Examining drinking water quality: analysis of physico-chemical properties and bacterial contamination with health implications for Shangla district, Khyber Pakhtunkhwa, Pakistan.

A comprehensive understanding of water quality is essential for assessing the complex relationship between surface water and sources of pollution. Primarily, surface water pollution is linked to human and animal waste discharges. This study aimed to investigate the physico-chemical characteristics of drinking water under both dry and wet conditions, assess the extent of bacterial contamination in samples collected from various locations in District Shangla, and evaluate potential health risks associated with consuming contaminated water within local communities. For this purpose, 120 groundwater and surface water samples were randomly collected from various sources such as storage tanks, user sites, streams, ponds and rivers in the study area. The results revealed that in Bisham, lakes had the highest fecal coliform levels among seven tested sources, followed by protected wells, reservoirs, downstream sources, springs, rivers, and ditches; while in Alpuri, nearly 80% of samples from five sources contained fecal coliform bacteria. Similarly, it was observed that the turbidity level, total dissolved solids, electrical conductivity, biological oxygen demand, and dissolved oxygen in the surface drinking water sources of Bisham were significantly higher than those in the surface drinking water sources of Alpuri. Furthermore, the results showed that in the Alpuri region, 14% of the population suffers from dysentery, 27% from diarrhea, 22% from cholera, 13% from hepatitis A, and 16% and 8% from typhoid and kidney problems, respectively, while in the Bisham area, 24% of residents are affected by diarrhea, 17% by cholera and typhoid, 15% by hepatitis A, 14% by dysentery, and 13% by kidney problems. These findings underscore the urgent need for improved water quality management practices and public health interventions to mitigate the risks associated with contaminated drinking water. It is recommended to implement regular water quality monitoring programs, enhance sanitation infrastructure, and raise awareness among local communities about the importance of safe drinking water practices to safeguard public health.

Bactericidal role of epidermal mucus of freshwater fish treated with Aeromonas hydrophila.

This study explored the bactericidal role of the epidermal mucus (EM) of five freshwater Cyprinid fish species namely Ctenopharyngodon idella, Labeo rohita, Catla catla, Hypophthalmichthys molitrix, and Cirrhinus mrigala after treatment with Aeromonas hydrophila. Extracts of EM (crude and acidic) of each species showed bactericidal activity against various Gram -ve (Pseudomonas aeruginosa, Escherichia coli, Aeromonas hydrophila, Edwardsiella tarda, Salmonella enterica, Klebsiella pneumonia, Serratia marcescens, and Enterobacter cloacae) and Gram +ve (Bacillus wiedmannii and Staphylococcus aureus) bacteria compared with standard antibiotics (Fosfomycin). The zone of inhibition (ZOI) was measured in millimetres against antibiotics (Fosfomycin). Variations in bactericidal activity of EM were observed against bacteria from the same and different fish species. The acidic extract was more effective than the crude extract and showed significantly higher ZOI values against various bacteria and Fosfomycin antibiotics. This result shows that fish EM may perform an important role in fish defence against bacteria. Therefore, this study may hint towards the substitution of synthetic antibiotics with fish EM that may be used as a novel 'bactericidal' in aquaculture as well as in humans against bacterial infections.

Bullying in educational institutions: college students' experiences.

Despite its widespread prevalence, bullying in educational institutions is not considered a serious social issue in Pakistan. Present study explores the understandings and experiences of college students about bullying and documents the status of anti-bullying interventions in educational institutions. This study is based on in-depth interviews with students and principals of private sector colleges in Pakistan. The study showcases the important aspects of bullying that hurt the educational, personal and social lives of students. The most frequent form of bullying experienced by the students constitutes verbal bullying. Bullying occurs at specific places on campuses which are less supervised by teachers and administration. Although the principals were of the view that they had zero tolerance towards bullying in their premises, they did not have any preventive measures. There is a dire need to develop a policy to address the issue of bullying in educational institutions.

Molecular epidemiology of Shigella flexneri isolated from pediatrics in a diarrhea-endemic area of Khyber Pakhtunkhwa, Pakistan.

