additionally, the antioxidant activity of SOD and GPX was measured. RESULTS: GPI, MDH, NH1, and G6PD enzymatic systems represented different patterns in logarithmic and stationary promastigotes and axenic amastigotes of L. infantum. PGM and ME showed similar patterns in the aforementioned forms of parasite. The highest level of SOD activity was determined in the axenic amastigote form and GPX activity was not detected in different forms of L. infantum. CONCLUSION: The characterization of leishmanial-isoenzyme patterns and the measurement of antioxidant activity of crucial antioxidant enzymes, including SOD and GPX, might reveal more information in the biology, pathogenicity, and metabolic pathways of Leishmania parasites and consequently drive to designing novel therapeutic strategies in leishmaniasis treatment.

Disentangling detrimental sand fly-mite interactions in a closed laboratory sand fly colony: implications for vector-borne disease studies and guidelines for overcoming severe mite infestations.

BACKGROUND: Vector sand fly colonies are a critical component of studies aimed at improving the understanding of the neglected tropical disease leishmaniasis and alleviating its global impact. However, among laboratory-colonized arthropod vectors of infectious diseases, the labor-intensive nature of sand fly rearing coupled with the low number of colonies worldwide has generally discouraged the widespread use of sand flies in laboratory settings. Among the different factors associated with the low productivity of sand fly colonies, mite infestations are a significant factor. Sand fly colonies are prone to infestation by mites, and the physical interactions between sand flies and mites and metabolites have a negative impact on sand fly larval development. METHODS: Mites were collected from sand fly larval rearing pots and morphologically identified using taxonomic keys. Upon identification, they were photographed with a scanning electron microscope. Several mite control measures were adopted in two different laboratories, one at the Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases-National Institutes of Health (Rockville, MD, USA), and the other at the University of Calgary (Calgary, AB, Canada). RESULTS: The mite species associated with sand fly colonies in the two laboratories were morphologically identified as Tyrophagus sp. and Stratiolaelaps scimitus. While complete eradication of mites in sand fly colonies is considered unrealistic, drastically reducing their population has been associated with higher sand fly productivity. CONCLUSIONS: We report a case of detrimental interaction between sand flies and Tyrophagus sp. and S. scimitus in a closed laboratory sand fly colony, discuss their impact on sand fly production and provide guidelines for limiting the mite population size in a closed laboratory colony leading to improved sand fly yields.

First Report of Isolation and Characterization of Acanthamoeba spp. from the Milk Used for Calf Feeding.

PURPOSE: Acanthamoeba spp. can be found in natural and artificial environments, which reflects their high adaptability to different conditions. Based on the available data, there is scarce information about the isolation of amoeba from milk. This study aimed to investigate the probable presence of Acanthamoeba in milk used for calf feeding. METHODS: 200 milk samples from 50 industrial and traditional farms were collected. The samples were filtered and cultured on the 1.5% Non-nutrient agar medium. The amoebic growth was examined with an inverted microscope daily. DNA was extracted from the positive plates, and a PCR reaction was undertaken using the primers amplifying the Acanthamoeba 18 S rRNA gene. Five samples were purified and sequenced using specific primers. Maximum likelihood reconstructions were performed using the phylogenetic program MEGA software. The osmo and thermotolerance of isolated trophozoites were examined as well. RESULTS: Out of 200 milk samples, Acanthamoeba was isolated from 27 (13.5%). The phylogenetic tree represents that all the isolates belonged to the genotype T4. Results of thermo and osmotolerance tests showed that isolates could develop at 37 and 43 ◦C. Besides, trophozoites survived at 0.5 M mannitol and 1 M. CONCLUSION: For the first time, Acanthamoeba spp. were isolated from milk used to feed dairy calves. Due to Acanthamoeba's neglected role in pathogen persistence and survival, hygiene instructions should be reconsidered.

Sublethal Exposure to Plasma-Activated Water Influences the Morphological Characteristics, Phagocytic Ability, and Virulence of Acanthamoeba castellanii.

