Cucumis callosus (Rottl.) Cogn. fruit extract ameliorates calcium oxalate urolithiasis in ethylene glycol induced hyperoxaluric Rat model.

Cucumis callosus dry fruits are traditionally used as folk remedy to treat conditions like urethral irritations, urine stoppage or dribbling and other urinary ailments of man in north-west India. But no study is reported to validate this ethnic practice of using Cucumis fruit in urolithiasis. To evaluate anti-urolithiatic potential of Cucumis, hyperoxaluria was induced in rats by supplying 0.75% ethylene glycol (EG) + 1% ammonium chloride (AC) in drinking water for 14 days. Anti-urolithiatic activity of Cucumis callosus hydro-ethanolic extract (CCHEE) was assessed by measuring blood and urine biochemical parameters, oxidative stress indices, histopathology and osteopontin (OPN) expression. Administration of EG-AC to rats caused hyperoxaluria, crystalluria, azotaemia, oxidant/antioxidant imbalance (increase in lipid peroxidation (LPO), and decrease in glutathione (GSH) and catalase (CAT)), up-regulation of OPN and calcium oxalate (CaOx) crystal deposition in kidney. Treatment of afflicted rats with Cucumis fruits extract restored renal function to a great extent (CCHEE group), testified by improvement of stated parameters. Findings demonstrate curative efficacy of Cucumis fruit extract in EG induced urolithiasis of rats. The restoration of renal function was possibly by regulating renal stone formation via reducing urinary oxalate excretion, correcting oxidant/antioxidant imbalances, and reduced expression of OPN. Hence, results of this study validate the ethnic practice of using Cucumis fruit and conclude that fruit extracts have beneficial effects on CaOx urolithiasis and renal function.

Effect of plastic foreign body impaction on rumen function and heavy metal concentrations in various body fluids and tissues of buffaloes.

Rumen impaction by plastic waste (PW) is a common condition in the developing countries where ruminants are reared under extensive grazing. As heavy metals (HM) are used in the manufacture of plastic, we hypothesized that buffaloes with PW impaction would have suboptimal rumen function and higher content of HM in the rumen fluid, sera and body tissues. Buffaloes at an abattoir were categorized into impacted with PW or not (Group C and B, respectively; n = 30/group). From the animals of group C and B, rumen fluid and sera were collected ante-mortem, while muscle, liver and kidney were collected after exsanguination. Blood and rumen fluid of stall-fed buffaloes (Group A; n = 15) served as known negative control. Rumen function was assessed by protozoal density, motility, pH, methylene blue reduction time (MBRT) and sedimentation activity time (SAT). Concentration of HM such as mercury (Hg), lead (Pb), cadmium (Cd), chromium (Cr) and copper (Cu) in the body fluids, tissues and impacted PW was estimated by atomic absorption spectrometry (AAS). A significant decrease in the rumen protozoal density, motility and increase (P < 0.05) in the rumen fluid pH, MBRT and SAT were observed in group C as compared to group A and B. Concentration of Pb, Hg and Cd showed a moderate to strong significant negative correlation with rumen function indicators. Quantitatively, Cu content (ppm) was highest in PW with 0.9 ± 0.04. The concentration of HM in the body fluids and tissues was significantly (P < 0.05) elevated in group C, but was below the toxic levels. In stall-fed buffaloes, Hg, Pb and Cd were undetectable in body fluids. Collectively, the results indicate that long term exposure of buffaloes to HM from PW affect the health, productivity and increase the risk of entering the food chain.

Urokinase alteration and its correlation with disease severity and oxidative/nitrosative stress in buffalo calves with obstructive urolithiasis.

