Assessment of natural radioactivity levels in rice samples and their implications for radiological protection.

This study investigates the levels of natural and artificial radioactivity in rice samples collected from various local markets in Islamabad, Pakistan. The 226Ra, 232Th, and 40K activity concentrations were measured through gamma-ray spectrometry with a NaI(Tl) detector. The average activity concentrations were 1.67 ± 1.19 Bq kg-1 for 226Ra, 3.31 ± 1.83 Bq kg-1 for 232Th, and 88.51 ± 11.65 Bq kg-1 for 40K. Calculated radium equivalent (Raeq) values ranged from 7.35 to 18.08 Bq kg-1, with a mean value of 11.11 Bq kg-1, all below the permissible maximum of 370 Bq kg-1. The absorbed dose rates ranged from 6.85 to 16.39 nGy h-1, with an average of 10.64 nGy h-1, falling below the acceptable limit of 51 nGy h-1. The outdoor and indoor radiation hazard indices (Hex and Hin) had mean values of 0.03, both below the threshold value of one. The external and internal hazard indices (Iγ and Iα) were both 0.088, also below the critical value of one. The excess lifetime cancer risk (ELCR) ranged from 0.28 to 0.11, with a mean value of 0.18, which is less than the critical value of one. Overall, the radioactivity levels in the analyzed rice samples are within the acceptable limits set by the International Commission on Radiological Protection and are below global averages. These results offer important insights into the radiological safety of rice consumption in the study area.

Seed priming with Fe3O4-SiO2 nanocomposites simultaneously mitigate Cd and Cr stress in spinach (Spinacia oleracea L.): A way forward for sustainable environmental management.

Seed priming with a composite of iron oxide (Fe3O4) and silicon dioxide (SiO2) nanoparticles (NPs) is an innovative technique to mitigate cadmium (Cd) and chromium (Cr) uptake in plants from rooting media. The current study explored the impact of seed priming with varying levels of Fe3O4 NPs, SiO2 NPs, and Fe3O4-SiO2 nanocomposites on Cd and Cr absorption and phytotoxicity, metal-induced oxidative stress mitigation, growth and biomass yield of spinach (Spinacia oleracea L.). The results showed that seed priming with the optimum level of 100 mg L-1 of Fe3O4-SiO2 nanocomposites significantly (p ≤ 0.05) increased root dry weight (144 %), shoot dry weight (243 %) and leaf area (34.4 %) compared to the control, primarily by safeguarding plant's photosynthetic machinery, oxidative stress and phytotoxicity of metals. Plants treated with this highest level of Fe3O4-SiO2 nanocomposites exhibited a substantial increase in photosynthetic and gas exchange indices of spinach plants and enhanced activities of superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX) antioxidant enzymes by 45 %, 48 %, and 60 %, respectively. Correspondingly, the relative gene expression levels of SOD, CAT, and APX also rose by 109 %, 181 %, and 137 %, respectively, compared to non-primed plants. This nanocomposite application also boosted the levels of phenolics (28 %), ascorbic acid (68 %), total sugars (129 %), flavonoids (39 %), and anthocyanin (29 %) in spinach leaves, while significantly reducing Cd (34.7 %, 53.4 %) and Cr (20.2 %, 28.8 %) contents in plant roots and shoots, respectively. These findings suggest that seed priming with Fe3O4-SiO2 nanocomposites effectively mitigated the toxic effects of Cd and Cr, enhancing the growth and biomass yield of spinach in Cd and Cr co-contaminated environments, offering a promising sustainable approach for producing metal-free crops.

Microwave-sonication synergistic extraction of dairy waste proteins: A review of green approach for dairy waste proteins valorization.