PURPOSE: This study aimed to examine the ultrastructure, cytotoxicity, phagocytosis, and antioxidant responses of Acanthamoeba castellanii trophozoites exposed to sublethal plasma-activated water. METHODS: Trophozoites were exposed to a sublethal treatment of PAW and compared to untreated viable trophozoites via adhesion assays on macrophage monolayers, osmo- and thermotolerance tests. Bacterial uptake was assessed in treated cells to evaluate their phagocytic characteristics. Oxidative stress biomarkers and antioxidant activities were compared in treated and untreated trophozoites. Finally, the expression of the mannose-binding protein (MBP), cysteine protease 3 (CP3), and serine endopeptidase (SEP) genes was determined in cells. RESULTS: In PAW-treated trophozoites, cytopathic effects were more extensive and resulted in the detachment of macrophage monolayers. Treated trophozoites could not grow at high temperatures (43 °C). Moreover, they showed osmotolerance to 0.5 M D-mannitol but not to 1 M. Results demonstrated a higher bacterial uptake rate by PAW-treated trophozoites than untreated cells. Activities of superoxide dismutase and catalase and catalase were significantly greater in the treated trophozoites, and the glutathione and glutathione/glutathione disulfide were significantly lower in the PAW-treated cells. Exposure to PAW also significantly increased the malondialdehyde level and total antioxidant capacity. Treatment with PAW led to significantly higher expression of virulent genes like MBP, CP3, and SEP. CONCLUSION: PAW is a double-edged sword against A. castellanii. PAW is an effective antiamoebic agent in proper usage, whereas its sublethal exposure may reduce its effectiveness and increase amoebas' pathogenicity. An agent's adequate concentration and exposure time are essential to achieve optimum results.

Exposure to sublethal concentrations of chlorine enhances the cytotoxicity of Acanthamoeba castellanii.

Free-living amoebae belonging to the genus Acanthamoeba are the causative agents of infections in humans and animals. Many studies are being conducted to find effective compounds against amoebae, but their sublethal concentration effects on surviving amoebae seem to have been overlooked. Chlorine is a common disinfection agent commonly added to public water facilities and supplies. In this study, the cytopathic and phagocytic properties of Acanthamoeba castellanii trophozoites following exposure to sublethal concentrations of chlorine were examined. Two hours of exposure to 5 ppm hypochlorite calcium was considered the sublethal concentration for A. castellanii trophozoites. To compare the pathogenic potential of treated and untreated Acanthamoeba trophozoites, cytotoxicity, adhesion assays in RAW 264.7 macrophages, osmo, and thermotolerance tests were carried out. Bacterial uptake was assessed in treated cells to evaluate their phagocytic characteristics. Oxidative stress biomarkers and antioxidant activities were compared in treated and untreated trophozoites. Finally, the mRNA expression of the mannose-binding protein (MBP), cysteine protease 3 (CP3), and serine endopeptidase (SEP) genes was determined in cells. In all the experiments, untreated trophozoites were considered the control. In comparison to untreated trophozoites, in chlorine-treated trophozoites, cytopathic effects were more extensive and resulted in the detachment of macrophage monolayers. Treated trophozoites could not grow at high temperatures (43 °C). Besides, they showed osmotolerance to 0.5 M D-mannitol but not to 1 M. Results demonstrated a higher bacterial uptake rate by chlorine-treated trophozoites than untreated cells. The treated and untreated cells had significantly different glutathione and glutathione/glutathione disulfide ratios. Antioxidant enzyme activities, total antioxidant capacity, and malondialdehyde levels were increased significantly in chlorine-treated cells. Quantifying mRNA expression in chlorine-treated trophozoites revealed that virulence genes were upregulated. Chlorine can form resistance and virulent amoebae if it is not used at a proper concentration and exposure time. Identification of stress responses, their mechanisms in Acanthamoeba, and their relation to amoeba virulence would give us a better perception of their pathophysiology.

Evaluation of hematological changes, oxidant/antioxidant status and immuno-logical responses in sheep and goats naturally infected with Linguatula serrata.

