Inhibitory Effect of Hemiscorpius lepturus Scorpion Venom Fractions on Tachyzoites of Toxoplasma gondii.

Background: The present study determined the effect of the fractions obtained from Hemiscorpius lepturus scorpion venom on the tachyzoite of Toxoplasma gondii. Methods: The fractions of dried venom of He. lepturus scorpion of Khuzestan Province, southern Iran in 2019 were isolated through gel filtration chromatography, and then tachyzoites were exposed to fractions of venom at different concentrations. Trypan blue counting and MTT were applied to assay tachyzoite viability, and the inhibition of the cellular growth of fractions in Vero cells was evaluated. Results: The maximum effect on tachyzoite was observed in fraction 5 of venom. To further separate the protein, fraction 5 was used in high-performance liquid chromatography assay to purify its proteins. Based on the results of HPLC of fraction 5, among which the second peak, a peptide with <10 KDa representing a more potent effect in eliminating the tachyzoite of T. gondii. Conclusion: The scorpion venom-purified fractions possess anti-parasitic activity against the tachyzoite of T. gondii and can be used in parasite-controlling studies.

Nephrotoxicity of Albendazole and Albendazole Loaded Solid Lipid Nanoparticles in Mice with Experimental Hydatidosis.

Purpose: Cystic echinococcosis (CE) is a serious contemporary public health problem. Different CE treatment methods are of considerable importance, with albendazole (ABZ) being one of the most preferred drugs for CE treatment and prophylaxis. In this study, we evaluated the nephrotoxicity caused by ABZ and ABZ-loaded solid lipid nanoparticles (SLNs) in mice with experimental hydatid cyst. Methods: ABZ-loaded SLNs were produced by micro-emulsification and a high shear homogenization technique. Thereafter, we evaluated the physicochemical characterization of the product. Live protoscolices were injected into mice to induce experimental hydatidosis. Mice were then treated with ABZ and ABZ-loaded SLNs. The nephrotoxicity effects were evaluated by biochemical and histopathological surveys. Results: Significantly different blood urea nitrogen (BUN) levels were observed between the two infected groups (ABZ treatment and ABZ-loaded SLN treatment) and the control group. The kidney malondialdehyde (MDA) and glutathione (GSH) levels of the infected groups were not significantly different from those of the control group. The histopathological study revealed nephropathic and pathologic changes in the ABZ and ABZ-loaded SLN groups. Conclusion: ABZ formulated for ABZ-loaded SLNs had a more prominent chemoprophylactic efficacy on CE and fewer side effects than ABZ alone. Neither ABZ nor ABZ-loaded SLNs caused significant biochemical and histopathological defects on the kidney, and all functional biochemical markers stayed within the normal range. Therefore, ABZ-loaded SLNs could be a potential new product for CE treatment.

Intraperitoneal administration of lidocaine or tramadol alone or in combination on postoperative pain after ovariohysterectomy in dogs.

The present prospective randomized experimental study aimed to assess the intraperitoneal (ip) administration of lidocaine or tramadol, alone or in combination, on postoperative pain management following ovariohysterectomy in dogs. Eighteen healthy female mixed-breed dogs, aged 1-2 years, weighed 16.7 ± 3.8 kg, were used. Animals were sedated with acepromazine (0.1 mg/kg, intramuscular). Forty minutes later, anaesthesia was induced through intravenous titration with diazepam (0.5 mg/kg) and ketamine (10 mg/kg) and maintained with isoflurane 1.5%. Afterwards, ovariohysterectomy was performed, and prior to the closure of the linea alba, animals received lidocaine containing epinephrine (8.8 mg/kg, ip) in group L, tramadol (4 mg/kg, ip) in group T and lidocaine containing epinephrine (8.8 mg/kg, ip) plus tramadol (4 mg/kg, ip) in the LT group. Cortisol, vital signs and pain scoring systems were evaluated at different time points. Vital signs did not change among the groups. Cortisol level in the LT group significantly decreased compared to the L and T groups one, three and six hours after surgery. Pain scores also did not change among the groups based on Sammarco and Simple descriptive (SDS) scoring method. However, pain scores in the LT group were higher than the two other groups according to the University of Melbourne pain scale (UMPS) and the short form of Glasgow pain scale (CMPS-SF). According to the obtained results, the combination of lidocaine and tramadol seemed to be able to provide better analgesia compared with their separate administration. Therefore, combined intraperitoneal administration of lidocaine (8.8 mg/kg) and tramadol (4 mg/kg) with a final volume of (0.2 ml/kg) following ovariohysterectomy is recommended.

