Molecular identification of Cryptosporidium, Giardia, and Blastocystis from stray and household cats and cat owners in Tehran, Iran.

Cryptosporidiosis, giardiasis, and blastocystosis are among the most important parasitic diseases common between humans and cats. In addition, there are concerns about the possible transmission of zoonotic parasites from infected cats to humans. Hence, we investigated the molecular epidemiology of Cryptosporidium spp., Giardia duodenalis, and Blastocystis sp. in stray and household cats and cat owners. Our study was performed on 132, 33, and 33 fecal samples of stray and household cats, as well as cat owners in Tehran, Iran. Cryptosporidium spp. was identified using a nested PCR targeting the small subunit ribosomal RNA gene (SSU rRNA) and sequencing the internal amplified fragments. Furthermore, to perform multilocus genotyping of G. duodenalis, the ß-giardin (bg), glutamate dehydrogenase (gdh), and triosephosphate isomerase (tpi) genes were amplified to assess the DNA of G. duodenalis in the fecal samples of cats and cat owners. In addition, Blastocystis was detected by targeting the SSU rRNA gene, and the subtypes of Blastocystis were determined via the sequencing of amplicons. Cryptosporidium felis and Cryptosporidium canis were detected in seven stray cats (5.3%) and one household cat (3%). The bg gene of G. duodenalis was amplified and successfully sequenced in two (1.5%) stray cats and revealed assemblages F and B of G. duodenalis. Sequencing and phylogenic analysis of SSU rRNA gene nucleotide sequences of Blastocystis detected ST5 and ST10 in stray cats (1.5%), ST1 in household cats (9.1%), and ST1, ST2, ST3, and ST7 in owners (30.3%). The low prevalence of Cryptosporidium, Giardia and Blastocystis in cats and the presence of species/assemblages/subtypes with limited zoonotic potential indicate that cats had a minor role in their owners' infection in the investigated population. However, the presence of zoonotic protozoa in cats suggests the necessity of special attention to high-risk individuals during close contact with cats. Therefore, it is recommended that veterinarians, physicians, and urban managers plan to prevent, control, or treat these parasites to help the urban community live healthily alongside cats.

Effects of negative air ions (NAIs) on Leishmania major: A novel tool for treatment of zoonotic cutaneous leishmaniasis (ZCL).

BACKGROUND: Cutaneous leishmaniasis (CL) is a Neglected Tropical Disease (NTD) that causes high morbidity in the tropics and sub-tropics. Despite the remarkable advancements in the treatment of CL, the available therapeutics are far from ideal and also cause serious adverse side effects. Negative air ions (NAIs) generators are widely available for domestic and industrial uses. Several studies have reported on positive effects of NAIs therapy on human health as a non-pharmaceutical treatment for respiratory disease, allergy, or stress-related health conditions, including infectious diseases. To our knowledge, no studies have examined the effectiveness of the NAIs therapy against Leishmania parasites. The aims of this study were to investigate the effect of NAIs therapy on Leishmania major (L. major) the causative agent of CL in in vitro and in a murine model. METHODOLOGY/PRINCIPAL FINDINGS: In vitro anti-leishmanial effects of NAIs therapy were measured by parasitological methods. NAIs therapy was assessed in vivo in L. major infected BALB/c mice by measuring the footpad (FP) lesion size and parasite load using metric caliper tool and qPCR, respectively. Immune responses in treated and non-treated mice were assessed by measuring the levels of IFN-γ, IL-4, NO and arginase activity. In vitro NAIs therapy significantly decreased the viability of Leishmania promastigotes and of amastigotes cultured in macrophages, but did not affect the host cells. NAIs therapy of L. major infected BALB/c mice resulted in reduced FP lesion size, diminished parasite burden, and importantly decreased induction of IL-4 and arginase activity in the presence of NAIs. In contrast IFN-γ and NO levels were significantly enhanced. NAIs therapy significantly diminished the progression of disease compared to the control group, but was less effective than amphotericin B treatment. CONCLUSIONS: Our study shows that NAIs treatment was effective in vitro and in Leishmania-infected mice, elicited a T-helper 1 (Th1) response and increased efficient cellular immunity, resulting in a diminished parasite load. Therefore, NAIs therapy can be considered as a useful and safe tool that can contribute to clearing L. major infections without inducing toxicity in host cells. The applications and mechanisms of NAIs therapy warrant further investigation especially in humans suffering from CL.

Prevalence and Molecular Characterization of Toxoplasma gondii and Toxocara cati Among Stray and Household Cats and Cat Owners in Tehran, Iran.

Toxoplasma gondii and Toxocara spp. are the most critical parasites common between humans and cats. The close association of cats with humans in urban areas persuaded us to investigate the prevalence of these parasites in stray and household cats and their possible role in the owners' infection. Herein, 132 and 33 fecal samples of stray and household cats, respectively, and 33 blood samples of their owners were collected in Tehran, Iran. The prevalence of T. gondii was determined by targeting the B1 gene in the feces of stray and household cats and the blood of cat owners. Furthermore, genotypes of T. gondii were identified based on the multilocus genotyping of BTUB, GRA6, SAG3, and APICO loci. Toxocara spp. were detected by targeting the second internal transcribed spacer (ITS-2) of the ribosomal DNA of these parasites in the cats' feces and the humans' blood. Also, Toxocara IgG was assessed in the human serum samples. The B1 gene amplification showed that 15.2% of stray cats, 18.2% of household cats, and 51.5% of cat owners were infected with T. gondii. The multilocus sequence analysis revealed the predominance of genotype I of T. gondii in stray cats and genotype II of T. gondii in household cats and cat owners. The amplifying of ITS-2 revealed a high prevalence of T. cati infection (47.0%) in stray cats, whereas no infection was found in the feces of household cats or the serum of cat owners. Likewise, Toxocara IgG was not detected in the serum of humans. The lower prevalence of T. gondii in stray/household cats than in the cat owners indicates the limited impact of close contact with infected cats in human toxoplasmosis. However, the high prevalence of T. cati infection in stray cats can cause contamination of the environment by excreting eggs that may lead to infecting humans through soil or water. Therefore, public health education in urban management planning is necessary for routine urban cat deworming programs and for training the healthcare workers to prevent, control, and treat these infections.

