Beautiful green leaves of Papaya are the rich source of Chlorophyll. Green color of chlorophyll has been used for a very long time as a natural colorant. Carica papaya has been considered as a good example and reasonable source of natural phytochemicals, which makes it suitable to color the food items and beverages. The aim of the present investigation is to develop the process of ultrasonic extraction in combination with solid phase extraction (SPE) to extract out chlorophyll with high yield as well as high degree of clarity. Newly customized ultrasonic-assisted extraction technique for the extraction of chlorophyll from Carica papaya leaves is optimized by taking different parameters like time, temperature, solvents concentrations, and raw material under consideration. Furthermore, the extract was purified by means of SPE and examined by using UV-Vis spectrophotometer. The highest yield of chlorophyll (dye) extract was found as 40% in solvent solution having 80 mL of ethanol and 20 mL of water with 5 minutes of extraction time, 35°C of temperature, and 1 grams of raw material in the sonication bath. Furthermore, the SPE purified sample was characterized by means of the UV-Vis spectrophotometer and here the total chlorophyll content was 34 mg/g, including chlorophyll a with a concentration of 14.1246 mg/g and chlorophyll b with concentration of 19.845 mg/g respectively. Consequently, sonication method can be suggested as a good method to get better concentration of chlorophyll.

Feather wastes-byproduct of commercial poultry processing plant is produced in large amounts. Keratinolytic enzymes produced by feather degrading bacteria can easily degrade these waste products releasing pure keratin as a residue. The aim of present study was to isolate, and characterize feather degrading bacteria as well as assess the keratinolytic potential of purified enzyme. Three feather degrading bacteria (dps3, wps1 and dcs1) were isolated from feathers of domestic chickens. Preliminary characterization of isolated bacteria revealed these isolates belonging to genus Bacillus. 16S rRNA gene sequencing identified the isolates as B. subtilis dps3 (MW255302), B. cereus wps1 (MW255303) and B. licheniformis dcs1 (MW255304). Cell free supernatant of B. licheniformis dcs1 degraded feathers completely in 14 days indicating its keratinolytic ability. Purification of keratinase enzyme from B. licheniformis dcs1 was performed using column chromatography. SDS-PAGE indicated its molecular weight as 32 KDa. Kerotinolytice activity was maximum at optimum pH of 7 and 45°C temperature. Enzyme showed the potential to degrade keratin material such as hairs and nails of humans. Findings of current study suggested that purified enzyme possess potential to upgrade nutritional quality of poultry waste containing keratin and might play as important biotechnological tool for keratin hydrolysis.

The extremely difficult and challenging process is identifying pheretimoid species, genus Metaphire and Amynthas involving increased homoplasy in various morphological characteristics. The molecular identification, phylogenetic relationships, and evolutionary divergence time of earthworms belonging to the pheretimoid complex were investigated in this study using partial mitochondrial COI (cytochrome C oxidase subunit I) gene sequences ranging from 550-680 bp. Results revealed that 86 pheretimoid earthworms were morphologically different from a total of 342 mature worms. Moreover, 11 pheretimoid species were molecularly identified, including Metaphire posthuma (02), M. anomala (01), M. houlleti (02), M. californica (01), M. birmanica (02), Amynthas minimus (01), A. morrisi (01), and M. bununa (01). A phylogenetic tree was constructed with bootstrap values of 95%, which supported a monophyletic lineage of two well-supported clades formed by 12 partial COI sequences and 48 GenBank sequences using Hirudo medicinalis as an outgroup. The monophyly of these obtained genera indicated overall similarity at species level. Today, species like Amynthas, Metaphire and Pheretima have worm diversity in the form of pheretimoid earthworms, which dates to the Late Miocene (11.2-5.3 Mya) and the Pliocene (5.3-2.4 Mya). Compared to all relevant pheretimoid species, genetic p-distance values ranged from 0.0% to 0.57% (less than 1%). These low range values demonstrated that both genera Metaphire and Amynthas, supported the theory, which states that there are shared similarities among the species, despite different morphology. The current study is the first attempt in Pakistan to identify earthworms through DNA barcoding thus providing a genomic stamp. The work explored the significance of COI gene sequences to construct molecular tools that will be useful to overcome the different obstacles in morphologically similar earthworm identification and their phylogenetic study.

In developing countries, the chances of fraud in written documents are comparatively high. Therefore, comparison of fountain pen inks is especially imperative in examination of forensic questioned documents. We have investigated the use of the gas chromatography-mass spectrometry technique in profiling and discrimination of fountain pen ink used in Pakistan for forensic purpose. The main purpose of this study was to discriminate different Pakistani fountain pen inks. The datum for Pakistani inks of fountain pen is not obtainable. In this research study, blue, black, and green colors fountain pen inks commercially used in Pakistan have been extracted from paper using micropunch and then investigated using the gas chromatography-mass spectrometry technique. Gas chromatography-mass spectrometry (GC-MS) was used to differentiate various brands of different colors of fountain pen inks based on their chemical composition. Molecular ion peaks for different components were obtained, and components were identified on the basis of detected ions. Results have been calculated and compared in terms of discriminating power (D.P.). The D.P. for blue, black, and green inks of fountain pen was 1.0 by using the gas chromatography-mass spectrometry technique.

Calcium is a dynamic mineral. Recent discoveries designate that low intake of calcium generates deficiencies and path to other diseases. Food fortification could play a key role to overcome this problem. To cope with this deficiency problem, jellies were formulated with food-grade calcium salts and chicken eggshell powder. In the present study, three different concentrations of calcium salts, as well as eggshell powder were used to formulate jellies. The results of the sensory evaluation indicated that the two jelly products (A&D) in the current study were suitable for consumers. Results of Atomic Absorption Spectrophotometer revealed Jelly A and jelly D had 151±0.05 ppm and 133±0.06 ppm calcium concentration, respectively. Proximate analysis of Jelly A showed that it has 6.0±0.01% ash, 9.2±0.1% moisture, 0.4±0.01 g crude protein, 82.79±0.001 g crude fiber, and 0.61±0.001 g crude fat, while the jelly D that was made with chicken eggshell powder exhibited 6.0±0.01% ash, 10.1±0.1% moisture, 0.5±0.01 g protein, 84.54±0.01 g crude fiber and 1.61±0.01 g crude fat. Therefore, these two jelly A & D were greatly appreciated among other attributes. In spite of naturally available calcium-rich sources, calcium-fortified jellies can be consumed by individuals who are incapable to take sufficient calcium from their diet.