Shigella flexneri is considered as an important causative agent of Shigellosis causing diarrhea in the countries with a low socioeconomic status. No study has been carried out on the molecular prevalence of S. flexneri in Khyber Pakhtunkhwa, Pakistan. So this study was designed to evaluate the molecular prevalence of S. flexneri and their associated risk factors. A total of 2014 diarrheal stool samples were collected from January 2016 to May 2017 from pediatrics patients of Khyber Pakhtunkhwa followed by identification of S. flexneri through biochemical, serological, and molecular methods. The overall prevalence of Shigella species was found to be 7.9% (n = 160). The predominant Shigella specie was S. flexneri (n = 155, 96.8%) followed by S. boydii (n = 5, 3.1%). Interestingly, no sample was found positive for S. sonnei and S. dysenteriae. The majority of Shigellosis cases occurred from June to September. Potential risk factors related with Shigellosis were unhygienic latrine usage, bad hand washing, and consumption of unhygienic food and water, and pipe leakage in the sewage system. In this study, we have observed a high number of Shigellosis cases especially those caused by S. flexneri. It is suggested that effective health awareness programs should be organized by the regional health authorities to minimize the magnitude of pediatrics Shigellosis.

A New Construction of High Performance LDPC Matrices for Mobile Networks.

Secure and reliable information flow is one of the main challenges in social IoT and mobile networks. Information flow and data integrity is still an open research problem. In this paper, we develop new methods of constructing systematic and regular Low-Density Parity-Check Matrices (LDPCM), inspired by the structure of the Sarrus method and geometric designs. Furthermore, these codes have cyclic structure and therefore, are less complex in computation and also require less memory in hardware implementation. Besides, an optimal method of post-processing for deleting girths four is presented. Numerical results show that the codes constructed by these methods perform well over the additive white Gaussian noise (AWGN) channel when decoded with the sum-product LDPC iterative algorithms. The proposed methods can be very efficient in terms of reducing memory consumption and improving the convergence speed of the decoder particularly in IoT and mobile networks.

Association of sequence variants in frizzled-6 with autosomal recessive nail dysplasia (NDNC-10) in Pashtun families.

Primitive epidermis develops the nail apparatus. Nails have a strong and inflexible nail plate at the end of each digit. Very few genes responsible for causing nonsyndromic form of nail dysplasia have been reported. In the current study, peripheral blood samples were collectedfrom three unaffected individuals and four affectedindividuals of Family A, while blood from two affected and three unaffected individuals were taken of Family B. Genotyping in both the families was performed using highly polymorphic short tandem repeat microsatellite markers. Sanger sequence of the FZD6 gene was performed and analysed for segregation analysis. A comparative modelling approach was used to predict the three-dimensional structures of FZD-6 protein using Modeller 4. Linkage analysis mapped a disease locus on chromosome 8q22.3, harbouring FZD6. Targeted Sanger sequencing of all the coding exons of FZD6 revealed a nonsense sequence variant in pedigree A, whereas a missense sequence variant in pedigree B. Finding and literature indicates the disease spectrum of Pakistani population with claw-shaped nail dysplasia, particularly in families of Pashtun origin.

Comparative study of environmental pollutants bisphenol A and bisphenol S on sexual differentiation of anteroventral periventricular nucleus and spermatogenesis.

