Yellow scorpion (Buthus sinidicus) venom peptides induce mitochondrial-mediated apoptosis in cervical, prostate and brain tumor cell lines.

Scorpion venoms are known to contain over 100,000 biologically active constituents. However, only a few of them have been studied. The major constituents of venom are proteins and peptides, which exhibit various biological and pharmacological properties, including anticancer activities. In the current study, the venom of yellow scorpions (Buthus sindicus) found in Sindh, Pakistan, was extracted and evaluated for its anti-cancer and anti-inflammatory activities. The crude venom showed a dose dependent inhibition of phagocyte oxidative burst from human whole blood cells (28.3% inhibition at highest tested concentration of 300 µg/mL). In-vitro cytotoxicity of crude venom was evaluated against human prostrate (PC3), cervical (HeLa) and neuroblastoma (U87-MG) cell lines, along with cytotoxicity against normal human fibroblast (BJ) cells. Crude venom was cytotoxic to all cell lines, with prominent inhibitory effect on PC3 cells. Crude venom was fractionated through RP-UPLC, resulted in fifteen fractions, followed by evaluation of their anticancer potential. Among all, the fraction I significantly (P < 0.001) reduced the cell viability of all three cancer cell lines, and exhibited insignificant cytotoxicity against normal cell line. Furthermore, the apoptotic cell death pathway was evaluated for crude venom, and fraction I, in most sensitive cell line PC3, by using flow-cytometry analysis. Both crude venom and its fraction I caused a mitochondrial-mediated apoptosis in prostate cancer cells (PC3). To the best of our knowledge, this is the first report of the anticancer and anti-inflammatory activity of venom of Pakistani yellow scorpions. Results indicate their therapeutic potential, and a case for further purification and validation studies.

Nephroprotective effect of ethanol extract and fractions of a sea lettuce, Ulva fasciata against cisplatin-induced kidney injury in rats.

Cisplatin is widely used in anticancer therapy, but a substantial percentage of patients who receive the therapeutic dose of cisplatin develop nephrotoxicity. Hepatotoxicity may also develop after a single dose or low repeated doses of cisplatin. Ulva fasciata is an edible seaweed, commonly known as sea lettuces have also been shown various biological activities. In this study, ethanol extract and its solvent fractions (n-hexane and chloroform) of U. fasciata were given (orally) to different groups of rats for 10 days. Injury to the kidney was induced by administrating cisplatin, intraperitoneally (i.p.) to rats at a dose of 7 mg/kg body weight (b.w.) dissolved in 1 mL saline, at 5th day of the experiment. At 10th day rats were sacrificed and kidney parameters (creatinine, urea, and blood urea nitrogen (BUN)) and electrolyte balance (Ca++, Mg++, K+, and Na+) in serum were determined, while oxidative stress markers glutathione (GSH), catalase (CAT) and malondialdehyde (MDA), and inflammatory cytokines, tumor necrosis factor (TNF α), and interleukin (IL-6) were determined in kidney tissues. Histological examination of the kidney was also performed to examine the changes in kidney tissues. Cisplatin caused adverse effects on blood parameters, antioxidants, and inflammatory markers with severe renal tubular injury in kidney tissues. Ethanol extract of U. fasciata and its fractions effectively improved these disorders and diminished the renal dysfunction. However, ethanol extract was found more effective in attenuating the adverse effect of cisplatin than its fractions. n-Hexane-soluble fraction that was subjected to GC-FID and GC-MS analysis revealed the presence of several compounds and some of them are new from this source. It could be concluded that the U. fasciata possesses nephroprotective effect and can attenuate cisplatin-induced renal dysfunction. Since U. fasciata is an edible seaweed, it may be used as a diet supplement.

Informing water distribution line rehabilitation through quantitative microbial risk assessment.

Poor urban water quality has been linked to diminished source water quality, poorly functioning water treatment systems and infiltration into distribution lines after treatment resulting in microbiological contamination. With limited funding to rehabilitate distribution lines, developing nations need tools to identify the areas of greatest concern to human health so as to target cost effective remediation approaches. Herein, a case study of Hyderabad, Pakistan was used to demonstrate the efficacy of combining quantitative microbial risk assessment (QMRA) for multiple pathogens with spatial distribution system modeling to identify areas for pipe rehabilitation. Abundance of Escherichia coli, Enterococcus (enterococci), Salmonella spp., Shigella spp., Giardia intestinalis, Vibrio cholera, norovirus GI and adenovirus 40/41, were determined in 85 locations including the source water, treatment plant effluent and the city distribution lines. Bayesian statistics and Monte Carlo simulations were used in the QMRA to account for left-censored microbial abundance distributions. Bacterial and viral abundances in the distribution system samples decreased as follows: 9400 ± 19,800 norovirus gene copies/100 mL (average ± standard deviation, 100% of samples positive); 340 ± 2200 enterococci CFU/100 mL (94%), 71 ± 97 Shigella sp. CFU/100 mL (97%), 60 ± 360 E. coli CFU/100 mL (89%), 35 ± 79 adenovirus gene copies/100 mL (100%), and 21 ± 46 Salmonella sp. CFU/100 mL (76%). The QMRA revealed unacceptable probabilities of illness (>1 in 10,000 illness level) from the four exposure routes considered (drinking water, or only showering, tooth brushing, and rinsing vegetables consumed raw). Disease severity indices based on the QMRA combined with mapping the distribution system revealed areas for targeted rehabilitation. The combined intensive sampling, risk assessment and mapping can be used in low- and middle-income countries to target distribution system rehabilitation efforts and improve health outcomes.

