Amit Raj Gupta

Haematology and Serum Biochemistry of Chital (Axis axis) and Barking Deer (Muntiacus muntjak) Reared in Semi-Captivity

Haematological and serum biochemical values of clinical significance that could serve as reference data for deer kept in captivity were measured for chital (Axis axis) and barking deer (Muntiacus muntjak). The venous blood from four each of chital and barking deer (n = 8) reared in semi-captivity was collected after proper restraint of the animals. The mean blood haemoglobin, packed cell volume, total erythrocyte count and total leukocyte count of all the eight deer of the two species were 15.90 ± 0.44,g/dl, 51.44 ± 0.60%, 20.83 ± 0.57 × 106μl and 2.37 ± 0.20 × 103μl. Serum total protein, albumin, bilirubin, cholesterol and blood urea nitrogen irrespective of species were 6.83 ± 0.19,g/dl, 3.90 ± 0.11,g/dl, 0.33 ± 0.08,mg/dl, 106.81 ± 3.59,mg/dl and 24.79 ± 2.11,mg/dl, respectively. Serum enzyme activities indicative of liver function such as alanine transaminase (ALT) and aspartate transaminase (AST) were 30.38 ± 4.67,units/ml and 42.88 ± 5.97,units/ml, respectively. The serum calcium and phosphorus levels of all the eight deer were 10.27 ± 0.36,mg/dl and 8.31 ± 0.68,mg/dl, respectively. This is the first report on baseline values in barking deer. The distribution of haematological and serum biochemical values was fairly normal, suggesting that the mean values could be representative of normal values for two different deer species.

Sequence characterization and polymerase chain reaction-restriction fragment length polymorphism of the mitochondrial DNA 12S rRNA gene provides a method for species identification of Indian deer

Characterization of species-specific molecular markers and development of a method for identification of Indian deer species is necessary to monitor illegal trade of parts and products for better conservation and management of the endangered species. In this investigation, we characterized the 12S rRNA gene sequence for differentiation of Indian deer species and developed a polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP)-based method for their identification. Universal primers were used for the amplification of the mitochondrial 12S rRNA gene from genomic DNA of chital or spotted deer, hog deer, barking deer, sika deer, musk deer and sambar. PCR products of chital, hog deer and Himalayan musk deer were cloned and sequenced for the first time. Among the Indian deer species, more than 90% similarity was observed in the mitochondrial 12S rRNA gene. The sequences of the above deer species were restriction mapped with the help of Lasergene (DNAstar Inc., Madison, WI, USA). PCR amplicon of these deer species were subjected to restriction digestion with Rsa1, Dde1, Bsr1 and BstSF1 endonucleases that showed a species-specific RFLP pattern. This technique provides a reliable and efficient tool for identification of deer species using a variety of biomaterials.

Original Article. 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

Abstract 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.

Down-regulation of expression of type 1 collagen gene in dentin of fluoride exposed rats.

Excessive ingestion of fluoride leads to abnormal mineralization of both enamel and dentin. Mechanisms about enamel fluorosis have been clarified. The knowledge about dentin fluorosis is limited and the mechanism is unclear. The present experiment was aimed to ascertain the excessive ingestion of fluoride on collagen metabolism in dentin of incisor teeth of rats. Rats (n= 12) were divided into two groups. Rats of group I served as control and received only tap water and those of group II received sodium fluoride @ 200 mg/L of drinking water ad lib for a period of 90 days. Quantitative real-time polymerase chain reaction was used to quantify the gene expression levels of Collal in dentin. Significant decreases in expression of Collal gene in the dentin of incisor teeth of fluoride exposed rats was noted at the end of the experiment.