BACKGROUND: Bisphenol A is well known endocrine-disrupting chemical while Bisphenol S was considered a safe alternative. The present study aims to examine the comparative effects of xenobiotic bisphenol-A (BPA) and its substitute bisphenol-S (BPS) on spermatogenesis and development of sexually dimorphic nucleus population of dopaminergic neurons in the anteroventral periventricular nucleus (AVPV) of the hypothalamus in male pups. METHODS: Sprague Dawley rat's pups were administered subcutaneously at the neonatal stage from postnatal day PND1 to PND 27. Thirty animals were divided into six experimental groups (6 animals/group). The first group served as control and was provided with normal olive oil. The four groups were treated with 2 µg/kg and 200 µg/kg of BPA and BPS, respectively. The sixth group was given with 50 µg/kg of estradiol dissolved in olive oil as a standard to find the development of dopaminergic tyrosine hydroxylase neurons in AVPV regions. Histological analysis for testicular tissues and immunohistochemistry for brain tissues was performed. RESULTS: The results revealed adverse histopathological changes in testis after administration of different doses of BPA and BPS. These degenerative changes were marked by highly significant (p < 0.001) decrease in tubular and luminal diameters of seminiferous tubule and epithelial height among bisphenols treated groups as compared to control. Furthermore, significantly increased (p < 0.001) TH-ir cell bodies in the AVPV region of the brain with 200 µg/kg dose of BPA and BPS was evident. CONCLUSION: It is concluded that exposure of BPA and BPS during a critical developmental period can structural impairments in testes and affects sexual differentiation of a dimorphic dopaminergic population of AVPV region of hypothalamus in the male brain.

A novel mutation in the HPGD gene causing primary hypertrophic osteoarthropathy with digital clubbing in a Pakistani family.

Primary hypertrophic osteoarthropathy (PHO) is a congenital multisystemic entity characterized by three major clinical symptoms: pachydermia, periostosis, and digital clubbing. Recently it has been reported that pathogenic mutations in two genes are known to be associated with PHO: HPGD and SLCO2A1. In the present study, a five-generation consanguineous Pakistani family harboring primary hypertrophic osteoarthropathy in autosomal-recessive pattern was ascertained. Whole genome single nucleotide polymorphisms (SNPs) genotyping and sequence analysis revealed a novel homozygous missense mutation (c.577T˃C) of the human HPGD gene in all affected members of the family. The study presented here demonstrate the first case of primary hypertrophic osteoarthropathy reported in Pashtun population.

CTX-M-15 and OXA-10 beta lactamases in multi drug resistant Pseudomonas aeruginosa: First report from Pakistan.

BACKGROUND: Pseudomonas aeruginosa is an emerging threat to public health worldwide due to their rapid development of drug resistance including beta-lactamases. The present study was designed to investigate the incidence of ß-lactamases and genotypic pattern of CTX and OXA in the clinical isolate of multidrug resistant P. aeruginosa. METHODS: In this study a total of 102 MDR P. aeruginosa isolates obtained from Lady Reading Hospital, Peshawar, Pakistan were subjected to extended spectrum beta lactamase (ESBL), metallo beta lactamase (MBL) and plasmid mediated ß-lactamase (AmpC) detection using phenotypic and molecular methods. Furthermore, sequencing of CTX and OXA gene was performed. RESULTS: Out of 102 MDR P. aeruginosa isolates, 71 (69.6%) were beta lactamase producers. The incidence of ESBL, MBL and AmpC in clinical isolates of P. aeruginosa was found to be 23.94%, 40.84% and 35.21% respectively. Co-production of ESBL and AmpC were also observed in some isolates. There were 14 (19.71%) CTX-M-15 harboring isolates which were ESBL (64.28%), MBL (21.42%) and AmpC (14.28%) producer. Co-production of ESBL/MBL (14.28%), ESBL/AmpC (14.28%) and MBL/AmpC (14.28%) were also observed in the CTX M-15 harboring isolates while 12.28% isolates were not ESBL, MBL or AmpC producer. OXA-10 was detected in 8 (11.26%) isolates which were ESBL (12.5%), MBL (37.5%) and AmpC (12.5%) producer. OXA 10 isolates also exhibit co-production of ESBL/AmpC (12.5%) and MBL/AmpC (12.5%). All CTX-M-15 carried the class A ß-lactamase conserved domain while OXA-10 harbored conserved domain of class D ß-lactamase. CONCLUSION: The current study for the first time reported and characterized the CTX-M-15 and OXA-10 among MDR P. aeruginosa isolates from Pakistan. Further efforts are needed to understand the molecular mechanism of drug resistance with CTX and OXA harboring P. aeruginosa isolates.

Expanding the clinical and genetic spectrum of G6PD deficiency: The occurrence of BCGitis and novel missense mutation.