The present study was aimed to understand the development of bovine urolithiasis through measuring oxidative/nitrosative, cortisol and urokinase status and their relationship with disease severity. The cases of buffalo calves with obstructive urolithiasis were selected based on clinical signs, ultrasonography and laboratory examination of blood and urine (creatinine, urea nitrogen). Total 35 urolithiatic buffalo calves (urolithiatic controls) and 6 healthy calves (healthy controls) were used for study. Further, calves of urolithiatic control were sub-divided into two groups based on disease severity: mild (n = 10) and severe (n = 25) form. Oxidative/nitrosative stress were evaluated based on serum malondialdehyde (MDA), glutathione-S-transferase (GST), nitric oxide (NO) parameters. Serum cortisol was evaluated to measure stress hormone status. Serum and urine urokinase were measured and its relationship with disease severity and oxidative/nitrosative stress were established. Obstructive urolithiasis resulted in significant (p < .05) increase in biochemical parameters (creatinine, urea nitrogen), oxidant/antioxidant imbalance (increased MDA, and increased GST), nitrosative stress (increased nitric oxide), upregulated stress hormone (cortisol) in serum and elevated urokinase in serum and urine (p < .05) as compared to healthy controls. In non-parametric Kendall rank correlation (p < .01), a positive correlation was established between urokinase level and disease severity (urolithiasis). It is concluded that in bovine obstructive urolithiasis, increased oxidative/nitrosative stress, cortisol and urokinase play a significant role. The urokinase can help to understand pathophysiology of bovine urolithiasis because of having positive correlation with disease severity (urolithiasis) and stress markers.

Circulating oxidative stress caused by Psoroptes natalensis infestation in Indian water buffaloes.

The present study reports the circulating oxidative stress associated with Psoroptes natalensis infestation in Indian water buffaloes. Three non-descriptive water buffaloes, age ranging between 4 and 9 years, presented to Referral Veterinary Polyclinic, IVRI, for treatment served as clinical subject. The infested animals were treated with Ivermectin subcutaneously and Amitraz topically along with antioxidant like ascorbic acid, Vitamin E and selenium. The level of lipid peroxidase was significantly higher (3.94 ± 0.34) in Psoroptes infested buffalo and was reduced significantly (P ≤ 0.05) after treatment (1.56 ± 0.40). The significantly higher levels of MDA before treatment signify the role of lipid peroxide mediated skin lesions in P. natalensis infested buffaloes. Similarly the activities of the body antioxidant like GSH and CAT were significantly higher (P ≤ 0.05) after treatment. The less level of the body antioxidant (GSH) and reduced activities of the antioxidant enzymes like CAT and SOD before treatment imply that Psoroptes mite-infested buffaloes were in a state of significant oxidative stress. The study provides information on oxidative stress indices in P. natalensis infested buffaloes and gives additional insight regarding the pathogenesis of the disease and its management.

Depressed polymorphonuclear cell functions in periparturient cows that develop postpartum reproductive diseases.

The study was planned to see if there is any important and significant changes in the PMN function in cows suffering from postpartum reproductive diseases (PRD). Blood sampling was done from 41 pregnant cows on 15 days prepartum (-15d), calving day (0d), 15 days (15d) and 30 days (30d) postpartum and thorough gynaecological examination was performed on 0d, 15d, 30d and 45d for diagnosis of PRD like retained placenta (RP), clinical metritis (CM), clinical endometritis (CE) and delayed involution of uterus (DIU). The heparinised blood was used for isolation of PMN leukocytes for estimation of superoxide (SO), hydrogen peroxide (H2O2) and enzyme myeloperoxidase (MPO) activity in each group of cows. The SO production (ΔOD) was greater for normal (0.19 ± 0.05) than cows suffering from RP (-0.12 ± 0.09), CM (-0.15 ± 0.13) and CE (-0.07 ± 0.05) at -15d. The mean value was greater for normal cows (0.12) than the cows with PRD (0.05 to 0.9) at 30d. The H2O2 production was greater for normal than cows with PRD at all sampling days and significantly greater than cows with RP and CE at 15d (p < 0.01) and 30d (P < 0.05). The MPO activity (µmol/1 × 107) was greater for normal (18.77 ± 1.27) than for RP (12.52 ± 2.57) and CM (11.31 ± 3.30) cows on 0d. The depressed capability of the PMN from the cows with PRD to produce SO, H2O2 and MPO during the periparturient period indicated their association with the development of RP, CM and CE.

