G-CSF administration attenuates brain injury in rats following carbon monoxide poisoning via different mechanisms.

Acute severe carbon monoxide (CO) poisoning induces hypoxia that leads to cardiovascular and nervous systems disturbances. Different complex mechanisms lead to CO neurotoxicity including lipid peroxidation, inflammatory and immune-mediated reactions, myelin degeneration and finally neuronal apoptosis and necrosis. Granulocyte colony-stimulating factor (G-CSF) is considered to be a novel neuroprotective agent. In this study, we evaluated the efficacy of G-CSF therapy on CO neurotoxicity in rats with acute CO poisoning. Rats were exposed to 3000 ppm CO in air (0.3%) for 1 h, and then different doses (50,100, and 150 µg/kg) of G-CSF or normal saline were administrated intraperitoneally. Water content of brain as an indicator for total edema and blood brain barrier integrity (Evans blue extravasation) were evaluated. Malondialydehyde was determined in order to evaluate the effect of G-CSF on CO-induced lipid peroxidation in brain tissues. Also, the effect of G-CSF on myeloperoxidase activity in the brain tissue was evaluated. The effect of G-CSF administration on induced apoptosis in the brain was measured using TUNEL method. To evaluate the level of MBP, STAT3 and pSTAT3 and HO-1 proteins and the effect of G-CSF on these proteins Western blotting was carried out. G-CSF reduced water content of the edematous poisoned brains (100 µg/kg) and BBB permeability (100 and 150 µg/kg) (P < 0.05). G-CSF (150 µg/kg) reduced the MDA level in the brain tissues (P < 0.05 as compared to CO poisoned animals). G-CSF did not decrease the MPO activity after CO poisoning in any doses. G-CSF significantly reduced the number of apoptotic neurons and Caspase 3 protein levels in the brain. Western blotting results showed that G-CSF treatment enhanced expression of HO-1 and MBP, STAT3 and pSTAT3 proteins in the brain tissues. Based on our results, a single dose of G-CSF immediately after CO poisoning significantly attenuates CO neurotoxicity via different mechanisms. © 2015 Wiley Periodicals, Inc. Environ Toxicol 32: 37-47, 2017.

An experimental ovine Theileriosis: The effect of Theileria lestoquardi infection on cardiovascular system in sheep.

The malignant ovine theileriosis is caused by Theileria lestoquardi, which is highly pathogenic in sheep. Theileriosis involves different organs in ruminants, but the effect of the disease on the cardiovascular system is unclear. To understand the pathogenesis of T. lestoquardi on the cardiovascular system, Baluchi breed sheep were infected with the mentioned parasite by releasing unfed adults of Hyalomma anatolicum anatolicum, which were infected with T. lestoquardi. The infected sheep were clinically examined on days 0, 2, 5, 7, 10, 12, 14, 17, and 21, and the blood samples were collected for biochemical parameters measurement. At termination of the experiment, the infected sheep were euthanized and pathological examinations of heart tissue were conducted. During experimental infection of sheep with T. lestoquardi, activities of cardiac troponin I (cTnI), lactate dehydrogenase, and aspartate aminotransferase, were significantly increased (P˂0.05), while a conspicuous decrease (P˂0.05) was observed in creatine phosphokinase activities. Alterations made in biochemical factors almost coincided with the presence of piroplasm in the blood and schizont in lymph nodes. Maximum and minimum of parasitemia in the sheep stood between 3.3% and 0.28%, respectively. In addition, electrocardiography revealed sinus tachycardia, sinus arrhythmia, sino-atrial block and ST-elevation, atrial premature beat, and alteration in QRS and in T waves' amplitude. Heart histopathological examination showed hyperemia, infiltration of mononuclear inflammatory cells into interstitial tissue, endocarditis, and focal necrosis of cardiac muscle cells. In addition, in one of the sheep, definite occurrence of infarction was observed. The results indicate that T. lestoquardi infection has devastating pathological impacts on the cardiovascular system of sheep. Furthermore, measurement of the cTnI amount is a useful biochemical factor for diagnosis and for better understanding of the severity and progression of the disease and its effects on cardiac tissue.

Outbreak of ovine congenital toxoplasmosis in Iran, confirmed by different diagnostic methods.