Glucose-6-phosphate dehydrogenase (G6PD) is a key enzyme in the pentose phosphate pathway that ensures sufficient production of coenzyme nicotinamide adenine dinucleotide phosphate (NADPH) by catalyzing the reduction of NADP+ to NADPH. Noteworthy, the latter mediates the production of reactive oxygen species (ROS) by phagocytic cells such as neutrophils and monocytes. Therefore, patients with severe forms of G6PD deficiency may present impaired NADPH oxidase activity and become susceptible to recurrent infections. This fact, highlights the importance to characterize the immunopathologic mechanisms underlying the susceptibility to infections in patients with G6PD deficiency. Here we report the first two cases of G6PD deficiency with Bacille Calmette-Guérin (BCG) adverse effect, besides jaundice, hemolytic anemia and recurrent infections caused by Staphylococcus aureus. The qualitative G6PD screening was performed and followed by oxidative burst analysis using flow cytometry. Genetic and in silico analyses were carried out by Sanger sequencing and mutation pathogenicity predicted using bioinformatics tools, respectively. Activated neutrophils and monocytes from patients displayed impaired oxidative burst. The genetic analysis revealed the novel missense mutation c.1157T>A/p.L386Q in G6PD. In addition, in silico analysis indicated that this mutation is pathogenic, thereby hampering the oxidative burst of neutrophils and monocytes from patients. Our data expand the clinical and genetic spectrum of G6PD deficiency, and suggest that impaired oxidative burst in this severe primary immune deficiency is an underlying immunopathologic mechanism that predisposes to mycobacterial infections.

Multi drug resistant Pseudomonas aeruginosa: Pathogen burden and associated antibiogram in a tertiary care hospital of Pakistan.

Pseudomonas aeruginosa is an important pathogen of both community and hospital acquired infections, and a major threat to public health for continuous emergence of multi-drug resistance. Current prevalence and pattern of multidrug resistance in the clinical isolates of P. aeruginosa is reported here. Samples were collected from September 2013 to January 2014 tertiary care hospital, Peshawar. Samples were subjected to phenotypic and molecular based detection of P. aeruginosa and were further processed for multidrug resistance pattern. Out of 3700 samples, 102 were identified as MDR P. aeruginosa. Prevalence of MDR isolates were found in pus (34.3%), wounds (28.4%), urine (19.6%), blood (14.7%) and sputum (2.9%) respectively. Isolates were more resistant to Sulphamethoxazole/Trimethoprim (98.04%), Amoxycillin/Clavulanic acid, Doxycycline and Chloramphenicol (95.1%) each, while least resistant to Imipenem (43.1%), Cefoperazone/Sulbactam (50.98%) and Amikacin (53.9%). Extensive MDR pattern was observed in P. aeruginosa was found as (n = 17, 16.6%) isolates were resistant to all four classes of antibiotics. Increased burden of MDR P. aeruginosa was documented in the study. Moreover, some isolates were even resistant to four classes of antibiotics. Findings of the study will be helpful to devise an appropriate antibiotic treatment strategy against MDR P. aeruginosa to cope the chances of evolving resistant pathogens.

Phenotypic manifestation of congenital transverse amputation of autopod in Pakistani subjects.

Terminal transverse deficiency of forearm is a very rare limb malformation. Most of the cases have traumatic etiology and congenital presentation is less common. A series of six individuals with transverse deficiency through the hands is presented in this communication. The cases were congenital, morphologically similar and showed loss of four fingers, most often postaxial. The affected arm was reduced in size compared to the contralateral limb and there was distortion of palmer creases. All cases were sporadic and non-syndromic in nature. The characteristics of these cases were concordant with the symbrachydactyly type III or monodactylous type, when classified according to the scheme proposed by Blauth and Gekeler (1973). The malformation resulted in permanent quality-of-life impairment in these subjects and warrant prosthetic management. Detailed physical and phenotypic features of the patients have been presented.

Chlorophyll derivatives can be an efficient weapon in the fight against dengue.