Clinicobiochemical changes in ivermectin toxicity in Doberman pinscher pups.

This study is based on naturally intoxicated Doberman pinscher pups. Six Doberman pinscher pups were overdosed 50 times of recommended ivermectin dose by the pet owner and presented to Referral Veterinary Polyclinic section of this institute in a state of coma. The pups were unable to hold their head up and had no pupillary reflex. The pups were examined clinically and samples of blood were taken for haemato-biochemical studies, before and after treatment. During intoxication there was marked haemoconcentration, increase in values of serum creatinine phosphokinase, gamma glutamyl transpeptidase but decrease in serum glucose levels. These pups were treated with isotonic dextrose normal saline, 1.0 mg/kg physostigmine and 3 mg/kg dexamethasone. Clinical improvement occurred in remaining 2 pups after 12 h of treatment. These pups were able to take a liquid diet after 3 day and became clinically normal after 2 week. The serum biochemical values gradually reached to normal in recovered pups after 2 weeks of treatment. It took a month for complete recovery from depression and blindness.

Antiurolithiatic and antioxidant efficacy of Musa paradisiaca pseudostem on ethylene glycol-induced nephrolithiasis in rat.

OBJECTIVE: Musa paradisiaca has been used in the treatment of urolithiasis by the rural people in South India. Therefore, we plan to evaluate its efficacy and possible mechanism of antiurolithiatic effect to rationalize its medicinal use. MATERIALS AND METHODS: Urolithiasis was induced in hyperoxaluric rat model by giving 0.75% ethylene glycol (EG) for 28 days along with 1% ammonium chloride (AC) for the first 14 days. Antiurolithiatic effect of aqueous-ethanol extract of M. paradisiaca pseudostem (MUSA) was evaluated based on urine and serum biochemistry, microscopy of urine, oxidative/nitrosative indices, kidney calcium content, and histopathology. RESULTS: Administration of EG and AC resulted in increased crystalluria and oxaluria, hypercalciuria, polyuria, crystal deposition in urine, raised serum urea, and creatinine as well as nitric oxide concentration and erythrocytic lipid peroxidation in lithiatic group. However, MUSA treatment significantly restored the impairment in above kidney function test as that of standard treatment, cystone in a dose-dependent manner. CONCLUSIONS: The present findings demonstrate the efficacy of MUSA in EG-induced urolithiasis, which might be mediated through inhibiting various pathways involved in renal calcium oxalate formation, antioxidant effect, and potential to inhibit biochemical markers of renal impairment.

Alteration in Oxidative/nitrosative imbalance, histochemical expression of osteopontin and antiurolithiatic efficacy of Xanthium strumarium (L.) in ethylene glycol induced urolithiasis.

Xanthium strumarium has traditionally been used in the treatment of urolitiasis especially by the rural people in India, but its antiurolithiatic efficacy was not explored scientifically till now. Therefore, the present study was designed to validate the ethnic practice scientifically, and explore the possible antiurolithiatic effect to rationalize its medicinal use. Urolitiasis was induced in hyperoxaluric rat model by giving 0.75% ethylene glycol (EG) for 28days along with 1% ammonium chloride (AC) for first 14days. Antiurolithiatic effect of aqueous-ethanol extract of Xanthium strumarium bur (xanthium) was evaluated based on urine and serum biochemistry, oxidative/nitrosative stress indices, histopathology, kidney calcium and calcium oxalate content and immunohistochemical expression of matrix glycoprotein, osteopontin (OPN). Administration of EG and AC resulted in hyperoxaluria, crystalluria, hypocalciuria, polyurea, raised serum urea, creatinine, erythrocytic lipid peroxidise and nitric oxide, kidney calcium content as well as crystal deposition in kidney section in lithiatic group rats. However, xanthium treatment significantly restored the impairment in above kidney function test as that of standard treatment, cystone. The up-regulation of OPN was also significantly decreased after xanthium treatment. The present findings demonstrate the curative efficacy of xanthium in ethylene glycol induced urolithiasis, possibly mediated through inhibition of various pathways involved in renal calcium oxalate formation, antioxidant property and down regulation of matrix glycoprotein, OPN. Therefore, future studies may be established to evaluate its efficacy and safety for clinical use.