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.

Comparative evaluation of therapeutic effects of silymarin and hydrocortisone on clinical and hematological alterations, and organ injury (liver and heart) in a low-dose canine lipopolysaccharide-induced sepsis model.

The present study aimed to examine the effectiveness of silymarin compared to hydrocortisone on clinical and hematological alterations and organ injury (liver and heart) in a low-dose canine lipopolysaccharide (LPS)-induced sepsis model. Fifteen clinically healthy dogs were randomly categorized into three equal groups: Two dogs in group A, LPS (0.10 µg kg-1, IV) was injected (control, n = 5); Group B was similar to group A, with the difference that silymarin bolus (10.00 mg kg-1, IV, once) was injected 40 min after LPS injection. Group C was similar to group B with the difference that hydrocortisone bolus (2.00 mg kg-1, IV, once) was administrated instead of silymarin. Five mL of blood was collected at baseline, 1, 3, and 6 hr of the study. Septic control dogs experienced a significant reduction in red blood cells count (RBC), hemoglobin (Hb), and hematocrit (HCT) and a significant elevation in serum activities of aspartate aminotransferase (AST), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), creatine kinase isoenzyme MB (CK-MB), and plasma cardiac troponin I (cTnI) concentration. We noticed a significant increase in RBCs, Hb, and HCT, and a significant decrease in AST, ALP, LDH, CK-MB, and cTnI in the silymarin group in comparison with hydrocortisone and control group. Our results suggested that silymarin had a positive influence on sepsis due to protecting RBCs, and decreasing organ (heart and liver) injury. These findings supported the hypothesis that silymarin could be more effective than routine corticosteroid therapy in sepsis.

The Effects of Propofol Anesthesia on Lipid Profile and Some Biochemical Indices in Cats.

The aim of the present study was to evaluate lipid profile (triglyceride, total cholesterol, HDL, LDL, and VLDL), pancreas (lipase and amylase), liver (AST, ALT, and ALP), blood urea nitrogen, creatinine, uric acid, sodium and potassium function indicators in cats undergoing two different durations of anesthesia with propofol. Ten adult female cats were randomly divided into two groups (n= 5) and anaesthetized with propofol 1% (induction: 8 mg/kg; infusion: 0.3 mg/kg/min) for either 45 or 90 minutes. Blood samples were collected at predetermined intervals up to 72 hours later. Comparison of the measured variables between treatments did not show significant differences. Triglyceride and cholesterol levels showed significant increase after induction of anesthesia (P < .05). The highest triglyceride and cholesterol values were recorded at 6 and 24 hours. HDL was lower while LDL and VLDL were higher at several time points after anesthesia (P < .05). Higher values of lipase, ALT and AST were detected after induction (P < .05). All the observed alterations were within normal ranges. In conclusion, propofol anesthesia was associated with some changes in lipid profile, as well as pancreatic and liver function indices, which should be considered in clinical situations. It seems that in the absence of pre- or co-existing disturbances, induction and maintenance of anesthesia with propofol did not carry additional risk to cats.

Clinical, hematologic, hemostatic, and serum biochemical findings related to survival in Arabian horses with colic.