Ultrasonic and microwave extraction process has great prospects to convert food and agricultural waste from food industries to value-added goods. Also, this review extensively elaborates the utilization of ultrasonication and microwave extraction (US-MW) process for valorization of dairy waste extracted proteins into novel foods. Both of these extraction and processing techniques are considered as green technologies when compared with the other conventional or chemical extraction and processing techniques. Further, this review also explains the impact of US-MW alone and in combination on the dairy waste proteins extraction, nutritional and techno-functional attributes of these dairy-waste proteins. The review also highlights the economic and cost-effective benefits of US-MW processes for extracting the proteins from dairy waste, indicating their feasibility and sustainability. The review also elucidated the synergistic utilization of US-MW extraction as a viable processing technique in extraction or production of bioactive compounds like dairy proteins. In conclusion, this review elucidates the US-MW, both individually and in synergy as a viable source of dairy waste proteins extraction and their application in functional foods. Moreover, in accordance to the latest developments and future prospects at pilot and commercial level to assess the practicability of synergistic use of US-MW extraction in bioenergy production from food wastes other than dairy waste for extraction and production of biodiesel, hydrogen, green methane, and ethanol.

Revolutionizing the food industry: The transformative power of artificial intelligence-a review.

Artificial Intelligence (AI) is revolutionizing the food industry by optimizing processes, improving food quality and safety, and fostering innovation. This review examines AI's applications in food science, including supply chain management, production, sensory science, and personalized nutrition. It discusses techniques like knowledge-based expert systems, fuzzy logic, artificial neural networks, and machine learning, highlighting their roles in predictive maintenance, quality control, product development, and waste management. The integration of AI with sophisticated sensors enhances real-time monitoring and decision-making in food safety and packaging. However, challenges such as ethical concerns, data security, transparency, and high costs persist. AI is poised to advance sustainability by optimizing resource use, enhance food security through predictive analytics of crop yields, and drive innovation in personalized nutrition and supply chain automation, ensuring tailored products and efficient delivery. This paper underscores AI's transformative potential in the food industry while addressing the obstacles to its widespread adoption.

Ultrasound-assisted extraction of bioactive compounds from Moringa oleifera leaves for beef patties preservation: Antioxidant and inhibitory activities, half-life, and sensory attributes.

This study aims to examine employing ultrasound-assisted extraction of bioactive components from Moringa oleifera leaves and apply them in beef patties preservation, as well as antioxidant and inhibitory activities and sensory qualities. The study included studying the chemical content and minerals of the M. oleifera leaves, preparation of aqueous and alcoholic extracts using an ultrasound device, then exploring the extraction yield. The results proved that the extraction yield by ultrasound using ethanol at 80% was the highest, reaching 19.22%. The total phenols in the ultrasonic extract with moringa leaves aqueous extract (AMEUS) amounted to 120,755 mg/mL. Since the AMEUS exhibited the highest value of 68.308 mg/mL calic acid - eight phenolic compounds discovered by HPLC - the total content of flavonoids was also calculated. The inhibitory and antioxidant effects of moringa leaf extracts are well documented. We monitored the changes in chemical indicators, such as the value of peroxide and thiobarbituric acid, as well as the percentage of free fatty acids and physical characteristics, such as water-carrying capacity, pH, and pigments, for storage periods 0, 4, 8, and 12 days after adding AMEUS to beef patties at a concentration of 0.5%. The patties were kept under refrigeration at 4 ± 1°C during this time. The values of peroxide number, thiobarbituric acid, free fatty acid, and metmyoglobin pigment were decreased in the beef patties treated with the AMEUS. However, they increased continuously during the cryopreservation period, and there was a significant increase in water-holding capacity (WHC) when the beef patties were treated with AMEUS. The results also showed that adding AMEUS to beef patties improved their qualitative characteristics.

Modulation of Cd carriers by innovative nanocomposite (Ca+Mg) and Cd-resistance microbes (Bacillus pumilus): a mechanistic approach to enhance growth and yield of rice (Oryza sativa L.).

Cadmium (Cd) is a well-known pollutant in agricultural soil, affecting human health through the food chain. To combat this issue, Ca + Mg (25 mg L-1) nanocomposite and Bacillus pumilus, either alone or combined, were applied to rice plants under Cd (5 mg kg-1, 10 mg kg-1) contamination. In our study, growth and yield traits demonstrated the beneficial influence of Ca + Mg and B. pumilus application in improving rice defense mechanism by reducing Cd stress. Combined Ca + Mg and B. pumilus application increased SPAD (15), total chlorophyll (18), chlorophyll a (11), chlorophyll b (22), and carotenoids (21%) with Cd (10 mg kg-1), compared to the application alone. Combined Ca + Mg and B. pumilus application significantly regulated MDA (15), H2O2 (13), EL (10), and O2 •- (24%) in shoots under Cd (10 mg kg-1), compared to the application alone. Cd (10 mg kg-1) increased the POD (22), SOD (21), APX (12), and CAT (13%) in shoots with combined Ca + Mg and B. pumilus application, compared to the application alone. Combined Ca + Mg and B. pumilus application significantly reduced Cd accumulation in roots (22), shoots (13), and grains (20%) under Cd (10 mg kg-1), compared to the application alone. Consequently, the combined application of Ca + Mg and B. pumilus is a sustainable solution to enhance crop production under Cd stress.