Dengue, a mosquito-borne viral infection, is one of the major public health concerns in the tropical and subtropical regions of the world. Approximately, 2.5 billion people across the world are at risk from dengue and 50 to 100 million new infections of dengue occur annually. There is yet no vaccine or medicine available against dengue, and treatment remains only supportive. Targeting its vector by a combination of biological and chemical approaches and management of breeding sites are currently the only existing approaches to control or eliminate dengue. Chlorophyll derivatives like chlorophyllin and pheophorbide have been reported as effective natural photosensitizers against larvae of several insects including flies. Chlorophyll derivatives were also reported effective against larval stages of freshwater snails as well as against certain parasites of fish. This article briefly discusses the possible application of chlorophyll derivatives in controlling dengue vectors and hence the disease itself. Chlorophyll derivatives can prove to be a good contributor in an integrated approach against dengue.

Augmented human thermal discomfort in urban centers of the Arabian Peninsula.

Anthropogenic climate change has amplified human thermal discomfort in urban environments. Despite the considerable risks posed to public health, there is a lack of comprehensive research, evaluating the spatiotemporal changes in human thermal discomfort and its characteristics in hot-hyper arid regions, such as the Arabian Peninsula (AP). The current study analyzes spatiotemporal changes in human thermal discomfort categories and their characteristics in AP, using the newly developed high-resolution gridded ERA5-HEAT (Human thErmAl comforT) dataset for the period 1979-2022. In addition, the study assesses the interplay between the Universal Thermal Climate Index (UTCI) and El Niño-Southern Oscillation (ENSO) indices for the study period. The results reveal a significant increase in human thermal discomfort and its characteristics, with higher spatial variability in the AP region. The major urban centers in the southwestern, central, and southeastern parts of AP have experienced significant increases in human thermal discomfort (0.4-0.8 °C), with higher frequency and intensity of thermal stress during the study period. The temporal distribution demonstrates a linear increase in UTCI indices and their frequencies and intensities, particularly from 1998 onward, signifying a transition towards a hotter climate characterized by frequent, intense, and prolonged heat stress conditions. Moreover, the UTCI and ENSO indices exhibit a dipole pattern of correlation with a positive (negative) pattern in the southwestern (eastern parts) of AP. The study's findings suggest that policymakers and urban planners need to prioritize public health and well-being in AP's urban areas, especially for vulnerable groups, by implementing climate change adaptation and mitigation strategies, and carefully designing future cities to mitigate the effects of heat stress.

Uncertainty reduction for precipitation prediction in North America.

Large differences in projected future annual precipitation increases in North America exists across 27 CMIP6 models under four emission scenarios. These differences partly arise from weak representations of land-atmosphere interactions. Here we demonstrate an emergent constraint relationship between annual growth rates of future precipitation and growth rates of historical temperature. The original CMIP6 projections show 0.49% (SSP126), 0.98% (SSP245), 1.45% (SSP370) and 1.92% (SSP585) increases in precipitation per decade. Combining observed warming trends, the constrained results show that the best estimates of future precipitation increases are more likely to reach 0.40-0.48%, 0.83-0.93%, 1.29-1.45% and 1.70-1.87% respectively, implying an overestimated future precipitation increases across North America. The constrained results also are narrow the corresponding uncertainties (standard deviations) by 13.8-31.1%. The overestimated precipitation growth rates also reveal an overvalued annual growth rates in temperature (6.0-13.2% or 0.12-0.37°C) and in total evaporation (4.8-14.5%) by the original models' predictions. These findings highlight the important role of temperature for accurate climate predictions, which is important as temperature from current climate models' simulations often still have systematic errors.

Analysis of molecular subtypes and antibiotic resistance in Treponema pallidum isolates from blood donors in Khyber Pakhtunkhwa, Pakistan.