BACKGROUND: The efficacy of clinical examinations, and hematologic, hemostatic, and serum biochemical assessments to determine appropriate early medical or surgical therapy in Arabian horses with colic have not been well-described. OBJECTIVES: We aimed to evaluate clinical and hematologic findings, coagulation-fibrinolytic parameters, and organ dysfunction markers in association with survival in horses with colic. METHODS: Seventeen Arabian horses with colic and ten healthy horses were enrolled. After clinical examinations, including heart rates (HRs), capillary refill times (CRTs), abdominal sounds, rectal palpations, and nasogastric (NG) reflux assessments in both healthy and diseased horses, blood samples were collected and hematologic parameters, coagulation indices such as the prothrombin time (PT), activated partial thromboplastin time (APTT), plasma d-dimer (PDD), platelet count, fibrinogen, and hepatorenal biomarkers were measured. Based on the outcomes, horses with colic were divided into survival (10) and nonsurvival (7) groups, and all mentioned parameters were compared with the control group horses. RESULTS: Unlike HRs, CRTs, and abdominal sounds, rectal palpations (57.1%) and NG reflux (28.6%) findings were least sensitive in being able to determine survival. At least three of the five coagulation indices were significantly abnormal (prolonged PT, increased PDD, and decreased plasma fibrinogen) in 71.4% of the horses in the nonsurvival group compared with the control and survival groups. Significantly increased urea, creatinine, and bilirubin levels, and AST activities were also observed in the survival group compared with the control and nonsurvival groups. CONCLUSIONS: The combination of prolonged PT, increased PDD, and hypofibrinogenemia could be a reliable predictor of survival in horses with colic. The significant renal and hepatic dysfunctions in nonsurviving horses with colic appeared to be consistent with disseminated intravascular coagulation (DIC) and a grave prognosis.

Serum Apelin-36 alteration in late pregnancy and early lactation of dairy cows and its association with negative energy balance markers.

The beneficial roles of Apelin on both energy metabolism and insulin sensitivity have been described in previous researches, but it has been little studied in dairy cows. The aim of the present study was to determine the serum Apelin-36 concentration in late pregnancy and early lactation in dairy cows and its association with negative energy balance markers. Thirty Holstein dairy cows (multiparous; n = 15 and primiparous; n = 15) with body condition score 3-3.75 at parturition were selected and blood samples were obtained for metabolic profile one month before and one month after parturition. Apelin-36, glucose, insulin, ß-hydroxybutyric acid (BHBA), non-esterified fatty acid (NEFA), cholesterol, triglyceride (TG) and high density lipoproteins (HDL) were measured using commercial kits. BCS and milk production were recorded during the study. There was no effect of parity on Apelin-36, cholesterol, TG, HDL, BHB and NEFA concentrations before lactation; while insulin and glucose levels were higher in primiparous cows than multiparous cows at this period. None of the factors showed any significant difference between multiparous and primiparous cows after lactation. Serum NEFA concentration were increased after parturition, while Apelin-36, insulin and glucose concentrations were decreased after parturition in primiparous and multiparous cows. Significant correlations were observed between serum Apelin and insulin (P = .041, r = 0.672), NEFA (P = .027, r = -0.808) and glucose (P = .037, r = 0.757). In conclusion, our results showed that serum Apelin-36 concentration decreased after parturition in dairy cow. Alteration of Apelin-36 secretion after parturition may represent an endocrine adaptation in dairy cow during the lactating period.

Occurrence and potential causative factors of immune-mediated hemolytic anemia in cattle and river buffaloes.