Disposition and Follow-up for Low-Risk Febrile Infants: A Secondary Analysis of a Multicenter Study.

OBJECTIVE: American Academy of Pediatrics guidelines recommend that febrile infants at low risk for invasive bacterial infection be discharged from the emergency department (ED) if primary care provider (PCP) follow-up occurs within 24 hours. We aimed to (1) assess the association between having electronic health record (EHR) documentation of a PCP and ED disposition and (2) describe documentation of potential barriers to discharge and plans for post-discharge follow-up in low-risk febrile infants. METHODS: We conducted a secondary analysis of a multicenter, cross-sectional study of low-risk febrile infants. Descriptive statistics characterized ED disposition on the basis of the day of the visit, EHR documentation of PCP, scheduled or recommended PCP follow-up, and barriers to discharge. RESULTS: Most infants (3565/4042, 90.5%) had EHR documentation of a PCP. Compared with discharged infants, a similar proportion of hospitalized infants had EHR documentation of PCP (90.3% vs 91.2%, P = .47). Few infants (1.5%) had barriers to discharge documented. Of the 3360 infants (83.1%) discharged from the ED, 1544 (46.0%) had documentation of scheduled or recommended 24-hour PCP follow-up. Discharged infants with weekday visits were more likely than those with weekend visits to have documentation of scheduled or recommended 24-hour follow-up (50.0% vs 35.5%, P < .001). CONCLUSIONS: Most infants had a documented PCP, yet fewer than half had documentation of a scheduled or recommended 24-hour follow-up. A dedicated focus on determining post-ED care plans that are safe and patient-centered may improve the quality of care for this population.

Preovulatory follicular dynamics and ovulatory events following the use of GnRH 84 h after medroxyprogesterone acetate sponge removal in postpartum buffaloes.

Herein, we evaluated the effects of Gonadotropin hormone-releasing hormone (GnRH) administration 84 h after medroxyprogesterone acetate (MAP) sponge removal on follicular growth, ovulation timing, and pregnancy per artificial insemination (AI) in cosynchronized postpartum Nili Ravi buffaloes. In this study, 58 Nili Ravi postpartum buffaloes (DIM = 103 ± 1.64) were randomly divided into two treatment groups (n = 29/treatment): GnRH-TAI-84 and TAI-84. All buffaloes were administered a MAP sponge for seven days. Upon MAP sponge removal, all the subjects received prostaglandin F2α (PGF2α), and Timed AI (TAI) was performed 84 hours after sponge removal. In the GnRH-TAI-84 group, the buffaloes received GnRH alongside insemination, whereas in the TAI-84 group, the buffaloes were inseminated without GnRH administration. Follicle diameter and blood estradiol levels were measured every 6 h from 72-108 h after MAP sponge removal. The animals were checked for pregnancy using ultrasonography 40 days after AI. Animals subjected to the GnRH-TAI-84 protocol had a higher follicular growth rate and preovulatory follicle size than those in the TAI-84 group. The follicular diameter was also larger in animals that received GnRH-TAI-84 than in those that received TAI-84 90 and 96 h after MAP sponge removal. Buffaloes in the GnRH-TAI-84 group had lower estradiol concentrations at 90, 96, 102, and 108 h than those in the TAI-84 group. Ovulation in GnRH-TAI-84 buffaloes occurred 11 h earlier than that in buffaloes from the TAI-84 group. A shorter interval between AI and ovulation in GnRH-TAI-84 buffaloes (14 h vs. 25 h) led to greater pregnancies per AI (62% vs. 17%) compared to buffaloes from the TAI-84 group.

Strategies for optimal management of pediatric acute agitation in emergency settings.

