Cellulitis Caused by Hirudo orientalis Bites That Lead to an Allergic reaction.

The allergic reaction due to leech bites is frequently reported due to complications of leech therapy and also unwanted leech infestation. Regularly, the urticarial papules are common, and itching lasts less than 24 h. In the case of leech biting, dermal infection could be caused by leech gut bacterial flora such as Aeromonas spp and histamine from leech saliva. In this case report, a 30-year-old diabetic woman, who works in the field of leech breeding, was bitten by Hirudo orientalis during breeding. Her clinical signs were inflammation, swelling, pain, and redness in the back of her left hand. A microbiological examination revealed that the isolated leech was infected with Aeromona hydrophila. The risk of death due to anaphylactic shock and sepsis is high in some cases of underlying diabetes and immunocompromised individuals. The study pointed out the hazards of leech bites and proposed preventative measures such as using gloves and boots for farm workers.

Morphological and Molecular Descriptions of Macracanthorhynchus ingens (Acanthocephala: Oligacanthorhynchidae) Collected from Hedgehogs in Iran.

Aim: Limited data exist on acanthocephalan infections of hedgehogs in the world. Our objective was to investigate the prevalence and distribution of Macracanthorhynchus ingens infection in hedgehogs between August 2021 and March 2022 (n = 30) in the east of Iran. Methods: At first, infection with M. ingens was diagnosed based on morphologic features of the adults such as body length, proboscis, and hooks. Spindle-shaped eggs (mean length, 99.1 microns; mean width, 60.1 microns) were obtained from the body cavity of gravid female specimens. Results: The molecular analysis based on 18S rDNA and COX 1 genes confirmed the morphological identification of isolated M. ingens. The prevalence of M. ingens in our sample was 13.3% with 1-10 worms per infected host. Conclusion: In this study, we identify M. ingens as zoonotic species in hedgehog carcasses for the first time that passed eggs and adult worms, indicating parasite maturation and reproduction. There are a few studies on acanthocephalans in Iran. Therefore, more comparative studies are needed to determine the status of these species.

Nano amphotericin B: a good anti-leishmanial drug with effect on cathelicidin gene expression.

Protozoan parasites, such as Leishmania major (L. major), remained as a global health problem of the current century. Leishmania major is a major cause of cutaneous leishmaniasis (CL) in developed and developing countries. Traditionally, amphotericin B is prescribed as an alternative drug, while first-line drugs failed. Some active proteins of the innate immune system such as cathelicidins try to inhibit infection Via several proposed mechanisms. Here this research aimed to not only determine the anti-leishmanial activity of nano amphotericin B but also to evaluate which anti-leishmanial drug can induce the cathelicidin gene expression. Both promastigote and amastigote stages of L. major were exposed to various concentrations of nano amphotericin B, amphotericin B and finally compared to glucan time as standard drug for CL treatment. For the gene expression of cathelicidin, macrophages were exposed to the same concentration of anti-leishmanial drugs. The findings demonstrated that nano amphotericin B was more effective at all concentrations than amphotericin B. Additionally, among tested anti-leishmanial drugs, nano amphotericin B has more potency to induce the cathelicidin gene expression in macrophages cells. The findings revealed that nano amphotericin B has potential as an effective anti-leishmanial drug against CL caused by L. major parasites.

Frequency of different genotypes of Giardia duodenalis in slaughtered sheep and goat in east of iran.

Giardia duodenalis is one of the most common and important protozoan parasites of the gastrointestinal tract in humans and animals, especially in developing countries. The purpose of this study was determining prevalence of Giardia genotypes specially zoonosis genotypes in sheep and goat in eastern of iran slaughterers.This cross-sectional study was conducted during April to November 2019. 300 fecal samples were collected from the rectum of sheep and goats. The samples were subjected to DNA extraction after sucrose gradient purification. A fragment of the glutamate dehydrogenase gene (gdh) was amplified by semi-nested PCR and genotype diagnosis was performed by digestion of the secondary PCR product with restriction enzymes RsaI and Nla IV. The prevalence of Giardia was found as (274/300) by the molecular method. Restriction endonuclease digestion of the nested-PCR product showed; among 274 positive isolates, 95 were typed as assemblage E, 15 as assemblage B, 87 assemblage AI, 45 assemblage AII, and 32 assemblege C. In this study, frequency of different assemblages of G. duodenalis was determined in sheep and goats by gdh gene and PCR-RFLP method. Same of other studies, assemblage E was dominant genotype in sheep and goats. Isolation of zoonotic assemblages as AI, AII, and BIII showed that sheep and goats should be considered as a source for human infection.

