Analyzing Antibiotic Resistance in Bacteria from Wastewater in Pakistan Using Whole-Genome Sequencing.

Background: Wastewater is a major source of Antibiotic-Resistant Bacteria (ARB) and a hotspot for the exchange of Antibiotic-Resistant Genes (ARGs). The occurrence of Carbapenem-Resistant Bacteria (CRB) in wastewater samples is a major public health concern. Objectives: This study aimed to analyze Antibiotic resistance in bacteria from wastewater sources in Pakistan. Methods: We analyzed 32 bacterial isolates, including 18 Escherichia coli, 4 Klebsiella pneumoniae, and 10 other bacterial isolates using phenotypic antibiotic susceptibility assay and whole-genome sequencing. This study identified the ARGs, plasmid replicons, and integron genes cassettes in the sequenced isolates. One representative isolate was further sequenced using Illumina and Oxford nanopore sequencing technologies. Results: Our findings revealed high resistance to clinically important antibiotics: 91% of isolates were resistant to cefotaxime, 75% to ciprofloxacin, and 62.5% to imipenem, while 31% showed non-susceptibility to gentamicin. All E. coli isolates were resistant to cephalosporins, with 72% also resistant to carbapenems. Sequence analysis showed a diverse resistome, including carbapenamases (blaNDM-5, blaOXA-181), ESBLs (blaCTX-M-15, blaTEM), and AmpC-type ß-lactamases (blaCMY). Key point mutations noticed in the isolates were pmrB_Y358N (colistin) and ftsI_N337NYRIN, ftsI_I336IKYRI (carbapenem). The E. coli isolates had 11 different STs, with ST410 predominating (28%). Notably, the E. coli phylogroup A isolate 45EC1, (ST10886) is reported for the first time from wastewater, carrying blaNDM-5, blaCMY-16, and pmrB_Y358N with class 1 integron gene cassette of dfrA12-aadA2-qacEΔ1 on a plasmid-borne contig. Other carbapenamase, blaNDM-1 and blaOXA-72, were detected in K. pneumoniae 22EB1 and Acinetobacter baumannii 51AC1, respectively. The integrons with the gene cassettes encoding antibiotic resistance, and the transport and bacterial mobilization protein, were identified in the sequenced isolates. Ten plasmid replicons were identified, with IncFIB prevalent in 53% of isolates. Combined Illumina and Oxford nanopore sequencing revealed blaNDM-5 on an IncFIA/IncFIC plasmid and is identical to those reported in the USA, Myanmar, and Tanzania. Conclusions: These findings highlight the environmental prevalence of high-risk and WHO-priority pathogens with clinically important ARGs, underscoring the need for a One Health approach to mitigate ARB isolates.

Exploring Lignin Biosynthesis Genes in Rice: Evolution, Function, and Expression.

Lignin is nature's second most abundant vascular plant biopolymer, playing significant roles in mechanical support, water transport, and stress responses. This study identified 90 lignin biosynthesis genes in rice based on phylogeny and motif constitution, and they belong to PAL, C4H, 4CL, HCT, C3H, CCoAOMT, CCR, F5H, COMT, and CAD families. Duplication events contributed largely to the expansion of these gene families, such as PAL, CCoAOMT, CCR, and CAD families, mainly attributed to tandem and segmental duplication. Microarray data of 33 tissue samples covering the entire life cycle of rice suggested fairly high PAL, HCT, C3H, CCoAOMT, CCR, COMT, and CAD gene expressions and rather variable C4H, 4CL, and F5H expressions. Some members of lignin-related genes (OsCCRL11, OsHCT1/2/5, OsCCoAOMT1/3/5, OsCOMT, OsC3H, OsCAD2, and OsPAL1/6) were expressed in all tissues examined. The expression patterns of lignin-related genes can be divided into two major groups with eight subgroups, each showing a distinct co-expression in tissues representing typically primary and secondary cell wall constitutions. Some lignin-related genes were strongly co-expressed in tissues typical of secondary cell walls. Combined HPLC analysis showed increased lignin monomer (H, G, and S) contents from young to old growth stages in five genotypes. Based on 90 genes' microarray data, 27 genes were selected for qRT-PCR gene expression analysis. Four genes (OsPAL9, OsCAD8C, OsCCR8, and OsCOMTL4) were significantly negatively correlated with lignin monomers. Furthermore, eleven genes were co-expressed in certain genotypes during secondary growth stages. Among them, six genes (OsC3H, OsCAD2, OsCCR2, OsCOMT, OsPAL2, and OsPAL8) were overlapped with microarray gene expressions, highlighting their importance in lignin biosynthesis.