Linguatula serrata is a worldwide zoonotic food-borne parasite. The parasite is responsible for linguatulosis and poses a concern to human and animal health in endemic regions. This study investigated the hematological changes, oxidant/antioxidant status and immunological responses in goats and sheep naturally infected with L. serrata. Hematological changes, antioxidant enzymes and malondialdehyde (MDA) levels were measured. The level of inter-leukin-2 (IL-2), IL-4, IL-5, IL-10, and tumor necrosis factor alpha (TNF-α) mRNA expression was investigated in lymph nodes. According to the hemogram results, eosinophils were significantly increased in the infected goats and sheep, and Horizontal Gene Transfer (HGT), hematocrit (HCT), and mean corpuscular hemoglobin concentration (MCHC) were significantly decreased. The levels of MDA and the activity of glutathione peroxidase (GPx) were significantly higher in infected animals than in non-infected animals. However, the activity of superoxide dismutase (SOD) and catalase (CAT) was significantly lower in infected animals than in non-infected animals. A comparison of the cytokine mRNA expression in lymph nodes from infected and non-infected animals showed higher cytokine expression in the infected animals. Infection with L. serrata caused microcytic hypochromic and normocytic hypochromic anemia in goats and sheep. The inconsistent results of immunological changes were found in infected goats and sheep. In both animals, oxidative stress occurred and led to an increase in lipid peroxidation. L. serrata created a cytokine microenvironment biased towards the type 2 immune responses.

Efficacy of light chain 3-fused protein multi epitope in protection of mice challenged with Mycobacterium tuberculosis.

The new strategy for vaccine development such as the fused protein multi-epitope capable of preventing the reactivation of latent tuberculosis infection (LTBi) can be an effective strategy for controlling tuberculosis (TB) worldwide. This study was conducted to evaluate the immunity of experimentally infected BALB/c mice with Mycobacterium tuberculosis after injection of DNA construct. Nineteen female BALB/c mice were divided into three groups and injected with 0.50 mL of M. tuberculosis. After 3 weeks, lung and spleen samples from the infected mice were examined. The protective effects of light chain 3-fused protein multi-epitope against TB were evaluated for post-exposure and therapeutic exposure. The lungs and spleens of the mice were aseptically removed after death for histopathology analysis. The bacterial colonies were counted, and the cells were stained after 3 weeks of incubation. No significant differences were observed between the post-exposure and therapeutic exposure groups. The pathological changes in the lung tissue of mice in these groups included an increase in the thickness of interalveolar septa, hyperemia, and intraparenchymal pulmonary hemorrhage centers (positive control), scattered hyperemic areas (negative control), and hyperemia in the interstitial tissue, scattered hyperemic areas in the lung parenchyma and lymphocytic infiltration centers (experimental group). Flow cytometry of the post-exposure and therapeutic exposure models showed insignificant changes in all three groups. It seems necessary to develop a post-exposure and therapeutic exposure vaccine strategy that focuses on LTBi to prevent the progression of the active disease. In this regard, multi-epitope vaccines should be designed to induce both cellular and humoral immunity.

Biofilm formation of Salmonella enterica serovar Enteritidis cocultured with Acanthamoeba castellanii responds to nutrient availability / La formación de biopelículas de Salmonella enterica serovar Enteritidis cocultivada con Acanthamoeba castellanii responde a la disponibilidad de nutrientes

Acanthamoeba spp. and Salmonella share common habitats, and their interaction may influence the biofilm-forming ability of Salmonella. In this study, biofilm formation of Salmonella enterica serovar Enteritidis cocultured with Acanthamoeba castellanii was examined in nutrient-rich and nutrient-deficient media. Furthermore, transcript copy number of biofilm-related genes in the biofilm cells of S. Enteritidis in monoculture was compared to those in coculture with A. castellanii. Results demonstrated that the presence of A. castellanii in the culture media activates the genes involved in the biofilm formation of S. Enteritidis, regardless of the nutrient availability. However, biofilm formation of S. Enteritidis cocultured with A. castellanii was not consistent with the transcript copy number results. In nutrient-rich medium, the number of Salmonella biofilm cells and the contents of the three main components of the biofilms including eDNA, protein, and carbohydrates were higher in the presence of A. castellanii compared to monocultures. However, in nutrient-deficient medium, the number of biofilm cells, and the amount of biofilm components in coculture conditions were less than the monocultures. These results indicate that despite activation of relevant genes in both nutrient-rich and nutrient-deficient media, biofilm formation of S. Enteritidis cocultured with A. castellanii responds to nutrient availability.(AU)

Levels of motivation and basic psychological need satisfaction in nursing students: In perspective of self-determination theory.