Toxoplasma gondii is a zoonotic obligatory intracellular protozoan parasite with the capability to infect all warm-blooded animals. One of the great concerns is that it can lead to ovine abortion in sheep growing industry. Different diagnostic methods such as serology, pathology, immunohistochemistry, bioassay and molecular detection have been used in order to detect ovine abortion associated with T. gondii. In this case, an outbreak of congenital toxoplasmosis based on serological, macroscopic, pathological detection and isolation of T. gondii by bioassay is described and emphasized on the importance of this route of transmission that caused lamb losses and an increase in possible sources of infection for human and environment.

Protective and hypoglycemic effects of celery seed on streptozotocin-induced diabetic rats: experimental and histopathological evaluation.

AIMS: Diabetes mellitus is a major manifestation of metabolic disorder which presents with hyperglycemia (high levels of serum blood sugar). In the present study, we aimed to investigate the effects of celery seed extract on different biochemical factors and histopathological changes in normal and streptozotocin (STZ)-induced diabetic rats. METHODS: A total of 35 male Wistar rats were divided into five groups (one normal and four diabetic groups). STZ was injected intraperitoneally to induce diabetes. The effects of hexane extract of celery seed and glibenclamide (as a positive control) were compared. Blood samples were analyzed on days 0, 18, and 33, and histopathological evaluations were performed at the end of the study. RESULTS: Glucose, triglycerides, and cholesterol levels significantly decreased, whereas insulin and high-density lipoprotein (HDL) levels increased in the extract-administered groups, as compared to the negative diabetic control group (P < 0.0001). The concentrations of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum of the extract-administered groups were significantly less than the negative control group (P < 0.0001). Histopathological reports revealed significantly less atrophy, necrosis, and inflammation in the rats receiving celery seed extract compared to the negative control group. CONCLUSIONS: The results indicated that celery seed extract can be effective in controlling hyperglycemia and hyperlipidemia in diabetic rats, and demonstrated its protective effects against pancreatic toxicity resulting from STZ-induction.

Genotyping of Clostridium perfringens isolated from healthy and diseased ostriches (Struthio camelus).

BACKGROUND AND OBJECTIVES: Clostridium perfringens is more prevalent type of clostridia genus isolated from the intestinal tract of ostrich (Struthio camelus). Necrotic enteritis (NE) is a potentially fatal gastrointestinal (GI) disease of poultry and other avian species, which produces marked destruction of intestinal lining in digestive tract caused by C. perfringens. Pathogenicity and lesions are correlated with the toxins produced, thus toxin typing of the bacterium has diagnostic and epidemiological significance. The aims of the present study were to determine the biotypes of C. perfringens among ostrich's farms either diseased and healthy ones and to screen the isolates for major toxin genes (cpa, cpb, etx, and iA, cpb2, and cpe). MATERIALS AND METHODS: Thirty isolates of C. perfringens were obtained from NE-positive and NE-negative ostrich flocks in Khorasan-e-Razavi porvince and analyzed by multiplex PCR assay. RESULTS: All isolates were positive for alpha toxin gene (cpa) and five of those were positive for beta toxin gene (cpb). The presence of cpb2 gene was detected in a high percentage of isolates originating from both healthy (93.3%) and diseased flocks (80%). None of the isolate carried enterotoxin gene (cpe). CONCLUSION: The results suggest that types A and C of C. perfringens are the most prevalent types in ostrich in Iran. Due to detection of beta2 toxin gene in isolates from both healthy and diseased birds, it appears that the presence of cpb2 is not considered a risk by itself.

Clinical, laboratory and pathological findings in sub-acute monensin intoxication in goats.

Toxic effects of monensin, a polyether antibiotic mainly used as a coccidiostat, have been described in a wide range of animals. The present study was performed to investigate the toxic effects of monensin in goats. Seven adult goats were administered sodium monensin, 13.5 mg kg(-1), daily for five consecutive days via gastric gavage. Monensin toxicity was evaluated by clinical signs, serum biochemistry and pathology. Monensin exposure caused diarrhea, tachycardia and reduction in ruminal movements and body temperature. Significant increase of creatine kinase, lactate dehydrogenase, aspartate aminotransferase, total bilirubin, blood urea nitrogen, creatinine and erythrocyte superoxide dismutase were observed in monensin exposed goats. Reduction of erythrocyte glutathione peroxidase and elevation of serum malondialdehyde and troponin I were inconsistent. In necropsy, there were effusions in body cavities, vacuolar degeneration and coagulative necrosis in cardiac and skeletal muscles and renal tubular necrosis. These findings suggested that monensin intoxication in goats leads to cardiac, skeletal and renal damage and a wide range of biochemical abnormalities. Oxidative stress may be involved in the pathogenesis of monensin poisoning.