Syphilis, caused by Treponema pallidum, is resurging globally. Molecular typing allows for the investigation of its epidemiology. In Pakistan and other nations, T. pallidum subsp. pallidum has developed widespread macrolide resistance in the past decade. A study at the Peshawar Regional Blood Centre from June 2020-June 2021 analyzed serum samples from 32,812 blood donors in Khyber Pakhtunkhwa, Pakistan, to assess circulating T. pallidum strains and antibiotic resistance. Blood samples were initially screened for T. pallidum antibodies using a chemiluminescent microparticle immunoassay (CMIA). CMIA-reactive samples underwent polymerase chain reaction (PCR) targeted the polA, tpp47, bmp, and tp0319 genes. PCR-positive samples were further analyzed for molecular subtyping using a CDC-developed procedure and tp0548 gene examination. All PCR-positive samples were analyzed for the presence of point mutations A2058G and A2059G in 23S rRNA, as well as the G1058C mutation in 16S rRNA. These mutations are known to impart antimicrobial resistance to macrolides and doxycycline, respectively. Out of 32,812 serum samples, 272 (0.83%) were CMIA-reactive, with 46 being PCR-positive. Nine T. pallidum subtypes were identified, predominantly 14d/f. The A2058G mutation in 23S rRNA was found in 78% of cases, while G1058C in 16S rRNA and A2059G in 23S rRNA were absent. The research found donor blood useful for assessing T. pallidum molecular subtypes and antibiotic resistance, especially when chancres are not present. The prevalent subtype was 14d/f (51.85%), and the high macrolide resistance of 36 (78%) indicates caution in using macrolides for syphilis treatment in Khyber Pakhtunkhwa, Pakistan.

Heavy metal impacts on antioxidants in cow blood from wastewater-irrigated areas.

The aim of the present research was to investigate the presence of heavy metals such as lead (Pb), copper (Cu), chromium (Cr), and cadmium (Cd) in blood samples from cows raised with irrigated wastewater, as well as in the wastewater itself, in the North-western region of Pakistan. A total of 60 blood samples were collected from five different locations in Kohat, namely Tappi Road (TR), Pindi Road (PR), Gul Malik Road (GMR), Markaz Road (MR), and a control group. The samples of both i.e. cow blood and wastewater were analyzed for the concentrations of heavy metals. The highest concentration of Cd was detected in the MR site with a mean value of 0.03 mg/L, and the highest concentration of Cu (0.04 mg/L) was recorded in the TR site, while the lowest level was found in the control group with a mean of 0.002 mg/L in blood samples. The highest Cr and Pb concentrations were found at the PR site, with mean values of 0.03 and 0.07 mg/L, respectively, whereas the control group had the lowest concentrations, with mean values of 0.002 and 0.01 mg/L. Similarly, heavy metal concentrations were analyzed in wastewater used for irrigation in the study area. Results indicated elevated concentrations of Cu and Cr in wastewater, although they remained below the World Health Organization (WHO) recommended values except for Cr (0.13 mg/L) in the GMR site, which exceeded permissible limits. Cd and Pb concentrations in wastewater were relatively low, but Cd concentration surpassed WHO limits, particularly with a mean concentration of 0.08 mg/L in the TR site. Comparison between heavy metal concentrations in blood and wastewater revealed higher values of Cd and Pb in blood samples than in wastewater, while Cu and Cr concentrations were higher in water compared to blood. Additionally, elevated levels of Super Oxide Dismutase (SOD), antioxidant enzyme Catalase (CAT), and oxidative stress marker malondialdehyde (MDA) were detected in blood samples. Cluster and principal component analyses were employed to assess heavy metal toxicity among the groups, indicating potential long-term adverse health effects on animals, transfer to humans, and toxicity in living organisms.

The detrimental effects of heavy metals on tributaries exert pressure on water quality, Crossocheilus aplocheilus, and the well-being of human health.