Molasses-based waste water irrigation: a friend or foe for carrot (Daucus carota L.) growth, yield and nutritional quality.

Management of molasses-based wastewater generated in yeast and sugar industries is a major environmental concern due to its high chemical oxygen demand and other recalcitrant substances. Several strategies have been used to reduce the inland discharge of wastewater but the results are not satisfactory due to high operating cost. However, reuse of molasses-based wastewater irrigation in agriculture has been a major interest nowadays to reduce the freshwater consumption. Thus, it is crucial to monitor the impacts of molasses-based waste water irrigation on growth, metabolism, yield and nutritional quality of crops for safer consumer's health. In present study, carrot seeds of a local cultivar (T-29) were germinated on filter paper in Petri dishes under controlled conditions. The germinated seeds were then transplanted into pots and irrigated with three different treatments normal water (T0), diluted molasses-based wastewater (T1), and untreated molasses-based wastewater (T2), in six replicates. Results revealed that carrot irrigated with untreated molasses-based waste water had exhibited significant reductions in growth, yield, physiology, metabolism, and nutritional contents. Additionally, accumulation of Cd and Pb contents in carrot roots irrigated with untreated molasses-based waste water exceed the permissible limits suggested by WHO and their consumption may cause health risks. While, diluted molasses-based waste water irrigation positively enhanced the growth, yield of carrot plants without affecting the nutritional quality. This strategy is cost effective, appeared as most appropriate alternative mean to reduce the freshwater consumption in water deficit regions of the world.

To PE or Not to PE: Perfusion SPECT/CT Avoids Overdiagnosis.

ABSTRACT: A 41-year-old woman with metastatic breast cancer presented with dyspnea, hypoxia, and elevated d -dimer. Perfusion planar imaging followed by SPECT/CT of the chest was performed due to the patient's iodinated contrast allergy. Planar images showed multiple pleural-based wedge-shaped defects concerning for bilateral pulmonary embolism (PE). Perfusion SPECT/CT of the chest confirmed multiple areas of perfusion defects but was considered negative for PE and attributed the perfusion defects to the compressing of pulmonary vasculature from metastatic lymph nodes and pulmonary masses. Given the high pretest probability of PE, a CT pulmonary angiogram was performed after premedication for contrast allergy confirming absence of PE.

Thrombocytopenia and Grading of Esophageal Varices in Patients With Chronic Liver Disease.

Background Chronic liver disease (CLD) is associated with a variety of consequences, including thrombocytopenia and esophageal varices, which significantly impact patient prognosis and management. Thrombocytopenia, frequently observed in patients with CLD, may correlate with the severity of esophageal varices, a critical complication leading to variceal bleeding. Methodology A cross-sectional study was carried out in the Department of Medicine and Gastroenterology, Pak Emirates Military Hospital, Rawalpindi, from October 2021 to March 2022. The study enrolled 94 patients, aged 18-70 years, diagnosed with CLD, regardless of the cause. These patients were categorized into four groups based on platelet count: <50,000/uL, 50,000-99,999/uL, 100,000-150,000/uL, and >150,000/uL. Pearson's correlation was utilized to evaluate the association between the severity of thrombocytopenia and the grading of esophageal varices. Results A total of 94 patients were enrolled in the study, with 53 (56.4%) males and 41 (43.6%) females. The mean age of patients was 51.06 ±11.09 years. Seventeen (18.1%) had no esophageal varices, 16 (17.0%) were diagnosed with Grade I varices, 35 (37.2%) with Grade II varices, and 26 (27.7%) had Grade III varices. Most patients without varices had a platelet count above 150 x 103 (17, 18.1%). Conversely, most patients with Grade III varices (19, 20.2%) had platelet counts below 50 x 103. Patients with no esophageal varices had a mean platelet count of 173.70 ± 37.48 x 103. Among the patients, those with Grade III esophageal varices exhibited the lowest mean platelet count, recorded at 78.54 ± 24.14 x 103. These findings indicate a statistically significant difference in mean platelet counts across the various esophageal varices grades (P = 0.000). There was an inverse correlation of platelet count with the grading of esophageal varices (r = -0.645, P < 0.000). Conclusions A negative correlation was observed between the platelet count and the grading of esophageal varices, implying that as the severity of esophageal varices increased, the platelet counts proportionally decreased.