Crocetin attenuates spatial learning dysfunction and hippocampal injury in a model of vascular dementia.

Chronic cerebral hypoperfusion produces brain ischemia and is associated with related memory impairment. In this study, we prepared crocetin from crocin (the main water soluble pigment in saffron stigmas) in an acidic hydrolysis and its purity was evaluated using spectrophotometry, thin layer chromatography (TLC) and high performance liquid chromatography (HPLC). The memory enhancing effect of crocetin was then studied in a rat model of chronic cerebral hypoperfusion. Different doses of crocetin were administered intraperitoneally (i.p.). The spatial learning and memory function were tested using Morris water maze, after permanent occlusion of common carotids. Finally, the animals were euthanatized in CO2 chamber and histopathological changes in cerebral cortex and hippocampus were investigated. The results indicated that the escape latency time significantly decreased in crocetin treated rats, in comparison with control animals. Also, the percentage of time spent and traveled distance in target quadrant on final test trial day increased in crocetin 8 mg/kg group, compared to control group, while no difference was observed between groups in swimming speed. All behavioral results were confirmed by histopathological analysis. According to our data, crocetin (8 mg/kg) could properly protect cerebrocortical and hippocampus neurons against ischemia. In conclusion, treatment with crocetin could effectively prevent neuropathological alterations in hippocampus thereby resulting in improvement of spatial learning memory in rats after chronic cerebral hypoperfusion.

Protective effect of crocin on diazinon induced cardiotoxicity in rats in subchronic exposure.

UNLABELLED: This study was designed to evaluate the effectiveness of crocin, main component of Crocus sativus L. (Saffron) against subchronic diazinon (DZN) induced cardiotoxicity in rats. METHODS: Rats were divided into 7 groups; control (corn oil, gavage), DZN (15 mg/kg/day, gavage,), crocin (12.5, 25 or 50 mg/kg/day, i.p) plus DZN, vitamin E (200 IU/kg, i.p, three times per week) plus DZN and crocin (50 mg/kg/day, i.p) groups. Treatments were continued for 4 weeks. Creatine phosphokinase MB (CK-MB), malondealdehyde (MDA) and glutathione (GSH) levels were evaluated in heart tissue at the end of treatments. Levels of apoptotic proteins (Bax, Bcl2, caspase 3) and cytosolic cytochrome c were analyzed by Western blotting. Transcript levels of Bax and Bcl2 were also determined using qRT PCR. RESULTS: DZN induced histophatological damages and elevated the level of cardiac marker CK-MB. These effects were associated with increased MDA level, lower level of reduced GSH and induction of apoptosis through elevation of Bax/Bcl2 ratio (both protein and mRNA levels), cytochrome c release to the cytosol and activation caspase 3 in cardiac tissue. Crocin (25 and 50 mg/kg) or vitamin E improved histopathological damages, decreased MDA and CK-MB, increased GSH content and attenuated the increase of Bax/Bcl2 ratio, activation of caspase 3 and release of cytochrome c to the cytosol induced by DZN. In summary, DZN induced mitochondrial-mediated apoptosis in heart tissue of rat following subchronic exposure. Crocin, as an antioxidant, showed protective effects against DZN cardiotoxicity by reducing lipid peroxidation and alleviating apoptosis.

The effect of granulocyte colony-stimulating factor administration on carbon monoxide neurotoxicity in rats.

Granulocyte colony-stimulating factor (G-CSF) is considered to be a novel neuroprotective agent. Beneficial effects have been demonstrated by administrating G-CSF in different experimental stroke models. In this study, we evaluated the efficacy of G-CSF therapy on carbon monoxide (CO) neurotoxicity in rats exposed to acute CO poisoning. Immediately after exposure to 3,000 ppm of CO for 60 minutes, 50, 100, and 150 µg/kg of G-CSF or normal saline were administered to rats. Rats were sacrificed after 24 hours for serum marker analysis or 1 week for histopathological examination. Brain sections were stained with hematoxylin and eosin to assess leukocyte infiltration and hippocampal injury and with Luxol fast blue to assess demyelination. S100ß and glial fibrillary acidic protein (GFAP) serum levels were evaluated by commercial enzyme-linked immunosorbent assay kits. According to histopathological findings, G-CSF administration significantly restricted white-matter demyelination (150 µg/kg) (P = 0.006). Also, serum levels of S100ß in G-CSF-treated groups (100 and 150 µg/kg) decreased significantly (P < 0.01 and P < 0.05, respectively). In all does, G-CSF significantly reduced serum levels of GFAP (P < 0.01 for 50 µg/kg and P < 0.001 for other doses). Administration of G-CSF after CO poisoning attenuates brain cell damage through remyelination. G-CSF also decreases levels of related biomarkers, such as S100ß and GFAP.