BACKGROUND: Motivation is the first and most important constituent element of learning behavior. One of the most important theories in this field is self-determination theory (SDT) which is a general theory of motivation. According to this theory, the satisfaction of basic psychological needs of autonomy, competence, and relatedness are effective in the internalization of incentive. Hence this theory can be a good foundation for reforming medical education programs. OBJECTIVE: The aim of this study was determining the basic psychological need and the effect of it on level of motivation and self-determined motivation of nursing student in the field. METHODS: This is a cross-sectional, descriptive-analytical study done through a census on 243 nursing students of Guilan University of Medical Sciences. Data gathering tool was demographic information, motivation level, and basic psychological needs questionnaire. Data were analyzed using independent t-test, Spearman and Pearson correlation, Man-Whitney, backward regression considering P < 0.05. RESULTS: The findings showed that the majority of students (51.9 %) were female and, the mean age was 21 years. Statistical tests indicated a significant relationship between levels of motivation, basic psychological needs, and demographic variables. However, the regression coefficients indicated that the need for competence and relatedness could be a suitable predictor for internal motivation. CONCLUSION: Basic psychological needs satisfaction, especially needs of competence and relatedness in the clinical field by instructors can lead to internalization of their incentive and positive outcomes.

Biofilm formation of Salmonella enterica serovar Enteritidis cocultured with Acanthamoeba castellanii responds to nutrient availability.

Acanthamoeba spp. and Salmonella share common habitats, and their interaction may influence the biofilm-forming ability of Salmonella. In this study, biofilm formation of Salmonella enterica serovar Enteritidis cocultured with Acanthamoeba castellanii was examined in nutrient-rich and nutrient-deficient media. Furthermore, transcript copy number of biofilm-related genes in the biofilm cells of S. Enteritidis in monoculture was compared to those in coculture with A. castellanii. Results demonstrated that the presence of A. castellanii in the culture media activates the genes involved in the biofilm formation of S. Enteritidis, regardless of the nutrient availability. However, biofilm formation of S. Enteritidis cocultured with A. castellanii was not consistent with the transcript copy number results. In nutrient-rich medium, the number of Salmonella biofilm cells and the contents of the three main components of the biofilms including eDNA, protein, and carbohydrates were higher in the presence of A. castellanii compared to monocultures. However, in nutrient-deficient medium, the number of biofilm cells, and the amount of biofilm components in coculture conditions were less than the monocultures. These results indicate that despite activation of relevant genes in both nutrient-rich and nutrient-deficient media, biofilm formation of S. Enteritidis cocultured with A. castellanii responds to nutrient availability.

Chlorine-stressed Salmonella cells are significantly more engulfed by Acanthamoeba trophozoites and have a longer intracystic survival than the non-stressed cells.

In the present study, the effect of sublethal chlorine-induced oxidative stress on the subsequent interaction of Salmonella enterica serovars Enteritidis and Typhimurium with Acanthamoeba castellanii and A. polyphaga was evaluated. Sublethal chlorine concentration was determined using the lag phase extension information and used to prepare chlorine-stressed Salmonella cells. Coculture experiments of Acanthamoeba and Salmonella cells were performed in Page's amoeba saline (PAS) at 25 °C for 2 h. The results showed that the chlorine-stressed Salmonella cells were significantly more engulfed by A. castellanii and A. polyphaga trophozoites than the non-stressed cells. The uptake rates of the chlorine-stressed and non-stressed Salmonella cells were in the range of 14.17-27.34 and 6.51-11.52% for A. castellanii, and in the range of 8.32-17.76 and 2.28-6.12% for A. polyphaga trophozoites, respectively. Moreover, intracystic survival time of chlorine-stressed cells of S. Enteritidis and S. Typhimurium was significantly longer than that of non-stressed cells. While, non-stressed Salmonella cells survived within A. castellanii and A. polyphaga cysts for 13-20 and 8-15 days, chlorine-stressed cells were recovered from A. castellanii and A. polyphaga cysts after 22-32 and 15-24 days, respectively. These results underscore the importance of bacterial exposure to sublethal chlorine concentrations in their interaction with free-living amoebae, and may lead to a better understanding of the parameters affecting the persistence of Salmonella enterica serovars in food-related environments.