Acute agitation in youth is a challenging presentation to the emergency department. In many cases, however, youth can be behaviorally de-escalated using a combination of environmental modification and verbal de-escalation. In cases where additional strategies such as pharmacologic de-escalation or physical restraint are needed, using the least restrictive means possible, including the youth in the decision-making process, and providing options are important. This paper reviews specific considerations on the approach to a youth with acute agitation and strategies and techniques to successfully de-escalate agitated youth who pose a danger to themselves and/or others.

Generalized and novel iterative scheme for best approximate solution of large and sparse augmented linear systems.

This article presents a generalized novel iterative technique for obtaining the approximate solution of systems of linear equations ( A B B T 0 ) ( x y ) = ( p q ) . Parameters involved in the scheme play a vital role to obtain the rapid and best approximate solution. The technique is applied to some important problems and the results are presented in detail. The article compares the effectiveness of the new presented iterative scheme with modified SOR-like method, generalized SOR and SOR-like method for solving large systems of linear equations. The convergence criteria for the iterative scheme are also analysed and it can be observed in the article that the theoretical derivation is supported by numerical calculations and graphical exhibition. Overall, the article provides a new and effective method with a detailed analysis of its performance and convergence criteria for solving systems of linear equations of the form presented. The newly derived technique has potential applications in various fields of science and engineering where systems of linear equations arise and need to be solved.

Carbon-Based 3D Architectures as Anodes for Lithium-Ion Battery Systems.

Graphite, with its exceptional cyclic performance, continues to dominate as the preferred anode material for lithium-ion batteries. However as high-energy application gains momentum, there is growing demand for higher capacities that alloying/de alloying and conversion type anode materials can offer. Despite their potential, these materials are plagued by challenges such as volumetric fluctuations, low conductivities, and poor cyclic stability. Carbon nanostructures, on the other hand, show tremendous promise with their low volume expansion, high ion diffusion rates, and excellent conductivity. Nevertheless, their limited areal and volumetric densities restrict their widespread utilization. To address these limitations, various strategies such as doping, composite formation, and structural modification have been proposed. This article provides a succinct overview of carbon nanomaterials and their electrochemical performance as 3D carbon-based anodes, along with a comprehensive analysis of the strategies employed to overcome associated challenges while evaluating their potential prospects in the field.

Isolation and characterization of chromium-resistant bacteria and their effects on germination, growth, and Cr accumulation in Capsicum annum (L.) under Cr stress.

Chromium (Cr) is a well-known environmental pollutant while less information is available on the role of Cr-resistant bacteria in the alleviation of Cr-stress in chili (Capsicum annum L.) plants. Effect of Cr-resistant bacterial strains on growth and Cr uptake by chili plants was investigated. The results revealed that Cr-stress showed a negative effect on germination, photosynthesis, and relative water content but the inoculation ameliorated the plant stress. Chromium-resistant bacterial strains enhanced the shoot and root growth (33% SL, 19.7% RL), shoot and root dry weight (35%, 32.9%), relative water content (32.25%), membrane stability index (46.52%) SPAD value (50.76%), Cr concentration in shoots and roots (19.87 and 18.52 mg kg-1), bioaccumulation and translocation factor (0.396 mgkg-1), and seedling vigor index (40.8%) of plants. Chromium-resistant bacterial strains enhanced the NPK uptake while reduced Cr uptake by plants. The morphological and biochemical examination of rhizobacterial strains (and NM28) resistant to Cr-stress revealed smooth, off-white colonies of bacteria composed of rod-shaped cells which are Gram positive in reaction while negative in catalase activity. High quantities of malic acid were produced by bacterial strains under study i.e. NM8 (926.12 µgmL-2) and NM28 (992.25 µgmL-2). These strains were identified as Bacillus cereus strain NM8 and Bacillus subtilis strain NM28 through 16S rRNA sequencing. Results showed that B. cereus strain NM28 is more effective than B. cereus strain NM8 in promoting the growth of Cr-stressed Chili that might be suitable to develop biofertilizer for sustainable production of vegetables under metal stress.

Soy protein-polyphenols conjugates interaction mechanism, characterization, techno-functional and biological properties: An updated review.