Green extraction and application of yellow natural curcumin colorant from Curcuma aromatica rhizomes for silk dyeing.

Green products such as plant tints are becoming more and more well-known worldwide due to their superior biological and ayurvedic properties. In this work, colorant from Amba Haldi (Curcuma aromatica) was isolated using microwave (MW), and bio-mordants were added to produce colorfast shades. Response surface methodology was used to develop a central composite design (CCD), which maximizes coloring variables statistically. The findings from 32 series of experiments show that excellent color depth (K/S = 12.595) was established onto MW-treated silk fabric (RS = 4 min) by employing 65 mL of radiated aqueous extract (RE = 4 min) of 5 pH cutting-edge the existence of 1.5 g/100 mL used sodium chloride at 75 °C for 45 min. It was discovered that acacia (keekar) extract (1%), pomegranate extract (2%), and pistachio extract (1.5%) were present before coloring by the use of bio-mordants. On the other hand, upon dyeing, acacia extract (1.5%), pomegranate extract (1.5%), and pistachio extract (2%) have all shown extremely strong colorfast colors. Comparatively, before dyeing, salts of Al3+ (1.5%), Fe2+ (2%), and TA (1.5%) gave good results; after dyeing, salts of Al3+ (1%) and Fe2+ (1.5%) and TA (2%) gave good results. When applied to silk fabric, MW radiation has increased the production of dyes recovered from rhizomes. Additionally, the right amount of chemical and biological mordants have been added, resulting in color fastness ratings ranging from outstanding to good. Therefore, the natural color extracted from Amba Haldi can be a sustainable option for the dyeing of silk fabric in the textile dyeing and finishing industries.

Characterization of clumpy adhesion of Escherichia coli to human cells and associated factors influencing antibiotic sensitivity.

Escherichia coli intestinal infection pathotypes are characterized by distinct adhesion patterns, including the recently described clumpy adhesion phenotype. Here, we identify and characterize the genetic factors contributing to the clumpy adhesion of E. coli strain 4972. In this strain, the transcriptome and proteome of adhered bacteria were found to be distinct from planktonic bacteria in the supernatant. A total of 622 genes in the transcriptome were differentially expressed in bacteria present in clumps relative to the planktonic bacteria. Seven genes targeted for disruption had variable distribution in different pathotypes and nonpathogenic E. coli, with the pilV and spnT genes being the least frequent or absent from most groups. Deletion (Δ) of five differentially expressed genes, flgH, ffp, pilV, spnT, and yggT, affected motility, adhesion, or antibiotic stress. ΔflgH exhibited 80% decrease and ΔyggT depicted 184% increase in adhesion, and upon complementation, adhesion was significantly reduced to 13%. ΔflgH lost motility and was regenerated when complemented, whereas Δffp had significantly increased motility, and reintroduction of the same gene reduced it to the wild-type level. The clumps produced by Δffp and ΔspnT were more resistant and protected the bacteria, with ΔspnT showing the best clump formation in terms of ampicillin stress protection. ΔyggT had the lowest tolerance to gentamicin, where the antibiotic stress completely eliminated the bacteria. Overall, we were able to investigate the influence of clump formation on cell surface adhesion and antimicrobial tolerance, with the contribution of several factors crucial to clump formation on susceptibility to the selected antibiotics. IMPORTANCE: The study explores a biofilm-like clumpy adhesion phenotype in Escherichia coli, along with various factors and implications for antibiotic susceptibility. The phenotype permitted the bacteria to survive the onslaught of high antibiotic concentrations. Profiles of the transcriptome and proteome allowed the differentiation between adhered bacteria in clumps and planktonic bacteria in the supernatant. The deletion mutants of genes differentially expressed between adhered and planktonic bacteria, i.e., flgH, ffp, pilV, spnT, and yggT, and respective complementations in trans cemented their roles in multiple capacities. ffp, an uncharacterized gene, is involved in motility and resistance to ampicillin in a clumpy state. The work also affirms for the first time the role of the yggT gene in adhesion and its involvement in susceptibility against another aminoglycoside antibiotic, i.e., gentamicin. Overall, the study contributes to the mechanisms of biofilm-like adhesion phenotype and understanding of the antimicrobial therapy failures and infections of E. coli.