Hematological, immunological, and polyamines alterations in the concomitant occurrence of Fasciola gigantica and hepatic leiomyoma in cattle.

No previous studies have investigated the polyamines alterations during fascioliasis due to F. gigantica in ruminants. This study was therefore carried out to find out the possible relationship between the extent of liver destruction and leiomyoma and some hematological and immunological parameters and polyamines alterations in F. gigantica infection. Fifty cattle with liver fascioliasis and fifteen healthy cattle were selected for the study. For the histopathological study, liver tissue samples were stained with hematoxylin and eosin (H&E) and Masson's Trichrome methods. The leiomyoma suspected specimens were immunohistochemically stained for smooth muscle actin and desmin. Different hematological parameters were investigated in infected and non-infected animals. Furthermore, levels of putrescine, spermidine, and spermine were measured in homogenized liver samples. Serum IL-4 and TNF-α levels were also evaluated. By histological examination, the lesions were noted in all the infected specimens. These lesions were varied from leiomyoma, chronic catarrhal cholangitis, arteriosclerosis, telangiectasia, and fresh migratory tunnels filled with RBC and eosinophils. Comparison of hemogram results between infected and non-infected groups revealed a significant decrease in red blood cell counts (RBC), mean corpuscular hemoglobin concentration (MCHC), and platelet count (PLT) in infected animals. Also, a significant elevation in mean corpuscular volume (MCV) concentration was detected in infected animals. The putrescine and spermine levels of the infected animals were significantly higher than the non-infected animals. Although spermidine was increased in infected livers, its elevation was not significant. Based on the results, the level of IL-4 and TNF-α was not significantly changed in infected animals. In conclusion, the concurrent occurrence of leiomyoma and fascioliasis due to F. gigantica and polyamines elevation (putrescine and spermine) is reported for the first time. The role of polyamines in the concurrent occurrence of leiomyoma and fascioliasis is an area for future research.

An Enzyme-Linked Immunosorbent Assay for the Detection of Antibodies to Linguatula serrata in Experimentally Infected Dogs.

BACKGROUND: Linguatula serrata is a causative agent of visceral and nasopharyngeal linguatulosis in humans and animals. The aim of the present study was to investigate the immune response of dogs experimentally infected by L. serrata with ELISA. METHODS: Five puppies were infected by inserting the L. serrata nymphs in their nasal cavities (infected group) in the Department of Parasitology of Shahid Chamran University of Ahvaz, during 2018-2019. Three animals were kept as the non-infected control group. Blood samples were collected from the animals for seven months at approximately monthly intervals for serum preparation. Nasal samples were taken weekly from the fourth month. ELISA was designed and performed on 64 sera (24 negatives, and 40 positives) using somatic (S), and excretory-secretory (ES) antigens. RESULTS: Overall, 100% of the animals were infected with the parasite. Based on the results of ELISA, the ES antigen (sensitivity 95% and specificity 92%) was more preferred than the S antigen (sensitivity 95% and specificity 85%). Female parasites had significant effects on the immune response. There was a significant correlation between the clinical symptoms and the presence of female parasites (P<0.05). CONCLUSION: The results showed a practical method for dogs' experimental infection. ELISA method is suitable for the detection of infection at different stages of development, especially before the maturation stage of the parasite. In this regard, the ES antigen of the parasite was more immunogenic. Therefore, ELISA can be used as a serological method in the early detection and epidemiological studies of infection with L. serrata in dogs.

Evaluating mechanism and severity of injuries among trauma patients admitted to Sina Hospital, the National Trauma Registry of Iran.