Serum troponin I as an indicator of myocarditis in lambs affected with foot and mouth disease.

Foot and mouth disease (FMD) is a highly contagious viral disease of cloven-hooved livestock and wildlife results to relatively high mortality in young animals. Despite the numerous reports of FMD-related death in neonates, there is little data available on various aspects of FMD in lambs. This report describes myocarditis associated with FMD in five, one week to three months old lambs. The lambs were depressed and afebrile and two of lambs showed foamy salivation associated with shallow ulcers in oral cavity. Electrocardiography (ECG) revealed sinus tachycardia, multifocal ventricular premature beats and ventricular fibrillation. Serum biochemistry showed high levels of troponin I concentration and CK and AST activity. In Pathology, there were multiple pale areas in the subepicardial and subendocardial muscles and; widespread degeneration and coagulative necrosis of myocardium. The serum troponin I assay and ECG can be used for diagnosis of myocarditis and prognosis of affected lambs during FMD outbreak.

Survey of dermatological conditions in a population of domestic dogs in Mashhad, northeast of Iran (2007-2011).

university small animal clinic, 1299 Iranian domestic dogs presented from September 2007 through March 2011 to the Ferdowsi University of Mashhad Veterinary Teaching Hospital, were examined. Dermatological disorders were noted in 17.00% (221/1299) of all the dogs examined. Pruritus was the most common presenting sign, accounting for 25.35% of the dermatological consultations. It was followed by erythema, maculo-papular-pustular eruptions (16.97%), erosive or ulcerative lesions (16.74%), scaling or crusting (13.02%), alopecia (8.84%) and visible ectoparasites (7.44%). The most common primary final diagnoses were superficial pyoderma, cutaneous manifestations of canine leishmaniasis, flea infestation and allergy, tick infestation, atopic dermatitis, scabies, unspecified dermatoses, otitis, furunculosis and food allergy. There were no apparent age or sex predilections for dermatological disease as a whole. Spitz (odds ratio = 3.38; p = 0.001), Terriers (odds ratio = 2.52; p < 0.001) and German Shepherds (odds ratio = 1.90; p = 0.001) appeared to be at increased risk for dermatological disease. In addition, Khorasani large cross breed dogs (odds ratio = 0.36; p = 0.003) and mixed breed dogs (odds ratio = 0.33; p < 0.001) showed decreased risk for dermatological conditions. To the best of our knowledge, this is the first survey study on canine dermatological conditions carried out in Iran.

Acute oxalate intoxication associated to ingestion of eshnan (Seidlitzia rosmarinus) in sheep.

An outbreak of acute oxalate intoxication in a sheep flock was associated to Seidlitzia rosmarinus (Chenopodiaceae) with a mortality rate of about 19%. Affected sheep showed marked azotemia and hypocalcemia. Post-mortem findings included congestion and hemorrhage in visceral organs, ruminitis frequently associated with precipitation of birefringent calcium oxalate crystals, and acute nephrosis with numerous birefringent calcium oxalate crystals in renal tubules. This is the first report of oxalate poisoning due to ingestion of S. rosmarinus in sheep.

The first report of Hepatozoon canis infection of a dog in Iran.

An 11-year-old male dog was presented with a 1-week history of inappetence, weight loss and hind limb paralysis. Physical examination revealed weakness, depression, incoordination of the posterior limbs, peripheral lymphadenopathy and pale mucous membranes. Laboratory analysis of blood samples revealed anaemia, thrombocytopenia and low serum albumin concentration. The diagnosis was confirmed microscopically, by demonstrating the presence of Hepatozoon canis gametocytes within neutrophils in Giemsa-stained peripheral blood smears and bone marrow smear. Also, schizonts of H. canis were seen in tissue sections of muscles, lymph nodes, spleen and liver. To the best of authors' knowledge, this is the first description of H. canis infection in a dog in Iran.