Metagenomic analysis of drinking water samples collected from treatment plants of Hyderabad City and Mehran University Employees Cooperative Housing Society.

The quality assessment of water, supplied to the end user, is an essential part to assess the physical, chemical, and biological status of water, which impacts on human health. For the quality assessment of drinking water treatment plants and distribution systems of Hyderabad City and Mehran University of Engineering and Technology, Jamshoro, Pakistan, 13 surface drinking water samples were collected from three treatment plants, two of Hyderabad City, including WASA treatment plant and its distribution system (n = 5), Hala Nakka treatment plant and its distribution system (n = 6), and Mehran University Employees Cooperative Housing Society (MUECHS) treatment plant and its distribution system (n = 2). Physicochemical parameters of all drinking water samples were in the range compared to EPA and WHO guidelines, except in L-12 sample. Notably, no free-chlorine was detected in all samples. In metagenomics analysis, targeting V3-V4 hypervariable region of 16S rRNA gene, in QIIME2 environment, high bacterial prevalence was observed in all samples. On average, 348 OTUs were observed per sample. Among all samples, treated water sample from the Hala Nakka Treatment Plant (HNTR) was the most diverse sample in bacterial composition (Shannon 7.51 and Simpsons reciprocal indices 0.98). Overall, Proteobacteria, Bacteroidetes, Cyanobacteria, Verrucomicrobia, and Actinobacteria were the five most abundant phyla (relative abundances of 43.6, 37.9, 8.5, 2.5, and 2.4 percent, respectively). Notably, Cyanobacteria are well-known toxin producers which effect the human, and animal health. At genus level, Flavobacterium (4.86%) and Aquirestis (3.77%) were the most abundant genera. Functional predictions, based on 16S rRNA gene by PICRUSt, predicted 6909 KEGG orthologies, relating to 245 KEGG pathways. Among the predicted pathways of KEGG orthologies, pathways to human infections were also found. In conclusion, this study gave a deep insight into bacterial contamination in drinking water samples of Hyderabad City and MUECHS treatment plants and water quality status in Hyderabad and Mehran University of Engineering and Technology.

Metagenomic Characterization of Bacterial Communities in Drinking Water Supply System of a Mega City.

Supplying safe water to consumers is vital for protection of public health. With population of > 15 million, Karachi is the main economical hub of Pakistan. Lake Keenjhar serves as the main source of fresh water while Hub dam is the secondary water reservoir for Karachi. In this study, bacterial community of the drinking water supply system (DWSS) of Karachi was studied from source to tap using metagenomics approach. For this purpose, we collected 41 water samples from different areas of the city (n = 38) and water reservoirs (n = 3). 16S rDNA metagenomic sequencing of water samples revealed that 88% sequences were associated with Proteobacteria (52%), Planctomycetes (15%), Becteroidetes (12%), and Verrucomicrobia (6%). On the class level, α-proteobacteria (6-56%) were found to be the most abundant followed by ß- (8-41%) and γ-proteobacteria (6-52%). On the genus level, substantial diversity was observed among the samples. Bacterial communities in water from Hub dam was found to be distantly related while among the residential towns, Lyari was highly distant from the others. Twenty-four bacterial genera were found to be exclusively present in residential area samples in comparison to the source waters which is suggestive of their resistance against treatment procedures and/or contamination. Metagenomic analysis revealed abundance of Pseudomonas, Legionella, Neisseria, Acinetobacter, Bosea, and Microcystis genera in residential areas water samples. The present metagenomic analysis of DWSS of Karachi has allowed the evaluation of bacterial communities in source water and the water being supplied to the city. Moreover, measurement of heavy metals in water samples from Karachi revealed arsenic concentration according to WHO standards which is in contrast of recent study which reported extensive arsenic contamination in aquifers in the Indus valley plain. To the best of our knowledge, this is the first metagenomic study of DWSS of Karachi.

Characterization of gut bacterial flora of Apis mellifera from north-west Pakistan.

Gut microbiota has been recognized to play a beneficial role in honey bees (Apis mellifera). Present study was designed to characterize the gut bacterial flora of honey bees in north-west Pakistan. Total 150 aerobic and facultative anaerobic bacteria from guts of 45 worker bees were characterized using biochemical assays and 16S rDNA sequencing followed by bioinformatics analysis. The gut isolates were classified into three bacterial phyla of Firmicutes (60%), Proteobacteria (26%) and Actinobacteria (14%). Most of the isolates belonged to genera and families of Staphylococcus, Bacillus, Enterococcus, Ochrobactrum, Sphingomonas, Ralstonia, Enterobacteriaceae, Corynebacterium and Micrococcineae. Many of these bacteria were tolerant to acidic environments and fermented sugars, hence considered beneficial gut inhabitants and involved the maintenance of a healthy microbiota. However, several opportunistic commensals that proliferate in the hive environment including members Staphylococcus haemolyticus group and Sphingomonas paucimobilis were also identified. This is the first report on bee gut microbiota from north-west Pakistan geographically situated at the crossroads of Indian subcontinent and central Asia.