PURPOSE: Injuries are one of the leading causes of death and lead to a high social and financial burden. Injury patterns can vary significantly among different age groups and body regions. This study aimed to evaluate the relationship between mechanism of injury, patient comorbidities and severity of injuries. METHODS: The study included trauma patients from July 2016 to June 2018, who were admitted to Sina Hospital, Tehran, Iran. The inclusion criteria were all injured patients who had at least one of the following: hospital length of stay more than 24 h, death in hospital, and transfer from the intensive care unit of another hospital. Data collection was performed using the National Trauma Registry of Iran minimum dataset. RESULTS: The most common injury mechanism was road traffic injuries (49.0%), followed by falls (25.5%). The mean age of those who fell was significantly higher in comparison with other mechanisms (p < 0.001). Severe extremity injuries occurred more often in the fall group than in the vehicle collision group (69.0% vs. 43.5%, p < 0.001). Moreover, cases of severe multiple trauma were higher amongst vehicle collisions than injuries caused by falls (27.8% vs. 12.9%, p = 0.003). CONCLUSION: Comparing falls with motor vehicle collisions, patients who fell were older and sustained more extremity injuries. Patients injured by motor vehicle collision were more likely to have sustained multiple trauma than those presenting with falls. Recognition of the relationship between mechanisms and consequences of injuries may lead to more effective interventions.

The potential therapeutic effect of adipose-derived mesenchymal stem cells in the treatment of cutaneous leishmaniasis caused by L. major in BALB/c mice.

Leishmaniasis is one of the most neglected tropical infectious diseases in the world. The emergence of drug resistance and toxicity and the high cost of the available drugs with a lack of new anti-leishmanial drugs highlight the need to search for newer therapies with anti-leishmanial activities. Due to the mesenchymal stem cell (MSC) immunomodulatory capacity, they have been applied in a wide variety of disorders. In this study, the potential effects of adipose-derived MSC (AD-MSCs) therapy and its combination with glucantime were evaluated in a murine model of cutaneous leishmaniasis induced by L. major. The results showed that AD-MSCs improved wound healing and decreased parasite burden. The real-time PCR results obtained from mice treated with AD-MSCs showed that IL-12 and TNF-α genes were upregulated. IL-10, arginase, and FOXP3 genes were downregulated whereas no differences in expression of the IL-4 gene were found. Overall, it seems that AD-MSCs therapy enhances Th1 immune response in L. major infected BALB/c mice. Unexpectedly, our results showed that the association of glucantime to AD-MSCs treatments did not lead to an increment in the anti-leishmanial activity.

First molecular survey of Anaplasma phagocytophilum in hard ticks (Ixodidae) from Southwestern Iran.

Anaplasma phagocytophilum is a gram-negative obligate intracellular tick-borne rickettsia with veterinary and public health importance worldwide. This organism is an etiologic agent of tick-borne fever (TBF) in domesticated animals and human granulocytic anaplasmosis (HGA) as well. Hard ticks (Ixodida: Ixodidae) are incriminated as the main biologic vectors for Anaplasma spp. Studies represent that Ixodes spp. are the main vectors for A. phago-cytophilum and few reports hinted that other tick species may play this role. So, the goal of the presented work was to investigate the A. phagocytophilum in 2000 hard ticks in Khuzestan province of Iran by specific nested-PCR performing two consecutive amplifications of 16SrRNA gene fragment with highly variable nucleotide region. Each reaction included 10 salivary glands of distinct tick species. Specific nested-PCR on accumulated salivary glands detected specific bands in 15.5% of reactions (31 of 200) in electrophoresis only in Rhipicephalus sanguineous and Hyalomma marginatum ticks. We concluded that the broad distribution of A. phagocytophilum infection is not only is due to the existence of Ixodes spp. but other hard ticks may also play a role in this issue.

