Knowledge, Attitude, and Perception of Cancer Patients towards COVID-19 in Pakistan: A Cross-Sectional Study.

Background: Cancer patients, being immunocompromised, are at higher risk of coronavirus disease (COVID-19). The current study determines cancer patients' knowledge, attitude, perception, and impact of the COVID-19 pandemic. Method: A cross-sectional online survey was conducted in Pakistan from 1 April 2020 to 1 May 2020. The study respondents were cancer patients with ages equal to or greater than 18 years. Following a request for participation, the URL for the survey was distributed on numerous channels. Other social media platforms, including WeChat, WhatsApp, Facebook, Twitter, Instagram, Messenger, and LinkedIn, were used to increase cancer patient interaction. The questionnaire comprised five different sections such as: (1) sociodemographic information, (2) knowledge, (3) attitude, (4) perception, and (5) impact of COVID-19 on cancer patients. Descriptive medical statistics such as frequency, percentage, mean, and standard deviation were used to illustrate the demographic characteristics of the study participants. To compare mean knowledge scores with selected demographic variables, independent sample t-tests and one-way analysis of variance (ANOVA) were used, which are also practical methods in epidemiological, public health and medical research. The cut-off point for statistical significance was set at a p-value of 0.05. Results: More than 300 cancer patients were invited, of which 208 agreed to take part. The response rate was 69.33% (208/300). Gender, marital status, and employment status had a significant association with knowledge scores. Of the total recruited participants, 96% (n = 200) (p < 0.01) knew about COVID-19, and 90% were aware of general symptoms of COVID-19 disease, such as route of transmission and preventive measurements. In total, 94.5% (n = 197) (p < 0.01) were willing to accept isolation if they were infected with COVID-19, and 98% (n = 204) (p < 0.01) had reduced their use of public transportation. More than 90% (n = 188) (p < 0.01) of cancer patients were found to be practicing preventative measures such as using a face mask, keeping social distance, and avoiding handshaking and hugging. Around 94.4% (n = 196) (p < 0.01) of cancer patients had been impacted by, stopped or had changed cancer treatment during this pandemic, resulting in COVID-related anxiety and depression. Conclusion: The included cancer patients exhibited a good level of COVID-19 knowledge, awareness, positive attitude, and perception. Large-scale studies and efforts are needed to raise COVID-19 awareness among less educated and high-risk populations. The present survey indicates that mass-level effective health education initiatives are required for developing countries to improve and reduce the gap between KAP and COVID-19.

Quantitative Estimation of Aflatoxin Level in Poultry Feed in Selected Poultry Farms.

Statement of Novelty. Poultry feed contamination due to mycotoxins is one of the major threats to the growing poultry industry. Surveillance of different mycotoxins, including aflatoxin, is very important to control economic and health hazards associated with these toxins. Studies reporting aflatoxin levels in poultry feed are limited. Therefore, this study was conducted to examine the occurrence of total aflatoxin in poultry feed. This study is the first-ever documentation about the frequency and quantitative estimations of total aflatoxin levels in poultry feed consumed to provide solid feedback to the poultry industrialists and researchers involved in studying the mycotoxins. Objective. Contamination of poultry feed with mycotoxins such as aflatoxin is a major concern for the poultry industry that results in a significant economic loss and directly affects consumers. Monitoring the aflatoxin levels in poultry feed is crucial for controlling economic loss and decreasing the health hazards to the population. This study was conducted to examine the occurrence of total aflatoxin in poultry feed in a high consumption area. Three different poultry feeds, i.e., starter, grower, and finisher, were assessed through continuous sampling from farms. The incidence of positive samples for aflatoxin contamination was 92.5%. Grower feed had the highest frequency (100%) of aflatoxin positive samples and aflatoxin levels with a mean value of 56.34 ppb. Further, the range of moisture content was around 6.8%-10.98%. No significant correlation between humidity and aflatoxin contamination was revealed when analyzed by Pearson's correlation coefficient with r 2 of 0.05 and p value of 0.13. The results warrant the need for constant monitoring programs for the prevention of aflatoxin contamination in local poultry farms.