Structural basis for the phase separation of the chromosome passenger complex.

The physical basis of phase separation is thought to consist of the same types of bonds that specify conventional macromolecular interactions yet is unsatisfyingly often referred to as 'fuzzy'. Gaining clarity on the biogenesis of membraneless cellular compartments is one of the most demanding challenges in biology. Here, we focus on the chromosome passenger complex (CPC), that forms a chromatin body that regulates chromosome segregation in mitosis. Within the three regulatory subunits of the CPC implicated in phase separation - a heterotrimer of INCENP, Survivin, and Borealin - we identify the contact regions formed upon droplet formation using hydrogen/deuterium exchange mass spectrometry (HXMS). These contact regions correspond to some of the interfaces seen between individual heterotrimers within the crystal lattice they form. A major contribution comes from specific electrostatic interactions that can be broken and reversed through initial and compensatory mutagenesis, respectively. Our findings reveal structural insight for interactions driving liquid-liquid demixing of the CPC. Moreover, we establish HXMS as an approach to define the structural basis for phase separation.

A novel solution to optimal power flow problems using composite differential evolution integrating effective constrained handling techniques.

Optimal power flow is a complex and highly non-linear problem in which steady-state parameters are needed to find a network's efficient and economical operation. In addition, the difficulty of the Optimal power flow problem becomes enlarged when new constraints are added, and it is also a challenging task for the power system operator to solve the constrained Optimal power flow problems efficiently. Therefore, this paper presents a constrained composite differential evolution optimization algorithm to search for the optimum solution to Optimal power flow problems. In the last few decades, numerous evolutionary algorithm implementations have emerged due to their superiority in solving Optimal power flow problems while considering various objectives such as cost, emission, power loss, etc. evolutionary algorithms effectively explore the solution space unconstrainedly, often employing the static penalty function approach to address the constraints and find solutions for constrained Optimal power flow problems. It is a drawback that combining evolutionary algorithms and the penalty function approach requires several penalty parameters to search the feasible space and discard the infeasible solutions. The proposed a constrained composite differential evolution algorithm combines two effective constraint handling techniques, such as feasibility rule and ɛ constraint methods, to search in the feasible space. The proposed approaches are recognized on IEEE 30, 57, and 118-bus standard test systems considering 16 study events of single and multi-objective optimization functions. Ultimately, simulation results are examined and compared with the many recently published techniques of Optimal power flow solutions owing to show the usefulness and performance of the proposed a constrained composite differential evolution algorithm.

Exploring water-absorbing capacity: a digital image analysis of seeds from 120 wheat varieties.

Wheat is a staple food crop that provides a significant portion of the world's daily caloric intake, serving as a vital source of carbohydrates and dietary fiber for billions of people. Seed shape studies of wheat typically involve the use of digital image analysis software to quantify various seed shape parameters such as length, width, area, aspect ratio, roundness, and symmetry. This study presents a comprehensive investigation into the water-absorbing capacity of seeds from 120 distinct wheat lines, leveraging digital image analysis techniques facilitated by SmartGrain software. Water absorption is a pivotal process in the early stages of seed germination, directly influencing plant growth and crop yield. SmartGrain, a powerful image analysis tool, was employed to extract precise quantitative data from digital images of wheat seeds, enabling the assessment of various seed traits in relation to their water-absorbing capacity. The analysis revealed significant transformations in seed characteristics as they absorbed water, including changes in size, weight, shape, and more. Through statistical analysis and correlation assessments, we identified robust relationships between these seed traits, both before and after water treatment. Principal Component Analysis (PCA) and Agglomerative Hierarchical Clustering (AHC) were employed to categorize genotypes with similar trait patterns, providing insights valuable for crop breeding and genetic research. Multiple linear regression analysis further elucidated the influence of specific seed traits, such as weight, width, and distance, on water-absorbing capacity. Our study contributes to a deeper understanding of seed development, imbibition, and the crucial role of water absorption in wheat. These insights have practical implications in agriculture, offering opportunities to optimize breeding programs for improved water absorption in wheat genotypes. The integration of SmartGrain software with advanced statistical methods enhances the reliability and significance of our findings, paving the way for more efficient and resilient wheat crop production. Significant changes in wheat seed shape parameters were observed after imbibition, with notable increases in area, perimeter, length, width, and weight. The length-to-width ratio (LWR) and circularity displayed opposite trends, with higher values before imbibition and lower values after imbibition.