The main objectives of this study were to determine the occurrence and potential causative factors of Immune-mediated hemolytic anemia (IMHA) in native cattle and water buffaloes from southwest of Iran. Fifty-three anemic animals (37 cattle and 16 buffaloes) were studied. A full clinical history and physical examinations were undertaken for all animals. Four clinically healthy cattle and four healthy buffaloes were also used as control animals. Blood samples were subjected to a complete blood count, Coombs' test, erythrocyte osmotic fragility test and serum biochemical analysis. IMHA was diagnosed in 12 (32.43%) cattle and 6 (37.50%) buffaloes based on the Coombs' test. Underlying or concurrent diseases, including theileriosis, anaplasmosis, vaccination, and pneumonia were detected in 11 cattle and four buffaloes. Primary or idiopathic IMHA was identified in one cattle and two buffaloes that their Coombs' test was positive. Hematologic and biochemical findings in the cattle with IMHA included a nonregenerative anemia, leukopenia, thrombocytopenia, increased osmotic fragility, hyperbilirubinemia and elevated serum alkaline phosphatase, aspartate aminotransferase and lactate dehydrogenase activities. It can be concluded that IMHA occurs in a significant proportion of anemic cattle and river buffaloes in southwest of Iran. The occurrence of IMHA in both cattle and buffaloes is mostly secondary to infectious diseases especially theileriosis and anaplasmosis. Clarification of the mechanisms of primary or idiopathic and secondary IMHA in cattle and buffaloes require further studies.

In vitro antileishmanial activity of fisetin flavonoid via inhibition of glutathione biosynthesis and arginase activity in Leishmania infantum.

With the increasing emergence of drug resistant Leishmania sp. in recent years, combination therapy has been considered as a useful way to treat and control of Leishmaniasis. The present study was designed to evaluate the antileishmanial effects of the fisetin alone and combination of fisetin plus Meglumine antimoniate (Fi-MA) against Leishmania infantum. The IC50 values for fisetin were obtained 0.283 and 0.102 µM against promastigotes and amastigote forms, respectively. Meglumine antimoniate (MA, Glucantime) as control drug also revealed IC50 values of 0.247 and 0.105 µM for promastigotes and amastigotes of L. infantum, respectively. In order to determine the mode of action of fisetin and Meglumine antimoniate (MA, Glucantime), the activities of arginase (ARG), catalase (CAT), glutathione peroxidase (GPx) and superoxide dismutase (SOD) were measured. Moreover, intracellular glutathione (GSH) and nitric oxide (NO) levels in L. infantum-infected macrophages and L. infantum promastigotes which were treated with IC50 concentrations of fisetin, MA and Fi-MA were investigated. Our results showed that MA decreased CAT and SOD activity and increased NO levels in L. infantum-infected macrophages. In promastigotes, MA inhibited parasite SOD activity and reduced parasite NO production. The decreased levels of most of the antioxidant enzymes, accompanying by the raised level of NO in treated macrophages with MA, were observed to regain their normal profiles due to Fi-MA treatment. Furthermore, fisetin could prevent the growth of promastigotes by inhibition of ARG activity and reduction of GSH levels and NO production. In conclusion, these findings showed that fisetin improves MA side effects.

Amiodarone triggers induction of apoptosis in cutaneous leishmaniasis agents.

Leishmaniasis is a parasitic disease that is an important problem of public health worldwide. The aim of this study was to assess the cytotoxic effects of amiodarone (AMD) on Leishmania tropica (MHOM/AF/88/KK27) and L. major (MRHO/IR/75/ER) promastigotes and to observe the programmed cell death features. The colorimetric MTT assay was used to find L. tropica and L. major viability and the obtained results were expressed as 50% inhibitory concentration (IC50). Annexin-V FLUOS staining was performed to study the cell death properties of AMD using fluorescence-activated cell-sorting analysis. Qualitative analysis of the total genomic DNA fragmentation was performed by agarose gel electrophoresis. Furthermore, to observe changes in cell morphology, promastigotes were examined using light microscopy. The IC50 was achieved at 55 and 81 µM for L. tropica and L. major after 48 h of incubation, respectively. In both strains, AMD induced death with features of apoptosis, including externalization of phosphatidylserine, DNA laddering, and cell shrinkage. Our findings indicate that AMD may induce apoptosis on the causative agents of cutaneous leishmaniasis.

Detection of Sarcocystis spp. in cattle (Bos taurus) and water buffaloes (Bubalus bubalis) in Iran by PCR-RFLP.