Toxic effect of sodium fluoride on hydroxyproline level and expression of collagen-1 gene in rat bone and its amelioration by Tamrindus indica L. fruit pulp extract.

Excessive fluoride intoxication plays an important role in the development of dental, skeletal and non-skeletal fluorosis. The aim of this study was to ascertain the toxic effect of excessive fluoride ingestion on the level of hydroxyproline and expression of type 1 collagen gene in rat bone and its amelioration by supplementation with Tamarindus indica fruit pulp extract. Forty albino rats were randomly assigned to four groups. The first group served as control and received only tap water. The second group received sodium fluoride (200 ppm) through drinking water. The third group received T. indica fruit pulp extract (200 mg/kg body weight) alone and the fourth group received the T. indica fruit pulp extract (200 mg/kg body weight) along with fluorinated drinking water (200 ppm) daily by gavage for a period of 90 days. The level of hydroxyproline and expression of type 1 collagen gene using quantitative real time PCR in the tibia bone decreased significantly with continuous exposure to sodium fluoride. Co-administration of T. indica fruit pulp extract during exposure to fluoride through drinking water restored the level of calcium, phosphorus and alkaline phosphatase in serum and the concentration of hydroxyproline in urine. It increased the level of hydroxyproline and expression of type 1 collagen gene in the tibia as compared to untreated fluoride-exposed rats. It is concluded that T. indica fruit pulp extract has an ameliorative potential to protect the bone from fluoride induced collagen damage.

Trypanosoma evansi infection in a German shepherd dog - Apparent successful treatment using serial low dose of diminazene aceturate.

The present study reports a new case of Trypanosoma evansi infection in a dog possibly due to oral transmission from a wild antelope, Nilgai. A four year old male German shepherd dog presented to the Referral Veterinary Polyclinic with history of inappetance, persistent fever, ocular discharge, standing for prolong periods and head pressing. Physical examination revealed corneal opacity, absence of menace reflex and partial blindness. The blood smear examination revealed the presence of Trypanosoma species. T. evansi infection was confirmed by RoTat 1.2 T. evansi specific Polymerase Chain Reaction (PCR). Haemato-biochemical examination showed anaemia, leukopenia, neutrophilia, a mild increase in concentration of alanine aminotransferase, blood urea nitrogen, and creatinine, and a decrease in concentration of blood glucose level. The dog was treated with diminazene aceturate at 3.5mg/kg body weight by deep intramuscular for 5days along with supportive therapy. Marked recovery in clinical signs as well as restoration of normal organ function and oxidant/antioxidant equilibrium was observed after two weeks of treatment. The dog was followed up to 6month and was negative for T. evansi by microscopy and PCR. The present treatment of a consecutive five dose regime of diminazene aceturate along with supportive therapy may help clinicians to overcome the hurdle of relapsing parasitaemia due to T. evansi and successful recovery in clinical cases.

Neurological trypanosomiasis in quinapyramine sulfate-treated horses--a breach of the blood-brain barrier?

Trypanosoma evansi infection typically produces wasting disease, but it can also develop into a neurological or meningoencephalitis form in equids. Trypanosomiasis in horses was treated with quinapyramine sulfate, and all the 14 infected animals were recovered clinically. After clinical recovery, four animals developed a neurological form of the disease at various intervals. Two of these animals treated with diminazene aceturate recovered temporarily. Repeated attempts failed to find the parasite in the blood or the cerebrospinal fluid (CSF), but all of the animals were positive in enzyme-linked immunosorbent assay. The calculation of the antibody index (AI) in the serum and the CSF and polymerase chain reaction (PCR) analysis of the CSF and brain tissue were carried out to confirm the neuro-infection. We found PCR and AI analyses of the CSF to be useful tools in the diagnosis of the neurological form of trypanosomiasis when the organism cannot be found in the blood or CSF. The increased albumin quotient is indicative of barrier leakage due to neuroinflammation. The biochemical changes in the CSF due to nervous system trypanosomiasis include increases in the albumin quotient, total protein, and urea nitrogen. It seems to be the first report on relapse of the nervous form of trypanosomiasis in equids even after quinapyramine treatment in endemic areas.

