Protective role of silymarin and D-penicillamine against lead-induced liver toxicity and oxidative stress.

This study was performed to assess hepatotoxicity and alterations in liver antioxidant defence in acute lead (Pb) exposure and the protective effects of silymarin in comparison to D-penicillamine in rats. Forty eight Albino rats were divided in eight groups and received the following treatments in a 10-day experiment - group 1: normal saline as control; group 2: 25-mg/kg Pb acetate, intraperitoneally (IP) for the last 5 days; group 3: 100-mg/kg D-penicillamine, IP for the last 5 days; group 4: 200-mg/kg silymarin, orally for 10 days; and groups 5, 6, 7 and 8: in addition to Pb, they received D-penicillamine, for the last 5 days, silymarin for 10 days, a combination of silymarin for 10 days and D-penicillamine for the last 5 days and silymarin for the last 5 days, respectively. Pb acetate exposure induced significant elevation in serum alkaline phosphatase (ALP), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) enzyme activities in group 2 compared to control group. Significant reductions in serum total protein and albumin in all Pb-exposed groups and in serum glucose in groups 2, 6 and 8 were also observed. Liver tissue superoxide dismutase and glutathione peroxidase were significantly lower in groups 2 and 8 compared to control group. Silymarin pretreatment and D-penicillamine administration in groups 5, 7 and 8 could significantly lower ALP, ALT and AST and improve liver antioxidant enzymes. Thus, acute Pb exposure induced hepatotoxicity with suppression of liver antioxidant defence system and silymarin, as an antioxidant could alleviate this effect.

Evaluation of oxidant/antioxidant status, trace mineral levels, and erythrocyte osmotic fragility in goats naturally infected with Anaplasma ovis.

Anaplasma ovis, an arthropod-borne pathogen that infects erythrocytes, is the major cause of ovine and caprine anaplasmosis. This study was performed to assess in goats infected with A. ovis the osmotic fragility of erythrocytes, antioxidant status, and serum levels of microminerals. Blood samples were collected from 104 mixed breed goats in Ahvaz area, southwest Iran and subjected to parasitologic, hematologic, oxidant/antioxidant, and micromineral assessment. Anaplasma infection was detected in 30 samples (28.8 %) by microscopic examination of blood smears while PCR-RFLP analysis revealed infection with A. ovis in 68 samples (65.4 %). Studied animals were divided into three groups based on A. ovis infection: Uninfected goats as control group (group 1), PCR positive without parasitemia (group 2) and PCR positive with parasitemia (group 3). Hematological evaluation showed significantly increased lymphocyte and monocyte counts in Anaplasma-infected groups (group 2 and 3). A significantly lower MCHC and higher MCV were also observed in infected groups. In group 3 significant rises in erythrocyte's osmotic fragility in different salt concentrations and also in median corpuscular fragility (MCF) was seen. Evaluation of the antioxidant defense system of the erythrocytes revealed a decrease in total antioxidant capacity (TAC) and superoxide dismutase (SOD) activity in group 3. There was no significant difference in serum micromineral levels between infected and uninfected animals. Overall, the observed substantial decrease in the antioxidant enzyme activities with remarkable elevated levels of erythrocyte osmotic fragility indicate high exposure of erythrocytes to oxidative damage in Anaplasma-infected goats. These results also suggest that the disturbed antioxidant defense mechanisms in caprine anaplasmosis can promote the development of anemia.

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.

Evaluation of intraperitoneal administration of morphine on post-operative pain management after ovariohysterectomy in dogs.

The present prospective randomized experimental study was designed to assess pain control with intraperitoneal morphine following ovariohysterectomy in dogs. A group of 12 mixed breed female dogs, aged 1-2 years, weighing 19.95 ± 0.95 kg were included. Forty minutes after sedation with 0.05 mg/kg intramuscular acepromazine 1%, anaesthesia was induced with propofol (4 mg/kg). The dogs were connected to the inhalation anaesthesia circuit using isoflurane. Ovariohysterectomy was performed, and before the closure of linea alba, the animals received intraperitoneal morphine (0.5 mg/kg) (in group M) and saline (0.2 ml/kg) (in group S). No significant difference was detected in total protein and glucose levels between the groups, while the cortisol level in group M was significantly lower than group S 1, 3 and 6 h after surgery. Furthermore, the comparison of the rectal temperature, heart rates and respiratory rates showed no major differences. Additionally, no significant alterations were detected between the groups considering the changes in the pain scores with simple descriptive score, Glasgow, University of Melbourne pain scale, sedation status and Sammarco methods. Finally, three cases in group S and two cases in group M were given an intramuscular analgesic rescue dose of morphine. Although a significant decline was observed in cortisol levels following intraperitoneal morphine administration, there were no beneficial changes in the efficiency of post-operative analgesia in status and clinical signs compared to the control group. Further studies are required to investigate intraperitoneal morphine effectiveness in post-operative pain management.