Soy protein is a promising nutritional source with improved functionality and bioactivities due to conjugation with polyphenols (PP)-the conjugates between soy protein and PP held by covalent and noncovalent bonds. Different approaches, including thermodynamics, spectroscopy, and molecular docking simulations, can demonstrate the outcomes and mechanism of these conjugates. The soy protein, PP structure, matrix properties (temperature, pH), and interaction mechanism alter the ζ-potential, secondary structure, thermal stability, and surface hydrophobicity of proteins and also improve the techno-functional properties such as gelling ability, solubility, emulsifying, and foaming properties. Soy protein-PP conjugates also reveal enhanced in vitro digestibility, anti-allergic, antioxidant, anticancer, anti-inflammatory, and antimicrobial activities. Thus, these conjugates may be employed as edible film additives, antioxidant emulsifiers, hydrogels, and nanoparticles in the food industry. Future research is needed to specify the structure-function associations of soy protein-PP conjugates that may affect their functionality and application in the food industry.

Food authentication, current issues, analytical techniques, and future challenges: A comprehensive review.

Food authentication and contamination are significant concerns, especially for consumers with unique nutritional, cultural, lifestyle, and religious needs. Food authenticity involves identifying food contamination for many purposes, such as adherence to religious beliefs, safeguarding health, and consuming sanitary and organic food products. This review article examines the issues related to food authentication and food fraud in recent periods. Furthermore, the development and innovations in analytical techniques employed to authenticate various food products are comprehensively focused. Food products derived from animals are susceptible to deceptive practices, which can undermine customer confidence and pose potential health hazards due to the transmission of diseases from animals to humans. Therefore, it is necessary to employ suitable and robust analytical techniques for complex and high-risk animal-derived goods, in which molecular biomarker-based (genomics, proteomics, and metabolomics) techniques are covered. Various analytical methods have been employed to ascertain the geographical provenance of food items that exhibit rapid response times, low cost, nondestructiveness, and condensability.

Prediction of Nili-Ravi buffalo bull fertility through Fourier harmonic analysis of sperm.

Fourier harmonic analysis (FHA) is a robust method for identification of minute changes in sperm nuclear shape that are indicative of reduced fertility. The current study was designed to develop a fertility prediction model for Nili-Ravi buffalo bulls through FHA of sperm. In experiment I, FHA technique was standardized, average sperm nuclear perimeter was measured and sperm nuclear shape plot of buffalo bull was constructed. Sperm of buffalo bulls (n = 10) were stained with YOYO-1 and Hoechst-33342 to differentiate live and dead, and digital images were captured using phase contrast and fluorescent microscopy. The images were analyzed by ImageJ software and 100 sperm/bull were evaluated. The results are described as mean ± SEM values of mean harmonic amplitude (mharm), skewness harmonic amplitude (skharm), kurtosis harmonic amplitude (kurharm) and variance harmonic amplitude (varharm) at Fourier frequencies 0-5 along with the cartesian and polar coordinate plots of buffalo bull sperm. In experiment II, a fertility prediction model was developed based on FHA of buffalo bull sperm. Semen samples of low (n = 6), medium (n = 3) and high (n = 8) fertility bulls were investigated for FHA of sperm and harmonic amplitudes (HA) were generated. Firstly, to determine if live and dead sperm population have unique nuclear shape distribution; the mean, skewness, kurtosis and variance HA 0-5 of 1700 live and 1294 dead spermatozoa of 17 bulls were evaluated. T-test signified a difference in the mharm0 (2.363 ± 0.01 vs. 2.439 ± 0.02), skharm0 (-0.0002 ± 0.07 vs. -0.266 ± 0.09), kurharm0 (-0.156 ± 0.07 vs. 0.260 ± 0.18), kurharm2 (0.142 ± 0.11 vs. 1.031 ± 0.32) and varharm4 (0.109 ± 0.00 vs. 0.082 ± 0.00) of live vs. dead sperm population (p < 0.05). Therefore, 100 live sperm/bull were further evaluated for mean, skewness, kurtosis and variance HA 0-5 values among high (n = 6) and low-fertility (n = 6) groups. Results of T-test showed higher values of mharm2 (0.739 ± 0.01 vs. 0.686 ± 0.00), mharm4 (0.105 ± 0.001 vs. 0.007 ± 0.001), and skharm0 (0.214 ± 0.109 vs. -0.244 ± 0.097) in high vs. low-fertility group (p < 0.05). In next step, five significantly different combinations of discriminant measures between high and low-fertility groups were obtained by discriminant analysis. In conclusion, mharm4, skharm0 and varharm2 correctly identified 91.7 % of bulls into their respective fertility groups, and upon cross validation the value of the canonical correlation was 0.928.

