Abstract The quality and quantity of the extracted DNA are two key aspects for a successful PCR (Polymerase Chain Reaction) amplification. Moreover, reduction in time and cost required for DNA extraction are also two considerable factors, in cases when large number of samples are to be analyzed within a limited time-frame and budget. Accordingly, the aim of this study was to compare and optimize performance of five different DNA extraction methods by boiling meat tissues from cattle, buffalo, sheep, goat, chicken, camel, horse and dog in PBS (Phosphate Buffer Saline), distilled water, alkaline lysis buffers 1, 2 or 3. The results indicated that the boiling of meat and its products in alkaline lysis buffers was a good method to extract crude DNA. The optimized crude DNA extraction protocol was coupled with PCR-RFLP (Restriction Fragment Length Polymorphism) analysis for meat species differentiation. This developed workflow was tested on fifty-three commercial beef and mutton samples, out of which three samples were found to be adulterated. In conclusion, the rapid crude DNA extraction protocol has led to the development of a direct PCR-RFLP workflow that is simple, time-saving and cost-effective for PCR-based identification of different meat species.

ABSTRACT Huanglongbing (HLB) is one of the world’s worst diseases of citrus trees. The research was conducted to characterize positive citrus samples of two local varieties, ‘Kinnow’ mandarin (Citrus reticulata Blanco) and ‘Mosambi’ orange (C. sinensis (L.) Osbeck), based on morphological, physical and biochemical characters. Physical and biochemical parameters of greening infected and healthy samples of both the cultivars were significantly different. Citrus fruits were lopsided and half to full green in the HLB infected trees compared to healthy. The HLB infected leaves samples were small and narrow. Fruit diameter, leaf area, and juice weight of HLB infected samples were significantly less than healthy samples. The rind thickness of infected fruits (70% and 80%) was significantly higher than healthy ones (20% and 30%). Starch content of leaves from mandarin (5.54 μg mL-1) and orange (6.12 μg mL-1) were significantly higher in HLB infected plants, while juice percentage of HLB infected fruit samples was lesser in mandarin and orange (22.73%, and 14.70%, respectively) than healthy fruits (38.22% and 31.30%, respectively). Acidity was significantly increased in juice of HLB infected mandarin and orange fruits (0.57%, and 0.53%, respectively). Similarly, there was significant reduction in biochemical parameters; total sugars in mandarin and orange (5.16% and 4.79%), total soluble solids (8.54% and 8.28%), total chlorophyll (13.50 and 14.92 mg mL-1) and ascorbic acid (34.28% and 34.12%) in HLB infected samples of both cultivars. This study determined differences of physical and biochemical parameters of two local citrus varieties, commonly cultivated in the Asian region.

Abstract Fenugreek leaves contains bioactive compounds, which are helpful in maintaining human health. These compounds are adversely affected in conventional drying methods. Therefore, this study was carried out to retain maximum amount of these components through microwave-assisted drying and extraction technique. The technique was optimized for simultaneous drying and extraction process for fenugreek leaves. The drying rate and drying time were influenced by the level of microwave power; time was reduced from 21 to 5 minutes, when microwave power was changed from 30 to 100 W. Unlikely, the drying rate increased from 1.79 to 4.56 ± 0.05 g/(100 g.min) with the change in power (30 to 100 W). Moreover, moisture ratio analysis determined that two term model was the best to demonstrate the fitness with experimental values. The energy consumption was lowest for 100W compared to other powers. Furthermore, comparison between microwave-based extraction and methanol-extraction indicated that the antioxidant activity was better preserved compared to conventional ones even at higher microwave power levels.