Sarcocystis species are cyst-forming intracellular protozoan parasites. Cattle are mainly infected with Sarcocystis cruzi, Sarcocystis hominis and Sarcocystis hirsuta. Water buffaloes are intermediate hosts for Sarcocystis fusiformis, Sarcocystis levinei (S. cruzi-like species), Sarcocystis dubeyi, Sarcocystis sinensis (S. hominis-like species) and Sarcocystis buffalonis (S. hirsuta- like species). The aim of this study was Identification of Sarcocystis spp. in slaughtered cattle and water buffaloes in Ahvaz, Khuzestan province by PCR-restriction fragment length polymorphism. Meat inspection was done on 124 cattle and 147 water buffaloes. From each animal tissue samples (each 50 g) from heart, esophagus, diaphragm and intercostal muscle were collected during meat inspection. Samples examined with digestion method. Genomic DNA of 80 positive samples was extracted and their 18S rRNA gene was amplified. PCR products were digested by restricted enzymes (FokI, SspI and DraI). S. cruzi in cattle and S. fusiformis in water buffaloes were identified. Our study clarified that sarcocystosis in cattle in Ahvaz district may be results acute infection according to determined species, but in buffaloes as S. fusiformis was detected we may expect only economic loss follow up slaughterhouse inspection.

Designing and evaluation of Dot-ELISA for diagnosis of Fasciola infection in cattle.

Fasciolosis is a disease caused by liver fluck of the genus of Fasciola. Diagnosis of fasciolosis has been challenging for a long period due to low sensitivity of the coprological diagnostic method. In this study, an in-house Dot-ELISA method; using excretion-secretory (ES Ag) and Crude (Cr Ag) antigens of Fasciola was described for diagnosis of fasciolosis in cattle. For this purpose, the sera specimens of slaughtered cattle were taken and examined for Fasciola infection. Sera from two groups of cattle, one infected with Fasciola (n = 60) and the other non-infected with Fasciola (n = 60), were used in the Dot- ELISA test. All sera were tested and evaluated. Except specificity, other parameters such as, sensitivity, accuracy, positive and negative predictive values of Dot- ELISA with ES Ag were better than those of Dot- ELISA with Cr Ag. In conclusion, excretory-secretory antigen dependent Dot-ELISA can be used as a reliable sero-diagnostic test for Fasciola infection in cattle.

Seroprevalence of Toxoplasma gondii in water buffaloes (Bubalus bubalis) in South-West of Iran.

The prevalence of antibodies to Toxoplasma gondii was conducted in 300 buffaloes from Ahvaz, Kouzestan province, southwest of Iran. Blood sera were screened using a Modified agglutination test (MAT) incorporating 2-mercaptoethanol. Positive reactions in sera dilutions above 1:25 were considered as indicative for the presence of T. gondii antibodies. The overall prevalence of infection in the animals was 14.33% with titers of 1:25 in 21, 1:50 in 12, 1:100 in 6, 1:200 in 2 and 1:400 in 2. The prevalence was different in relation to the sex with buffaloes with 19.7% and 7% in females and males respectively. These results indicate that T. gondii infection in water buffaloes of Khouzestan is relatively high and consumption of buffalo meat may be a risk factor for humans in Ahvaz, southwest of Iran.

Occurrence of anti-Toxoplasma gondii antibodies in female cattle in south-west of Iran.

Toxoplasmosis, caused by Toxoplasma gondii, is a significant disease in livestock and humans. In Iran, studies shows that T. gondii infection in humans is relatively high and prevalence is associated mainly with consumption of undercooked meat or meat products. We have examined 450 serum samples from female cattle distributed over all Ahvaz, the center of Khouzestan province, south-west of Iran. IgG antibodies to T. gondii were assayed by the modified agglutination test using whole tachyzoites of T. gondii, and found in 71 (15.77%) of 450 cattle with titers of 1:25 in 38, 1:50 in 18, 1:100 in 11, 1:200 in three and 1:400 in one. Titers of antibodies were decreased in cattle over 2 years old. These results indicate that T. gondii infection in cattle of Khouzestan is relatively considerable, but not very high and consumption of beef may be a source of infection for humans in south-west of Iran.