Murine cathelicidin: as a host defensive response against Leishmania major infection.

Leishmaniasis is a serious global challenge with neither efficacious prophylactic vaccine nor effective and safe therapeutic measures. Cathelicidins, members of antimicrobial peptides family, are small proteins of innate immunity system, which represent a protective barrier against a number of potential pathogens in living organisms. The murine cathelicidin or cathelin-related antimicrobial peptide (CRAMP) is expressed by a variety of cells or tissues, and highly resembles to human cathelicidin (LL-37). It is naturally expressed at a low concentration in adolescent age, but extensively increases during cutaneous infections. Despite its important role, it has less been investigated in parasitic infections. Among all cells, macrophages and skin cells are the two important cells that directly have a relationship with Leishmania major parasites. The present study aimed to show whether cathelicidins protect their hosts following cutaneous leishmaniasis due to L. major parasites. Both in vitro and in vivo models of L. major infection were established by exposing of J744 cell line (murine macrophages) and BALB/c mice with the stationary phase of L. major promastigotes for 24 h and 7 days. The findings revealed that both macrophages and skin cells significantly (p < 0.05) expressed a high level of CRAMP gene and peptide after challenging with L. major parasites. Thus, our data suggest a protective role for cathelicidins against infections caused by L. major parasites. This experimental model could be considered as a novel potential vaccine candidate for planning future control strategy against human leishmaniasis.

Efficacy and Safety of Zataria multiflora Boiss Essential Oil against Acute Toxoplasmosis in Mice.

BACKGROUND: Broad spectrums of pharmacological properties, including antimicrobial activity have been attributed to Zataria multiflora Boiss (Laminaceae). The in vivo efficacy and safety of Z. multiflora essential oil (ZM-EO) were evaluated against acute toxoplasmosis caused by Toxoplasma gondii (Sarcocystidae) in mice. METHODS: Z. multiflora (aerial parts) was obtained from the rural districts of Kerman city (Kerman Province) Southwestern Iran, in May of 2016. Male NMRI mice were orally treated with normal saline (control group) and ZM-EO at the doses of 0.2 and 0.4 mL/kg once a day for 14 d (8 mice in each group). On the 15th day, the mice were infected with 104 tachyzoites of T. gondii RH strain by intraperitoneal route. The mortality rate and parasite load were determined in the infected mice. Additionally, 24 mice were applied to examine the sub-acute toxicity of ZM-EO at the above doses after treatment during 14 d. RESULTS: GC/MS analysis displayed that the key constituents were thymol (45.4%), carvacrol (23%) and p-cymene (10.6%), respectively. Overall, 100% mortality was observed on the 8th and 9th days in treated mice with the concentrations of 0.2 and 0.4 mL/kg, respectively. The mean number of tachyzoites in the mice treated with 0.2 and 0.4 mL/kg of ZM-EO were 189×104 and 76×104 cell/mL, respectively, meaningfully (P<0.05) reduced compared with the control mice. Results also demonstrated that ZM-EO had no important toxicity on mice. CONCLUSION: The results demonstrated the efficacy of ZM-EO against acute toxoplasmosis. Nevertheless, supplementary surveys are mandatory to examine its precise effects, mainly immunomodulatory effect on toxoplasmosis.

Efficacy and safety of Curcuma longa essential oil to inactivate hydatid cyst protoscoleces.