Abstract In this study, the physicochemical properties of walnut (Juglans regia L.) oils collected from different sites in Khyber PakhtoonKhwa (KPK), Pakistan were investigated. The obtained walnut oils showed variation in viscosity, surface tension, density, acid, saponification, and iodine values. The oil obtained from the Madyan (Sample E) showed the lowest density (0.981 g/cm3), surface tension (34.76×10-3 N/m), viscosity (15.0595 cp) saponification values 84.15 (mg KOH/g of oil) and acid value 3.5 (mg KOH per gram oil) amongst all. The physicochemical properties of all the oils were also compared to olive oil (standard) where sample E showed similarities in such properties compared to olive oil. Moreover, higher concentrations of oleic acid (41 %) and linoleic acid (9 %) were detected in the walnut oil of sample E in GC/MS analysis. The similarities of walnut oil obtained from Madyan to that of standard (olive oil) thus affirm it as best quality oil and suitable for food processing purposes.

Resumen En este estudio se investigaron las propiedades fisicoquímicas del aceite de nuez (Juglans regia L.) recolectado de diferentes sitios en Khyber PakhtoonKhwa (KPK), Pakistán. Los aceites de nuez obtenidos mostraron variación en los valores de viscosidad, tensión superficial, densidad, acidez, saponificación e índice de yodo. El aceite obtenido del Madyan (Muestra E) mostró la menor densidad (0.981 g/cm3), tensión superficial (34.76 × 10-3 N/m), viscosidad (15.0595 cp), valores de saponificación 84.15 (mg KOH/g de aceite) y valor ácido 3.5 (mg KOH por gramo de aceite) entre otros. Las propiedades fisicoquímicas de todos los aceites también se compararon con el aceite de oliva (estándar); la muestra E mostró similitudes en las diferentes propiedades en comparación con el aceite de oliva. Además, se determinaron concentraciones más elevadas de ácido oleico (41 %) y ácido linoleico (9 %) en el aceite de nuez de la muestra E mediante el análisis GC/MS. Las similitudes del aceite de nuez obtenido de Madyan con el estándar (aceite de oliva) lo afirman como aceite de mejor calidad y adecuado para el procesamiento de alimentos.

ABSTRACT GeneXpert is one of the recent technological instruments used to diagnose tuberculosis in a short span of time. In this study, the performance of GeneXpert MTB/RIF assay for the diagnosis of extrapulmonary tuberculosis (EPTB) was compared with light-emitting diode Fluorescent Microscopy (LED-FM) in Khyber Pakhtunkhwa, Pakistan. A total of 737 EPTB samples were collected from tuberculosis (TB) suspected patients. Out of these samples, male to female ratio was 53% (n = 390) to 47% (n = 347) respectively. The sensitivity and specificity was 73% and 100% for GeneXpert, while 40% and 100% for LED-FM microscopy. This shows that the sensitivity of GeneXpert is 40–50%, higher than LED-FM microscopy. GeneXpert also detected low number of bacilli as compared to LED-FM microscopy.

Abstract In this study for the first-time microbial communities in the caves located in the mountain range of Hindu Kush were evaluated. The samples were analyzed using culture-independent (16S rRNA gene amplicon sequencing) and culture-dependent methods. The amplicon sequencing results revealed a broad taxonomic diversity, including 21 phyla and 20 candidate phyla. Proteobacteria were dominant in both caves, followed by Bacteroidetes, Actinobacteria, Firmicutes, Verrucomicrobia, Planctomycetes, and the archaeal phylum Euryarchaeota. Representative operational taxonomic units from Koat Maqbari Ghaar and Smasse-Rawo Ghaar were grouped into 235 and 445 different genera, respectively. Comparative analysis of the cultured bacterial isolates revealed distinct bacterial taxonomic profiles in the studied caves dominated by Proteobacteria in Koat Maqbari Ghaar and Firmicutes in Smasse-Rawo Ghaar. Majority of those isolates were associated with the genera Pseudomonas and Bacillus. Thirty strains among the identified isolates from both caves showed antimicrobial activity. Overall, the present study gave insight into the great bacterial taxonomic diversity and antimicrobial potential of the isolates from the previously uncharacterized caves located in the world's highest mountains range in the Indian sub-continent.