The escalating presence of heavy metals (HMs) in the Panjkora River water and their impact on fish pose a significant challenge to both the ecological community and human health. Consequently, a study was conducted with the primary aim of elucidating their influence on human health-related issues. To address this, the concentrations of heavy metals, including arsenic (As), cadmium (Cd), iron (Fe), manganese (Mn), lead (Pb), and zinc (Zn), in both water and the fish species Crossocheilus diplocheilus were investigated across various locations within the study area. The quantification of HMs concentration was carried out utilizing an atomic absorption spectrophotometer. The highest concentration in water was found as 0.060 mg/L for Pb and lowest for Fe, whereas the highest concentration in fish was 2.028 mg/kg for Pb and lowest for As. Human health risk associated with fish eating was evaluated by using health risk indices (HRI) for non-carcinogenic health risks and targeted cancer risk (TR) for carcinogenic health risks. The values of the health risk index (HRI) were found greater than 1 except Fe (0.0792), Zn (0.782), and Mn (0.541). The highest mean HRI > 1 was recorded for As (62.99), Cd (26.85), and Pb (10.56). This implies that fish consumption from river Panjkora is not safe up to some extent. Similarly, the TR value for As, Cd, and Pb was found 2.8 [Formula: see text], 1.6 [Formula: see text], 2.8 ×[Formula: see text] which showed cancer risk. There is a detected risk to human health associated with the consumption of fish from the Panjkora River. The government must implement adaptive measures to address this significant issue of water pollution in the study area. Additionally, there is a need for further extensive and prolonged research studies in this context.

Ecotoxicity evaluation of a liquid detergent using the automatic biotest ECOTOX.

Synthetic detergents are common pollutants reaching aquatic environments in different ways after usage at homes, institutions and industries. In this study a liquid detergent, used for dish washing, was evaluated for its toxicity during long- and short-term tests using the automatic biotest ECOTOX. Different parameters of Euglena gracilis like motility, swimming velocity, gravitactic orientation, cell compactness and cell growth were used as end points. In short-term experiments, the maximum adverse effects on motility, velocity, cell shape and gravitaxis were observed after 1 h of exposure. With further increase in exposure time to the detergent a slight recovery of these parameters was observed. In long-term experiments, the detergent caused severe disturbances to E. gracilis. Motility, cell growth and cell compactness (shape) with EC50 values of 0.064, 0.18 and 2.05 %, respectively, were found as the most sensitive parameters to detergent stress. There was a slight positive effect on gravitactic orientation at the lowest two concentrations; at higher concentrations of the detergent cells orientation was highly impaired giving EC50 values of 1.75 and 2.52 % for upward swimming and r-value, respectively.

Proteomic map of the differentially expressed proteins in the skin of Ctenopharyngodon idella against Aeromonas hydrophila infection.

The skin mucus of fish is an important part of the innate immune system, which is poorly understood at the proteomic level. The study established a complete map of the proteins in the skin mucus of Ctenopharangdon idella (C. idella) and discussed the Differentially Expressed Proteins (DEPs) after Aeromonas hydrophila (A. hydrophila) infection. Using Label Free Liquid Chromatography-Mass Spectrometry (LC-MS/MS) analysis, a total of 126 proteins were identified as differentially expressed, 89 proteins of which were upregulated, and 37 proteins were downregulated. Functional annotations of DEPs showed that the upregulated proteins in the skin mucus of the treated group were mostly associated with complement system and cytoskeleton proteins, whereas downregulated proteins were associated with metabolism. The key upregulated immune proteins were transferrin variant C, lysozyme g, annexin A11, 26S proteasome non-ATPase regulatory subunit 8, hypothetical protein ROHU_000884, 60S ribosomal L7a, calpain-2 catalytic subunit-like protein, calpain-9-like protein, complement component C9, complement C3, cathepsin S, cathepsin Z, 14 kDa apolipo, heat shock protein and intelectin, whereas, leukocyte elastase inhibitor, annexin A11, C-factor-like protein, biotinidase isoform X1 and epidermal growth factor receptor substrate 15-like were the downregulated proteins. Moreover, we for the first-time report proteins such as coactosin, lamin-B2 and kelch 12, which were never reported in fish. Our study directly pointing out the possible immunological biomarkers in the skin mucus of C. idella after A. hydrophila treatment. Each of the protein we report in this study could be used as base to establish their mechanism of action during bacterial infection that may contribute to the strategies against bacterial prevention and control in fishes.