Anti-leishmanial activity and identification of chemical constituents of Scrophularia striata essential oil by head space solid phase micro extraction gas chromatography mass spectrometry (HS-SPME GC-MS) / Actividad anti-leishmanial e identificación de componentes químicos del aceite esencial de Scrophularia striata mediante microextracción en fase sólida en el espacio de cabeza seguido de cromatografía gaseosa acoplada a espectrometría de masas (HS-SPME GC-MS)

The aim of the present study was to determine the main constituents of Scrophularia striata essential oil and to evaluate in vitro effect of essential oil on Leishmania tropica and Leishmania major promastigotes and axenic amastigotes. Chemical constituents of the extracted essential oil were separated by headspace solid-phase microextraction (HS-SPME) equipped with a PDMS/DVB fiber. The fiber was injected to gas chromatogram- mass spectroscopy (GC-MS) to determine their identity. Finally, after exposure of parasites to different concentrations of water soluble fraction of essential oil, viability of promastigotes and axenic amastigotes were investigated. Based on the HS-SPME results, 47 compounds representing 95.6% of the total oil, were identified in essential oil. Essential oil analysis showed that nonane (19.7%), α-terpineol (17.4%) and linalool (10.2%) were the most abundant compounds. This study indicates that water soluble fraction of S. striata essential oil has promising anti-leishmanial activity.

El objetivo del presente estudio fue determinar los principales componentes del aceite esencial de Scrophularia striata y evaluar el efecto in vitro del aceite esencial en promastigotes y amastigotes axénicos de Leishmania tropica y Leishmania major. Los componentes químicos del aceite esencial extraído se separaron mediante microextracción de fase sólida en el espacio superior (HS-SPME) equipado con una fibra PDMS/DVB. Para determinar su identidad la fibra se inyectó en un cromatógrafo de gases acoplado un espectrómetro de masas (GC-MS). Finalmente, después de la exposición de los parásitos a diferentes concentraciones de fracción soluble del aceite esencial en agua, se investigó la viabilidad de los promastigotes y los amastigotes axénicos. En base a los resultados de HS-SPME, se identificaron 47 compuestos que representan el 95.6% del aceite total en el aceite esencial. El análisis de aceites esenciales mostró que el nonano (19.7%), el α-terpineol (17.4%) y el linalol (10.2%) fueron los compuestos más abundantes. Este estudio indica que la fracción soluble en agua del aceite esencial de S. striata tiene una actividad antileishmanial prometedora.

First Molecular Survey on Anaplasma phagocytophilum Revealed High Prevalence in Rural Dogs from Khuzestan Province, Iran.

BACKGROUND: Anaplasmosis due to Anaplasma phagocytophilum is an important tick-borne zoonotic disease, which affects dogs, horses, cattle and human as well. This study aimed to probe the existence of this organism by means of molecular biology techniques for the first time in rural dogs of Khuzestan province, Southwestern Iran. METHODS: During Sep 2014 to Apr 2015 blood samples of 103 apparently healthy rural dogs (60 males) were collected for A. phagocytophilum detection by light microscopical examination of Giemsa stained slides and Nested PCR on a fragment of 16S rRNA gene. RESULTS: From the examined slides, 11.65% were positive for A. morulae while 57.28% of infection was revealed by Nested PCR method. There was no statistical difference between ages and sexes of dogs and infection in molecular survey of A. phagocytophilum. CONCLUSION: Molecular prevalence of A. phagocytophilum was noticeably high. It may cause the incidence of disease in human population.

Sperm quality and hormonal levels in C57BL/6 mice infected with Neospora caninum.

Neospora caninum is a major pathogen of cattle and dogs, and neosporosis is widespread in five continents. In this study effect of experimental neosporosis on sperm quality of C57BL/6 mice in different days was investigated. Based on the results sperm concentration was not changed in infected groups but neosporosis induced a significant decrease in epididymis sperm motility at 60 days post infection. A significant increase in the number of abnormal sperms at five, 15, 30 and 60 days post infection was found. At days 15, 30 and 60 post infection testosterone concentrations were significantly low in infected groups and FSH level was significantly high in infected groups at five and 30 days post infection. LH level was decreased in infected groups, but the difference was significant at five, 15 and 30 days post infection. Comparison of TSH and T4 levels between groups revealed a significant decrease in infected groups at five, 15, 30 and 60 days post infection. Except 15 days post infection T3 levels decreased significantly in infected groups. GPX activity, MDA and TAC level was significantly increased in infected mice at five days post infection. In this study neosporosis is associated with hypogonadotrophic gonadal insufficiency in infected C57BL/6 male mice.