Preparation, characterization and stability studies of cross-linked α-amylase aggregates (CLAAs) for continuous liquefaction of starch.

In current study, α-amylase of fungal origin was immobilized using cross-linking strategy. The influence of precipitant (ammonium sulphate) and cross-linker (glutaraldehyde) concentration revealed that 60% (w/v) precipitant and 1.5% (v/v) cross-linker saturation was required to attain optimum activity. Cross-linked amylase aggregates (CLAAs) were characterized and 10-degree shift in optimum temperature (soluble enzyme: 50 °C; cross-linked: 60 °C) and 1-unit shift in pH (soluble enzyme: pH -6; cross-linked: pH -7) was observed after immobilization. The Vmax for soluble α-amylase and its cross-linked form was 1225 U ml-1 and 3629 U ml-1, respectively. The CLAAs was more thermostable than its soluble form and retained its 30% activity even after 60 min of incubation at 70 °C. Moreover, cross-linked amylase retained its activity after two months while its soluble counterpart lost its complete activity after 10 and 20 days at 30 °C and 4 °C storage, respectively. Reusability test showed that cross-linked amylase could retain 13% of its residual activity after 10 repeated cycles. Therefore, 10 times more glucose was produced after cross-linking than soluble amylase when it was utilized multiple times. This study indicates that amylase aggregates are highly effective for continuous liquefaction of starch, hence have strong potential to be used for different industrial processes.

Mycoremediation: a treatment for heavy metal-polluted soil using indigenous metallotolerant fungi.

Bioleaching of heavy metals from industrial contaminated soil using metallotolerant fungi is the most efficient, cost-effective, and eco-friendly technique. In the current study, the contaminated soil samples from Hattar Industrial Estate revealed a total lead (Pb) and mercury (Hg) concentration of 170.90 mg L-1 and 26.66 mg L-1, respectively. Indigenous metallotolerant fungal strains including Aspergillus niger M1, Aspergillus fumigatus M3, Aspergillus terreus M6, and Aspergillus flavus M7 were isolated and identified by pheno- and genotyping. A. fumigatus and A. flavus of soil sample S1 showed higher efficiency for Pb removal (99.20% and 99.30%, respectively), in SDB medium. Likewise, A. niger and A. terreus of soil sample S2 showed higher efficiency for Hg removal (96% and 95.50%, respectively), in YPG medium. Furthermore, the maximum uptake efficiency for Pb removal (8.52 mg g-1) from soil sample S1 was noticed for A. fumigatus in YPG medium, while the highest uptake efficiency (4.23 mg g-1) of A. flavus M2 strain was observed with CYE medium. Similarly, the maximum uptake efficiency of 0.41 mg g-1 and 0.44 mg g-1 for Hg removal from soil sample S2 was found for A. niger and A. terreus strains, respectively, in CYE medium. Thus, in order to address the major issue of industrial waste pollution, indigenous fungal strains A. fumigatus (M1) and A. terreus (M7), isolated in this study, could be used (ex situ or in situ) to remediate soils contaminated with Pb and Hg.

Mycoremediation of heavy metal (Cd and Cr)-polluted soil through indigenous metallotolerant fungal isolates.

