Sanjeev Kumar Shukla

High-performance liquid chromatography method validation for determination of tetracycline residues in poultry meat

Background: In this study, a method for determination of tetracycline (TC) residues in poultry with the help of high-performance liquid chromatography technique was validated. Materials and Methods: The principle step involved in ultrasonic-assisted extraction of TCs from poultry samples by 2 ml of 20% trichloroacetic acid and phosphate buffer (pH 4), which gave a clearer supernatant and high recovery, followed by centrifugation and purification by using 0.22 μm filter paper. Results: Validity study of the method revealed that all obtained calibration curves showed good linearity ( r 2 > 0.999) over the range of 40-4500 ng. Sensitivity was found to be 1.54 and 1.80 ng for oxytetracycline (OTC) and TC. Accuracy was in the range of 87.94-96.20% and 72.40-79.84% for meat. Precision was lower than 10% in all cases indicating that the method can be used as a validated method. Limit of detection was found to be 4.8 and 5.10 ng for OTC and TC, respectively. The corresponding values of limit of quantitation were 11 and 12 ng. Conclusion: The method reliably identifies and quantifies the selected TC and OTC in the reconstituted poultry meat in the low and sub-nanogram range and can be applied in any laboratory.

Comparison of antimicrobial efficacy of a fixed dose combination of ceftazidime + sulbactam with ceftazidime and sulbactam alone against five bacteria

The efficacy of the agents was evaluated on the basis of minimum inhibitory concentration (MIC) and time-kill curve analysis in Pseudomonas aeruginosa, Staphylococcus aureus, Acinetobacter baumannii, Mycobacterium smegmatis, and Citrobacter braakii; MIC was found to be 0.5, 4.0, 0.015625, 0.0078125, and 0.0625 mg/L in FDC of ceftazidime + sulbactam, respectively, which is lower than ceftazidime and sulbactam individually. Time-kill curve analysis demonstrated maximum killing of bacteria after 4 h. A fixed dose combination of ceftazidime + sulbactam was found to have stronger antibacterial properties than ceftazidime and sulbactam alone at in vitro analysis.

Molecular Characterization of Mitochondrial D Loop of Indian Red Jungle Fowl

Mitochondrial DNA has the characteristic of quick evolution, matrilineal inheritance, and simple molecular structure, and it serves as the most used marker for molecular study. As an important role of genomics, studying it can help understand the origins, history, and adaptation of domestication. A 1325 bp fragment of the mtDNA D-loop region was amplified, sequenced and phylogenetic study was done. Out of 400 sites about 12 % were polymorphic between the Gallus gallus (G.g.) subspecies and most of them (60%) were transitions. Indian RJF showed very low genetic distances (0.008-0.013) with G.g. gallus birds from Thailand as compared to those from Japan and Indonesia. The G.g. murghi showed comparatively high genetic distance i.e. 031-0.034 with G.g. spadiceus and from 0.061-0.066 with G. gallus bankiva. The phylogenetic tree showed that G.g. banikva is well separated from the other three subspecies as it made a separate cluster. G.g. gallus makes two separate clusters i.e. one those from Thailand and other those from Japan and Indonesia. Within each chicken breed, there is an excess of homozygosity, but there is no significant reduction in the nucleotide diversity. Phenotypic modifications of chicken breeds as a result of artificial selection appear to stem from ancestral polymorphisms at a limited number of genetic loci.

Assessment of Immunocompetence Status of Native Breed of Chickens

Immunocompetence is known to be controlled by genetics depending on the B-haplotype Major Histocompatibility complex (MHC), for instance and the selection for production traits. Genetic resistance to disease involves many facets of the body’s defense system and interactions, and it is extremely complex. Production traits had negative correlation to disease resistant traits. The aim of the current study was to estimate the immune responses against SRBC in selected lines, estimating the genetic trend and genetic correlation between male and female immune responses against SRBC. Humoral response was estimated in the form of titer against sheep RBC, each birds was received an intravenous injection of 0.1 ml of 0.5% suspension of packed SRBC in normal saline at 6 week of age Five day later the blood was collected from Jugular vein of each birds in the individual test tube and sera were collected. In the present investigation, chicks belonging to the selected lines of native chickens were used for the study of immunological traits viz., response to sheep RBCs, IgG concentrations. Finally, 1% sheep RBC suspension was prepared by mixing 1 ml of packed sheep RBCs and 99 ml of PBS which was then used for injection in the birds. The Average means for HA was observed (7.74±0.109) and IgG (1.94±0.72) in native breed. Sex wise HA results showed that male have higher mean (7.925 ± 0.1463) than females (7.57± 0.161) for HA while the same was also estimated for IgG by having mean (2.1194±8.9) and female (1.77±8.18). Test of Homogeneity of Variances was recorded as (0.39) for HA which was significant and IgG level (0.937).