Molecular Identification and Genotyping of Babesia canis in Dogs from Meshkin Shahr County, Northwestern Iran.

BACKGROUND: Canine babesiosis is one of the mainly worldwide-distributed tick-borne haemoprotozoan parasitic diseases in dogs. METHODS: A total of 43 blood samples were randomly collected from naturally infected dogs in seven villages from different geographical areas of Meshkin Shahr, Ardabil Province, Iran. The presence of Babesia species detected with standard methods including parasitological and gene sequencing techniques targeting the 18S rRNA gene. RESULTS: Our results revealed that four dogs 9.3% (4/43) including one female and three male dogs were infected with Babesia. All four Babesia-infected dogs were confirmed B. canis by the molecular-based method. Sequence alignments comparison of the B. canis genotypes A and B, it was revealed that all B. canis isolates belonged to genotype B. CONCLUSION: This study provides essential data for subsequently define the critical importance of the molecular studies in management and prevention of the canine babesiosis in Iran.

Effect of Otostegia persica extraction on renal injury induced by hindlimb ischemia-reperfusion: a rat model.

INTRODUCTION: It is known that ischemia-reperfusion causes remote organ injury as well as local injury. In traditional systems of medicine, many plants have been documented to be useful for the treatment of various disorders including oxidative esters. This study was designed to investigate whether Otostegia persica extraction pretreatment has a protective effect against renal injury induced by hindlimb ischemia-reperfusion. METHODS: Forty male Wistar rats were allocated into five groups as follows: Control, Sham, Otostegia persica, ischemia-reperfusion and ischemia-reperfusion+Otostegia persica groups. Rats in Otostegia persica and ischemia-reperfusion+Otostegia persica groups received Otostegia persica extraction (300 mg/kg) orally 2 days prior to operation. Hindlimb ischemia was induced by clamping the femoral artery for 2 h. After 24 h of reperfusion, blood and urine samples were obtained for kidney function tests and the kidneys were removed for histological analysis and oxidative stress measurement. RESULTS: The decrease in glomerular filtration rate induced by reperfusion was significantly improved by Otostegia persica extraction administration (P<0.05), which resulted in the decrease in serum urea and creatinine concentrations. Urinary creatinine significantly decreased in ischemia-reperfusion group compared to the other groups (P<0.05). Urinary excretion rate, water intake and the ratio of kidney/body weight significantly increased in animals with reperfusion injury as compared with other groups (P<0.05). On histological examination, rats pretreated with Otostegia persica extraction had nearly normal morphology. Skeletal muscle ischemia-reperfusion produced a significant increase in renal tissue malondialdehyde level, while pretreatment with Otostegia persica extraction was associated with a significantly lower malondialdehyde level (P<0.05). Renal tissue catalase and superoxide dismutase activity and glutathione level were significantly (P < 0.05) decreased by hindlimb ischemia-reperfusion. The increases in these parameters were decreased by pretreatment with Otostegia persica extraction. CONCLUSIONS: The results of this study showed that Otostegia persica extraction pretreatment significantly protected the renal injury from skeletal muscle ischemia-reperfusion.

Influence of tramadol on ischemia-reperfusion injury of rats' skeletal muscle.

INTRODUCTION: Tramadol has been shown to decrease ischemia-reperfusion injuries in myocardial or brain tissues. The aim of this study was to assess the effects of tramadol on ischemia-reperfusion injury in a rat hind limb ischemia-reperfusion model. METHODS: Forty-five healthy adult male Wistar rats were randomized into three experimental groups as follows: Sham, Ischemia-reperfusion and Ischemia-reperfusion + tramadol groups. Ischemia was induced in anesthetized rats by left femoral artery clipping for 2 h followed by 24 h of reperfusion. Tramadol (20 mg/kg) was administered intravenously immediately prior to reperfusion. Blood pH, pO2, pCO2, HCO3, creatine phosphokinase (CPK), lactate dehydrogenase (LDH) as well as plasma malondialdehyde (MDA) were measured at the end of the reperfusion. Left gastrocnemius muscle samples were taken for histological and biochemical examination. RESULTS: The pH and pCO2 were similar in all study groups, with no statistical significance. pO2 and HCO3 levels presented the highest elevation in sham and Ischemia-reperfusion + tramadol groups, as compared to Ischemia-reperfusion group (P < 0.05). The extent of muscle changes in the ischemia-reperfusion + tramadol group was significantly lower than ischemia-reperfusion group (P < 0.05). In comparison with other groups, serum and tissue MDA levels in ischemia-reperfusion group were significantly increased (P < 0.05). The muscle tissue glutathione (GSH), superoxide dismutases (SOD) and catalase (CAT) levels in the Ischemia-reperfusion group were significantly lower than the other groups (P < 0.05). Wet/dried weight ratio in ischemia-reperfusion group was significantly higher (P < 0.05) than subjects in other groups. CONCLUSIONS: From the histological, histochemical and serum biochemical perspective, the treatment with tramadol has alleviated the metabolic injuries in the skeletal muscle ischemia and reperfusion in this experimental model.