Hyperlipidaemia in trypanosomiasis of naturally infected horses: possible cachexia-anorexia syndrome?

Trypanosomiasis caused by Trypanosoma evansi commonly produces wasting disease with signs of emaciation and cachexia mainly at the end stage. The present study was conducted to explore the possible hyperlipaemia or hyperlipidaemia and its association with cachexia-anorexia in equine trypanosomiasis. Out of the fifteen confirmed animals, none of the plasma sample was opaque. There was a significant increase in plasma triglyceride, total cholesterol and blood urea nitrogen and a highly significant increase in low-density lipoprotein (LDL) levels. A mild increase in high-density lipoprotein (HDL) and very low-density lipoprotein levels were observed, while the relative percentage of HDL and LDL was altered with high significance. A moderate increase in triglyceride and highly significant increase in LDL might be the reasons for retention of appetite and lipolysis. Possible protein breakdown and presence of lipolysis might be the reasons for cachexia in equine trypanosomiasis.

Correlations among Anaplasma marginale parasitemia and markers of oxidative stress in crossbred calves.

The present study was designed to determine the correlations among Anaplasma marginale parasitemia and markers of oxidative stress in crossbred calves. Blood was collected from 11 crossbred calves infected with A. marginale along with 11 healthy crossbred calves as controls for determination of hematology and oxidative stress indicators. Percentage of parasitemia in infected calves varied from 0.8% to 6.0%. The values of hematological indicators and antioxidant enzymes were decreased, whereas erythrocytic lipid peroxidation (LPO) and plasma nitrate (NO) level were significantly (p < 0.05) augmented in A. marginale-infected animals over healthy group. Parasitemia was positively correlated (p < 0.01) with erythrocytic LPO and plasma NO and negatively correlated (p < 0.01) with hematological indicators and antioxidant enzymes. In addition, erythrocytic LPO was negatively correlated (p < 0.01) with the hemoglobin, erythrocyte count, and packed cell volume. From the present study, it can be concluded that anaplasmosis in crossbred calves is associated with a parasitic load-dependent oxidative damage as indicated by poor antioxidant status and enhanced oxidative stress, which are contributed to severe anemia.

Medicinal herbs: a potential source of toxic metal exposure for man and animals in India.

To explore the possibility of translocation of heavy metals into humans and animals, the authors studied 28 commonly used medicinal plants and estimated their heavy metal content. The plant materials were collected from the same sources used by traditional healers and commercial drug manufacturers. The plants were identified, authenticated, and processed for the analysis of toxic metals. Lead and cadmium levels were estimated in leaf, stem bark, root, or seeds, depending on the medicinal value of the plant portion. The authors used an atomic absorption spectrophotometer to determine levels of metals. The mean lead concentration in medical herbs ranged between 2.624 ppm (standard deviation = 0.426) and 32.757 (standard deviation = 0.124 ppm), and the cadmium concentration ranged between 0.056 ppm (standard deviation = 0.002) and 0.419 ppm (standard deviation = 0.006). Interestingly, the heavy metal concentrations (i.e., lead and cadmium) were higher in leaf than in stem bark or roots, and the lowest values were recorded in seeds. No published reports on the permissible level of toxic metals in commonly used medicinal plants of India have come to the authors' attention; therefore, it was difficult for the authors to determine the role of toxic metals in drug-induced health hazards. However, the presence of toxic metals in different plants led to the conclusion that prolonged consumption of such medicinal plants may be detrimental to health.