Remediation of heavy metals, other than microbial bioleaching method, is expensive and unsuitable for large contaminated areas. The current study was aimed to isolate, identify, and test the potential of indigenous fungal strains for heavy metal removal from contaminated soil. A total of three metallotolerant fungal strains, i.e., Aspergillus niger (M1DGR), Aspergillus fumigatus (M3Ai), and Penicillium rubens (M2Aii), were isolated and identified by phenotyping and genotyping from heavy metal-contaminated soil of  Hattar Industrial Estate, Pakistan. A. niger was found to be the most successful strain for the removal of heavy metals from the contaminated soil with maximum bioaccumulation efficiency of 98% (Cd) and 43% (Cr). In contrast, A. fumigatus showed comparatively low but still considerable bioleaching potential, i.e., 79% and 69% for Cd and Cr removal, respectively. Maximum metal uptake efficiency, i.e., 0.580 mg g-1 and 0.152 mg g-1 by A. niger strain was noticed for Cd and Cr with Czapek yeast extract (CYE) and Sabouraud dextrose broth (SDB) media, respectively. A. fumigatus (M3Ai) exhibited the maximum bioleaching capacity (0.40 mg g-1) for Cr with CYE medium. The results reveal that A. niger M1DGR and A. fumigatus M3Ai could be used to develop new strategies to remediate soil contaminated with heavy metals (Cd and Cr) through either in situ or ex situ mycoremediation.

Isolation, identification, and pathological effects of beach sand bacterial extract on human skin keratinocytes in vitro.

BACKGROUND: Beaches are recreational spots for people. However, beach sand contains harmful microbes that affect human health, and there are no established methods for either sampling and identifying beach-borne pathogens or managing the quality of beach sand. METHOD: This study was conducted with the aim of improving human safety at beaches and augmenting the quality of the beach experience. Beach sand was used as a resource to isolate bacteria due to its distinctive features and the biodiversity of the beach sand biota. A selected bacterial isolate termed FSRS was identified as Pseudomonas stutzeri using 16S rRNA sequencing and phylogenetic analysis, and the sequence was deposited in the NCBI GenBank database under the accession number MF599548. The isolated P. stutzeri bacterium was cultured in Luria-Bertani growth medium, and a crude extract was prepared using ethyl acetate to examine the potential pathogenic effect of P. stutzeri on human skin. A human skin keratinocyte cell line (HaCaT) was used to assess cell adhesion, cell viability, and cell proliferation using a morphological analysis and a WST-1 assay. RESULT: The crude P. stutzeri extract inhibited cell adhesion and decreased cell viability in HaCaT cells. We concluded that the crude extract of P. stutzeri FSRS had a strong pathological effect on human skin cells. DISCUSSION: Beach visitors frequently get skin infections, but the exact cause of the infections is yet to be determined. The beach sand bacterium P. stutzeri may, therefore, be responsible for some of the dermatological problems experienced by people visiting the beach.

Cloning and characterization of pyruvate carboxylase gene responsible for calcium malate overproduction in Penicillium viticola 152 and its expression analysis.

In this study, a pyruvate carboxylase gene (PYC) from a marine fungus Penicillium viticola 152 isolated from marine algae was cloned and characterized by using Genome Walking method. An open reading frame (ORF) of The PYC gene (accession number: KM593097) had 3582bp encoding 1193 amino acid protein (isoelectric point: 5.01) with a calculated molecular weight of 131.2757kDa. A putative promoter (intronless) of the gene was located at -666bp and contained a TATA box, several CAAT boxes, the 5'-SYGGRG-3' and a 5'-HGATAR-3' sequences. A consensus polyadenylation site (AATAAA) was also observed at +10bp downstream of the ORF. The protein deduced from the PYC gene had no signal peptide, was a homotetramer (4), and had the four functional domains. Furthermore, PYC protein also had three potential N-linked glycosylation sites, among them, -N-S-T-I- at 36 amino acid, -N-G-T-V- at 237 amino acid, and -N-G-S-S- at 517 amino acid were the most possible N-glycosylation sites. After expression of the PYC gene of P. viticola 152 in medium supplemented with CSL and biotin, it was found that the specific pyruvate carboxylase activity in MA production medium supplemented with CSL was much higher (0.5U/mg) than in MA medium supplemented with biotin (0.3U/mg), suggesting that optimal concentration of CSL is required for increased expression of the PYC gene, which is responsible for high level production of malic acid in P. viticola 152 strain.