Chronic cavitary pulmonary aspergillosis as a sequela of pulmonary tuberculosis: A case report from Pakistan.

Chronic pulmonary aspergillosis is a lung disorder characterized by the presence of single or multiple cavities with or without an aspergilloma or nodules on chest imaging, with mycological evidence of and/or demonstration of immunological response to Aspergillus spp. The affected patient should manifest relevant symptoms for at least 3 months. Chronic cavitary pulmonary aspergillosis is the most common subset of chronic pulmonary aspergillosis, which is often reported in patients previously treated for pulmonary tuberculosis, having residual cavities in their lungs. We present a case of a 55-year-old male patient treated for pulmonary tuberculosis 2 years back, now presenting with fever, shortness of breath, and hemoptysis with overt radiological changes from the baseline, positive direct microscopy, and serology for Aspergillus spp. and thus meeting the criteria for chronic cavitary pulmonary aspergillosis. Treatment with oral antifungal was initiated, but the follow-up data are unavailable due to patient noncompliance and lack of resources. We aim to emphasize the radiological and microbiological features of this condition to aid the early diagnosis and prompt treatment, as this may mimic similar pulmonary disorders and pose a significant challenge in the diagnosis and management outcomes.

Mycotoxin patulin contamination in various fruits and estimating its dietary impact on the consumers: From orchard to table.

The present research examined patulin's presence across the whole supply chain of selected fruits. A comprehensive analysis was conducted on 442 samples of fruits (oranges, apples, apricots, lemons, and guava) to determine the presence of patulin contamination. This analysis used Liquid Chromatography (HPLC) with a UV detector. The findings indicate that 17, 23, and 28 % of selected fruit samples tested positive for patulin levels in farm, transportation, and market samples. However, the sample collected during the transportation step showed that 56 % (percentage of positive samples) of fruits have patulin levels greater than 50 µg/kg, and 41 % (percentage of positive samples) have greater levels than 50 µg/kg in market samples. The findings of the one-way analysis of variance indicated that no statistically significant variation existed between the amounts of patulin across the various stages of the food supply chain system (p > 0.05). Nevertheless, the analysis of the correlation study, namely Kendall's tau_b and Spearman's rho, denote a robust association between the levels of patulin and the food supply system. The apple samples exhibited the most significant average dietary intake of patulin, with an average value of 0.11 µg/kg bw/day. The maximum mean hazard quotient (HQ) of 0.28 was also recorded. The prevalence and incidence of patulin in specific fruits were found to be relatively high, and it was observed that market samples had elevated levels of patulin in the selected fruits.

Correlation Between Body Mass Index and Apnea-Hypopnea Index or Nadir Oxygen Saturation Levels in Patients With Obstructive Sleep Apnea.

BACKGROUND: Apnea-hypopnea index (AHI) and nadir oxygen saturation (SpO2) are the indexes used to measure the severity of obstructive sleep apnea (OSA). Obesity, measured by body mass index (BMI), is one of the main contributing factors to the onset and severity of OSA in patients. This study was conducted to find the association between BMI and OSA severity indexes, mainly AHI and nadir SpO2 levels. METHODS: Polysomnography reports of patients with diagnosed OSA in a teaching hospital were retrospectively reviewed. BMI, AHI, and nadir SpO2 levels were recorded from the sleep study reports of the patients. Spearman's Rho test was applied to find the correlation between BMI and AHI/nadir Spo2 levels. RESULTS: A total of 167 patients were included in the study, comprising 83 males and 84 females. The Mann-Whitney U test was utilized to investigate the association between BMI and gender and age groups. The analysis revealed a significant difference in BMI between males and females, with females having a higher BMI. However, there was no significant difference in BMI among individuals in the early middle and late middle age groups. Spearman's Rho test was employed to explore the correlation between BMI and AHI/nadir SpO2 levels. The results indicated no significant correlation between BMI and AHI (p = .122) or nadir SpO2 levels (p = .239). CONCLUSION: Contrary to common belief, BMI was not linked to the severity of OSA. It implies that several other factors, independent of BMI, play a role in the disease progression and severity.