BACKGROUND: One of the most important ways to cure hydatid cysts is surgery. Today, available chemical drugs have shown severe complications during hydatid cyst surgery. Here we investigated theefficacy and safety of Curcuma longa (C. longa) essential oil (CLEO) against hydatid cysts protoscoleces. METHODS: Here, the collected protoscoleces from fertile hydatid cysts were exposed to different concentrations of the CLEO (50-200 µl/mL) for 5-30 min in vitro and ex vivo. In addition, male NIH mice (48 mice) were applied to examine the safety of CLEO. RESULTS: All protoscoleces were completely killed in 5th min of treatment to the doses of 200 µl /mL of CLEO. On the other hand, in the 10th min of treatment, CLEO entirely killed 100% of protoscoleces at the dose of 100 µL/mL. Other doses of CLEO, but, revealed a postponed activity. Although CLEO at the doses of 50, and 100 µL/mL exhibited no similar effect in the ex vivo analysis; but, at the dose of 200 µL/mL and an exposure time of 5 min, approximately 100% of protoscoleces were destroyed into the cyst. We found that after intra-peritoneal injection of the CLEO for 14 days, although blood and biochemical parameters were changed; but there was no significant difference in comparison with the control group (p > 0.05). CONCLUSION: This research demonstrated that C. longa revealed the promising scolicidal effects against protoscoleces in vitro and in vivo, of course, after additional tests; it might be considered as an herbal scolicidal drug in order to decrease the threat of discharge of protoscoleces through hydatid cyst surgery. Nevertheless, supplementary studies will be desired to prove the current findings by examination the essential oil in a clinical setting.

Thermotolerant Acanthamoeba spp. isolated from recreational water in Gorgan City, north of Iran.

Acanthamoeba as free-living parasites are scattered ubiquitously, throughout the world. This study was aimed to evaluate the presence of Acanthamoeba spp. genotypes in the recreational water sources in Gorgan County, the capital of Golestan Province using both morphological and molecular approaches. Thirty water samples were collected from different recreational waters in Gorgan, the capital of Golestan Province, northern Iran during 2015-2016. Samples were filtered and followed by culture in non-nutrient agar. Acanthamoeba were identified both by morphological and molecular analysis. The pathogenical potential of positive cloned samples were also determined using tolerance test. Twenty-six percent of recreational water were identified as Acanthamoeba spp. based on the morphological analysis and from these positive samples, five samples were successfully sequenced after molecular studies. Phylogenetic analysis showed the clustering of four samples in T4 genotype group and only one sample as T15 genotype. Thermotolerance test revealed that all cloned samples were highly positive. Since the attractiveness of recreational places for people is increasing, the potential risk of this water should be monitored routinely in each region. More studies are needed to better evaluate the risk of this ubiquitous parasite for the human.

The First Survey of Isolation and Molecular Typing of Toxoplasma gondii by Bioassay and PCR Method in BALB/c Mice in Camels (Camelus dromedarius) from Eastern Iran.

BACKGROUND: We isolated Toxoplasma gondii from camels by bioassay method in mice model and detect parasitic DNA in brain mice by molecular methods. METHODS: One hundred tissue samples including heart (n=50), and diaphragm (n=50) were collected from camels (n=50) slaughtered in abattoirs from Feb to Oct 2015 in three provinces located in eastern Iran. In first, blood sample from 50 camels was assayed for anti-Toxoplasma antibodies by modified agglutination test (MAT) test. Bioassay method was done in positive MAT blood camels in BALB/c mice and Nested PCR performed in seropositive tissue samples to amplify the B1 and GRA6 genes. The existence of polymorphic restriction sites for endonuclease MseI was used with PCRRFLP method and Sequencing analysis to evaluate the prevalence of type strains (I, II and III). RESULTS: Overall, 13 (26%) of camels were positive with titer of 1:20 for toxoplasmosis and 13(26%) tissue samples of camels were found positive for the T. gondii B1 gene, including 7(14%) diaphragm, 6(12%) heart. Moreover, 3(6%) tissue samples of camels were found positive with GRA6 gene for T. gondii. There are three genotypes and mix genotype using MseI enzyme among all positive samples. CONCLUSION: The obtained results from serological and molecular tests demonstrated the infection of T. gondii with previously recognized genotypes in the tissues of camels for first time from Iran. Since consumption of meat camels are raising in Iran, there may be a high risk of toxoplasmosis through consumption of products from these hosts due to their susceptibility to the infection.

Prevalence and Risk Factors of Pediculosis in Primary School Children in South West of Iran.