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.

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.

Detection of Theileria lestoquardi cross infection in cattle with clinical theileriosis in Iran.

Theileriosis caused by Theileria lestoquardi (malignant ovine theileriosis) in sheep and Theileria annulata (tropical theileriosis) in cattle is an important hemoprotozoal tick-borne disease in Iran. Due to major biologic and phylogenic similarities of these two species, this study was carried out to investigate the occurrence of natural infections with T.lestoquardi and T.annulata in cattle with clinical theileriosis in Ahvaz, southwest Iran. Fifty one cattle were selected based on clinical signs of theileriosis and confirmation by microscopic examination of blood smears. Blood samples were collected from each animal and hematologic and microscopic examinations were performed. Theileria piroplasmic forms were detected in all affected cattle. Pale mucous membranes (43.14%), icterus (11.76%) and fever (70.6%) were also observed. PCR-RFLP analysis revealed T. annulata infection in all tested cattle while coinfections with T. lestoquardi were found in two samples (3.92%). All sampled cattle including the two with mixed species Theileria infection were anemic. This is the first report of Theileria species cross infections in cattle with clinical theileriosis in Iran. It can be concluded that cattle can be infected with both pathogenic Theileria species, T. lestoquardi and T. annulata which can be an important issue in the epidemiology and spread of ovine malignant theileriosis.

Molecular Detection and Identification of Theileria Species by PCR-RFLP Method in Sheep from Ahvaz, Southern Iran.

BACKGROUND: The present study was carried out to investigate the accurate status of ovine Theileria infection in sheep from Ahvaz and surrounding region, a tropical area southwest Iran. METHODS: A PCR-RFLP method based on 18S ribosomal RNA gene was designed which could detect and differentiate Theileria and Babesia spp. and also differentiate main Theileria species in sheep at the same time. 119 sheep blood samples were collected from Ahvaz and surroundings. RESULTS: Microscopic examination of blood smears revealed 69.7% (83/119) infection with Theileria spp. Of the total samples subjected to PCR, 89% (106/119) were found to be positive, all of which were identified as Theileria by RFLP analysis using enzyme Hind II. In enzymatic digestion of PCR products by Vsp I, 91.5% (97/106) of Theileria positive samples were identified as T. ovis while mixed Theileria infections were found in 9 samples. The samples with mixed infections were analyzed with an additional nested PCR-RFLP method, by HpaII enzyme digestion. 3 samples with T. lestoquardi infection, 1 sample with T. ovis and T. annulata, 1 sample with T. lestoquardi and T. annulata, and 4 samples with T. ovis, T. lestoquardi and T. annulata mixed infections were detected. CONCLUSION: Ovine theileriosis caused by T. ovis is highly prevalent in southwest Iran while T. lestoquardi and T. annulata infection can be detected in a lesser propor-tion of sheep in this region. The new PCR-RFLP method that was designed in this study, can serve as a beneficial diagnostic tool, especially in T. ovis prevalent re-gions.

Antioxidant effect of different vitamins on methemoglobin production: An in vitro study.

Nitrite intoxication occurs frequently in ruminants and equines. The most common treatment of this disorder is administration of 1% methylene blue, although the use of some antioxidant agents e.g. vitamins and complementary treatment may also be useful. The aim of this study was to evaluate the in vitro antioxidative effects of some vitamins on methemoglobinemia induced by sodium nitrite. For this purpose the blood sample of a healthy dairy cattle was pre-incubated with three different concentrations (5, 10, 20 mmol L(-1)) of each vitamin (E, C, B1, A and a combination of vitamin E and vitamin C) as antioxidant agent at 4 (°)C for 24 hours. A control group with normal saline instead of vitamin was applied. Then, all samples were treated with sodium nitrite (10 mmol L(-1)) as an oxidant agent for 10 minutes and the level of methemoglobin formation was measured spectrophoto-metrically. The results revealed that the level of methemoglobin decreased significantly (P < 0.05), when vitamin E (10 and 20 mmol L(-1)) and vitamin C (5 mmol L(-1)) was applied to the tests, separately. Vitamin C at the concentration of 20 mmol L(-1), was not effective, but it even increased methemoglobin formation significantly. Combination of vitamin E and C was significantly effective at concentration 5 mmol L(-1), but not at concentration 10 and 20 mmol L(-1). Vitamin A and vitamin B1 were not effective in any concentration. It was concluded that vitamins especially vitamin C and E can reduce oxidative effects which induced methemoglobin formation in vitro and could be used as an alternative medication.