Molecular identification of Proteus mirabilis, Vibrio species leading to CRISPR-Cas9 modification of tcpA and UreC genes causing cholera and UTI.

Heavy metal accumulation increases rapidly in the environment due to anthropogenic activities and industrialization. The leather and surgical industry produces many contaminants containing heavy metals. Cadmium, a prominent contaminant, is linked to severe health risks, notably kidney and liver damage, especially among individuals exposed to contaminated wastewater. This study aims to leverage the natural cadmium resistance mechanisms in bacteria for bioaccumulation purposes. The industrial wastewater samples, characterized by an alarming cadmium concentration of 29.6 ppm, 52 ppm, and 76.4 ppm-far exceeding the recommended limit of 0.003 ppm-were subjected to screening for cadmium-resistant bacteria using cadmium-supplemented media with CdCl2. 16S rRNA characterization identified Vibrio cholerae and Proteus mirabilis as cadmium-resistant bacteria in the collected samples. Subsequently, the cadmium resistance-associated cadA gene was successfully amplified in Vibrio species and Proteus mirabilis, revealing a product size of 623 bp. Further analysis of the identified bacteria included the examination of virulent genes, specifically the tcpA gene (472 bp) associated with cholera and the UreC gene (317 bp) linked to urinary tract infections. To enhance the bioaccumulation of cadmium, the study proposes the potential suppression of virulent gene expression through in-silico gene-editing tools such as CRISPR-Cas9. A total of 27 gRNAs were generated for UreC, with five selected for expression. Similarly, 42 gRNA sequences were generated for tcpA, with eight chosen for expression analysis. The selected gRNAs were integrated into the lentiCRISPR v2 expression vector. This strategic approach aims to facilitate precise gene editing of disease-causing genes (tcpA and UreC) within the bacterial genome. In conclusion, this study underscores the potential utility of Vibrio species and Proteus mirabilis as effective candidates for the removal of cadmium from industrial wastewater, offering insights for future environmental remediation strategies.

Viability of Free and Alginate-Carrageenan Gum Coated Lactobacillus acidophilus and Lacticaseibacillus casei in Functional Cottage Cheese.

The survivability of encapsulated and nonencapsulated probiotics consisting of Lactobacillus acidophilus and Lacticaseibacillus casei and the nutritional, physicochemical, and sensorial features of cottage cheese were investigated under refrigeration storage at 4 °C for 28 days. Microbeads of L. acidophilus and L. casei were developed using 2% sodium alginate, 1.5% sodium alginate and 0.5% carrageenan, and 1% sodium alginate and 1% carrageenan using an encapsulation technique to assess the probiotic viability in cottage cheese under different gastrointestinal conditions (SGF (simulated gastric juice), SIF (simulated intestinal fluid)), and bile salt) and storage conditions. Scanning electron microscopy (SEM) elucidated the stable structure of microbeads, Fourier transform infrared spectroscopy (FTIR) confirmed the presence probiotics in the microcapsules, and X-ray diffraction (XRD) demonstrated the amorphous state of microbeads. Furthermore, the highest encapsulation efficiency was observed for alginate 1% and carrageenan 1% microbeads (T3), i.e., 95%. Likewise, viability was recorded in T3 against SGF, SIF, and bile salt solution, i.e., 8.5, 8.8, and 8.9 log CFU/g at 80 min of exposure, compared to the control. The results of pH showed a significant (p < 0.05) decline that ultimately increased the titratable acidity. Nutritional analysis of cottage cheese revealed the highest levels of ash, protein, and total solids in T3, exhibiting mean values of 3.2, 22, and 43.2 g/100 g, respectively, after 28 days of storage. The sensory evaluation of cottage cheese demonstrated better color, flavor, and textural attributes in T3. Conclusively, synergistic addition of L. acidophilus and L. casei encapsulated with alginate-carrageenan gums was found to be more effective in improving the viability of probiotics in cottage cheese than noncapsulated cells while carrying better magnitudes of ash and protein, lower acidity, and pleasant taste.