BACKGROUND: Pediculosis or louse infestation is a public health problem in many developing countries where the WHO's primary health-care program is inefficient and haphazard. The present study aimed to determine the prevalence of Pediculus capitis infestation and its related risk factors in the primary school children in Andimeshk, Dezful and Shoosh counties; Khuzestan Province, Iran. METHODS: Overall, 28410 students in the age range of 7-11 years old in primary schools of North of Khuzestan Province, southern Iran were examined individually and privately under the flash light for all life cycle stages of lice or their nits in 2016. A questionnaire was filled for each school child before hair examination; then examination was carried out to detect head lice as well as eggs/nits. RESULTS: Overall, 2995 students (10.5%) were infested with pediculosis. There was a significant difference in the prevalence of pediculosis among the boys and girls students. The prevalence of infestation was also significantly (P<0.05) higher in students of living in nomad tribes (23.8%) in comparison of rural (12.4%) and urban areas (6.5%). CONCLUSION: Several risk factors significantly (P<0.05) related to pediculosis included gender of female, nomad habitat, parents education, father's occupation, having health staff, history of infestation and number of combing per day. Increasing awareness and training of teachers and relatives, as well as for improving standards of personal health, can significantly reduce the prevalence of pediculosis.

Host-parasite Responses Outcome Regulate the Expression of Antimicrobial Peptide Genes in the Skin of BALB/c and C57BL/6 Murine Strains Following Leishmania major MRHO/IR/75/ER Infection.

BACKGROUND: Different background of immunity responses determine resistance or susceptibility of certain mouse strains to Leishmania major infection. Some have been well known previously. Antimicrobial peptides (AMPs) such as cathelicidins and defensins are unique fragments of innate immunity system with well-known effects against the invasion pathogens. Despite their outstanding roles and being of extensive cases of cutaneous leishmaniasis (CL) caused by L. major, they have been less studied in Leishmania fields. The aim of present study was to determine whether these components play a role in the protection of skin against Leishmania infections. METHODS: The animal model of Leishmania infection was established by the subcutaneous inoculation of 5×106(parasites/ml) from the stationary phase of L. major promastigotes to BALB/c and C57BL/6 mice from January 2016 to August 2016 in Kerman Province, southeast of Iran. After 1, 3 and 7 d of post-infection (PI), the samples needed for detecting of the mRNA levels of mouse beta-defensin (mBD)-1, mBD2, mBD3, mBD4, mBD6, cathlin-related antimicrobial peptide (CRAMP), interleukin (IL)-10, IL-12 and parasite load were taken under standard methods. RESULTS: The findings related to cytokines profiles in BALB/c (↑IL-10, ↓IL-12) and C57BL/6 mouse strains (↓IL-10, ↑IL-12) demonstrated that immunity system has been accurately activated during CL caused by L. Major parasites. We also observed a significant up-regulation of all aforementioned AMPs genes in BALB/c mice at selected times compared to another strain. CONCLUSION: CL occurred in BALB/c mice in spite of the fact that the expression of AMPs was higher than the other strain. AMPs genes are well expressed to provide defense against the parasites that have increased and escaped from immunity system but cannot create an absolute protection.

Considerable Genetic Diversity of Trichomonas vaginalis Clinical Isolates in a Targeted Population in South of Iran.

BACKGROUND: The present study aimed to characterize genetically and to compare the most frequently occurring strains of Trichomonas vaginalis isolated from southern Iran. METHODS: Totally, 150 vaginal swab and urine specimens were collected from symptomatic and asymptomatic women from May 2012 to Jun 2013. This study implemented a sensitive and reliable PCR-restriction fragment length polymorphism (RFLP) typing method on the actin gene. Moreover, one representative sample of each identified genotype was subjected to sequencing. RESULTS: Twenty-four T. vaginalis isolates were positive and 6 distinct electrophoretic patterns (H, E, G, I, M, N) were identified. Genotypes H and I were found to be more prevalent (50 and 37.5%) in Kerman and Shiraz, respectively. The phylogenetic analysis showed that two isolates were located as a separated clade with the other T. vaginalis isolates. CONCLUSION: The obtained findings showed a considerable genetic polymorphism of clinical isolates from the population studied. More studies may be warranted in future as to unveiling any possible links between a given genotype/cluster and pathogenic behavior of T. vaginalis.

Prevalence of Toxoplasma gondii Infection among Healthy Blood Donors in Northeast of Iran.

BACKGROUND: This cross-sectional investigation aimed to evaluate the prevalence of IgM and IgG anti-Toxoplasma gondii antibodies and the associated risk factors among healthy blood donors in Khorasan Razavi Province, northeast of Iran from Nov 2014 to May 2015. METHODS: Overall, 491 serum samples from apparently healthy blood donors referred the six biggest blood centers in Razavi Khorasan, Iran, were screened for IgG and IgM anti-T. gondii antibodies by enzyme-linked immunosorbent assay (ELISA). A structured questionnaire was used to obtain information on risk factors for T. gondii infection. Nested PCR was also used to detect DNA of T. gondii in the IgM-positive samples by using of B1 and RE (Repetitive Element) as marker for amplifying fragment size of 531 bp and 164 bp in PCR method. RESULTS: Totally, 200 (40.7%) samples were seropositive for anti-T. gondii antibodies; 184 (37.5%) donors tested seropositive for only IgG antibody, 8 (1.6%) tested seropositive for both IgM and IgG and 8 (1.6%) were positive for IgM antibody alone. Several risk factors significantly related to T. gondii seropositivity in the univariate analysis at P<0.05 included age (P<0.001), and raw/half-cocked meat consumption (P=0.015). T. gondii DNA was found in all sixteen IgM-positive samples. CONCLUSION: T. gondii infection was present among healthy blood donors in northeast of Iran. Thus, it is suggested to design screening programs for preventing transfusion-transmitted toxoplasmosis.

Zataria multiflora Bioss: lethal effects of methanolic extract against protoscoleces of Echinococcus granulosus.

The present study was aimed to evaluate the lethal effects of Zataria multiflora Boiss (Lamiaceae) methanolic extract against Echinococcus granulosus protoscoleces. Protoscoleces were aseptically aspirated from sheep livers having hydatid cysts. Various concentrations of the essential oil (2.5-20 mg/mL) were used for 10-60 min. Viability of protoscoleces was confirmed using eosin exclusion test (0.1 % eosin staining). Obtained results showed that Z. multiflora extract at the concentration of 20 mg/mL after 10 min of exposure killed 100 % protoscoleces. The mean of mortality rate of protoscoleces after 20 min of exposure to the concentration of 10 mg/mL was also 100 %. Lower concentrations of Z. multiflora extract provoked a delayed protoscolicidal activity. The findings indicated potential of Z. multiflora methanolic extract as a natural source for the producing of new scolicidal agent for use in hydatid cyst surgery.

Seroprevalence of Toxoplasma gondii infection among childbearing age women in Kerman city, southeastern Iran.

This cross-sectional study aims to determine the prevalence of IgM and IgG anti-T. gondii antibodies and the associated risk factors among childbearing age women referring to counseling centers before marriage in Kerman city, southeast of Iran. Totally, 300 serum samples were collected from women referred to Central Laboratory for Marriage Consultation in Kerman city were screened for IgG and IgM anti-T. gondii antibodies by enzyme linked immunosorbent assay (ELISA). Out of the 300 serum samples, 38 (12.6 %) tested seropositive for anti-T. gondii antibodies; 31 (10.3 %) samples tested seropositive for only IgG antibody, 1 (0.33 %) tested seropositive for both IgM and IgG and 6 (2.0 %) were positive for IgM antibody alone. Statistical analyses also indicated that seroprevalence of anti-T. gondii antibodies increased with age (p < 0.05). Moreover, some risk factors such as, living in rural regions, contact with cats, raw/half-cooked meat consumption, and agricultural activities were significantly (p < 0.05) related to T. gondii seropositivity. The findings revealed that more than three-quarters of the childbearing age women studied in the present investigation are susceptible to infection during pregnancy. Thus, by adopting correct and improved practices we can improve their living conditions, and prevent infection and awareness and control of pathogens associated with disease is recommended.

Efficacy and Safety of Bunium Persicum (Boiss) to Inactivate Protoscoleces during Hydatid Cyst Operations.

BACKGROUND: Current scolicidal agents, which have been used for inactivation of protoscoleces during surgical procedures, are associated with adverse side effects including sclerosing cholangitis. This investigation aimed to evaluate the scolicidal effects of Bunium persicum (Boiss) essential oil against protoscoleces of hydatid cysts and also its toxicity in a mice model. METHODS: Protoscoleces were aseptically aspirated from sheep livers having hydatid cysts. Various concentrations of the essential oil (3.125-50 mcL/mL) were used for 5-30 min. The viability of protoscoleces was confirmed using the eosin exclusion test (0.1% eosin staining). Further, 48 male NMRI mice were used to determine the acute and sub-acute toxicity of B. persicum essential oil. RESULTS: The obtained results revealed that the B. persicum essential oil at the concentrations of 25 and 50 mcL/mL after 5 min of exposure killed 100% protoscoleces. The mean mortality rate of protoscoleces after 10 min of exposure to the concentration of 12.5 mcL/mL was 100%. Lower concentrations (6.25 and 3.125 mcL/mL) of B. persicum essential oil, however, indicated a delayed protoscolicidal effects. The LD50 value of intra-peritoneal injection of the B. persicum essential oil was 1.96 mL/kg body wt. No significant difference (p > 0.05) was observed in the clinical chemistry and hematologic parameters after oral administrations of B. persicum essential oil at the doses 0.05, 0.1, 0.2, and 4 mL/kg for 14 d. CONCLUSION: Our findings demonstrated the potent scolicidal activity of B. persicum with no significant toxicity; it might be used as a natural scolicidal agent in hydatid cyst surgery.

Chemical composition, protoscolicidal effects and acute toxicity of Pistacia atlantica Desf. fruit extract.

The present study was designed to evaluate the chemical composition and scolicidal effects of Pistacia atlantica Desf. extract against protoscoleces of hydatid cysts and its acute toxicity in mice model. Various concentrations of the methanolic extract (5-50 mg/mL) were used for 10-60 min. Viability of protoscoleces was confirmed using eosin exclusion test (0.1%). Acute toxicity was also determined in mice model. The main components were ß-myrcene (41.4%), α-pinene (32.48%) and limonene (4.66%). Findings demonstrated that P. atlantica extract at the concentrations of 25 and 50 mg/mL after 20 and 10 min of exposure killed 100% protoscoleces. The LD50 of the intraperitoneal injection of the P. atlantica methanolic extract was 2.43 g/kg and the maximum non-fatal dose was 1.66 g/kg. Obtained results showed the potential of P. atlantica extract as a natural source with no significant toxicity for the production of new scolicidal agent to use in hydatid cyst surgery.

Efficacy of the Bunium persicum (Boiss) Essential Oil against Acute Toxoplasmosis in Mice Model.

BACKGROUND: We evaluated the in vivo activity of Bunium persicum (Boiss) essential oil on infected mice with acute toxoplasmosis. METHODS: To evaluate prophylactic effects, male NMRI mice received B. persicum essential oil at the concentrations of 0.05 and 0.1 mL/kg for 14 days. After 24 h mice were infected intraperitonealy with 1×10(4) tachyzoites of T. gondii, RH strain. In order to investigate therapeutic effects, mice were infected and then received B. persicum oil at the concentrations of 0.05 and 0.1 ml/kg two times a day for 5 days. The time/mean time of death in all infected mice and the number of tachyzoites from infected mice were recorded. RESULTS: The time/mean time of death of infected mice was 8 and 9 days after oral administration of B. persicum oil at the concentration of 0.05 and 0.1 mL/kg, respectively (P<0.05). In contrast, the time/mean time of death control group was 5 days. In addition, B. persicum significantly reduced the mean number of tachyzoites compared with control group. The time/mean time of death of infected mice was 6 and 7 days after oral administration of B. persicum essential oil at the concentration of 0.05 and 0.1 mL/kg, respectively. In contrast, the time/mean time of death control group was 5 days. B. persicum especially at the concentration of 0.1 ml/kg significantly reduced the mean number of tachyzoites compared with control group. CONCLUSION: The results showed the potential of B. persicum essential oil as a natural source for the production of new prophylactic agent for use in toxoplasmosis.