Punicalagin, a pomegranate polyphenol sensitizes the activity of antibiotics against three MDR pathogens of the Enterobacteriaceae.

BACKGROUND: Multidrug resistance (MDR) in the family Enterobacteriaceae is a perniciously increasing threat to global health security. The discovery of new antimicrobials having the reversing drug resistance potential may contribute to augment and revive the antibiotic arsenal in hand. This study aimed to explore the anti-Enterobacteriaceae capability of bioactive polyphenols from Punica granatum (P. granatum) and their co-action with antibiotics against clinical isolates of Enterobacteriaceae predominantly prevalent in South Asian countries. METHODS: The Kandhari P. granatum (Pakistani origin) extracts were tested for anti-Enterobacteriaceae activity by agar well diffusion assay against MDR Salmonella enterica serovar Typhi, serovar Typhimurium and Escherichia coli. Predominant compounds of active extract were determined by mass spectrometry and screened for bioactivity by agar well diffusion and minimum inhibitory concentration (MIC) assay. The active punicalagin was further evaluated at sub-inhibitory concentrations (SICs) for coactivity with nine conventional antimicrobials using a disc diffusion assay followed by time-kill experiments that proceeded with SICs of punicalagin and antimicrobials. RESULTS: Among all P. granatum crude extracts, pomegranate peel methanol extract showed the largest inhibition zones of 25, 22 and 19 mm, and the MICs as 3.9, 7.8 and 7.8 mg/mL for S. typhi, S. typhimurium and E. coli, respectively. Punicalagin and ellagic acid were determined as predominant compounds by mass spectrometry. In plate assay, punicalagin (10 mg/mL) was active with hazy inhibition zones of 17, 14, and 13 mm against S. typhi, S. typhimurium and E. coli, respectively. However, in broth dilution assay punicalagin showed no MIC up to 10 mg/mL. The SICs 30 µg, 100 µg, and 500 µg of punicalagin combined with antimicrobials i.e., aminoglycoside, ß-lactam, and fluoroquinolone act in synergy against MDR strains with % increase in inhibition zone values varying from 3.4 ± 2.7% to 73.8 ± 8.4%. In time-kill curves, a significant decrease in cell density was observed with the SICs of antimicrobials/punicalagin (0.03-60 µg/mL/30, 100, 500 µg/mL of punicalagin) combinations. CONCLUSIONS: The P. granatum peel methanol extract exhibited antimicrobial activity against MDR Enterobacteriaceae pathogens. Punicalagin, the bacteriostatic flavonoid act as a concentration-dependent sensitizing agent for antimicrobials against Enterobacteriaceae. Our findings for the therapeutic punicalagin-antimicrobial combination prompt further evaluation of punicalagin as a potent activator for drugs, which otherwise remain less or inactive against MDR strains.

Chromosomal passenger complex condensates generate parallel microtubule bundles in vitro.

The mitotic spindle contains many bundles of microtubules (MTs) including midzones and kinetochore fibers, but little is known about how bundled structures are formed. Here, we show that the chromosomal passenger complex (CPC) purified from Escherichia coli undergoes liquid-liquid demixing in vitro. An emergent property of the resultant condensates is to generate parallel MT bundles when incubated with free tubulin and GTP in vitro. We demonstrate that MT bundles emerge from CPC droplets with protruding minus ends that then grow into long and tapered MT structures. During this growth, we found that the CPC in these condensates apparently reorganize to coat and bundle the resulting MT structures. CPC mutants attenuated for liquid-liquid demixing or MT binding prevented the generation of parallel MT bundles in vitro and reduced the number of MTs present at spindle midzones in HeLa cells. Our data demonstrate that an in vitro biochemical activity to produce MT bundles emerges after the concentration of the CPC and provides models for how cells generate parallel-bundled MT structures that are important for the assembly of the mitotic spindle. Moreover, these data suggest that cells contain MT-organizing centers that generate MT bundles that emerge with the opposite polarity from centrosomes.

Characterization of Salmonella enterica Biofilms and Antibiofilm Effect of Carvacrol and 2-Aminobenzimidazole.

Biofilm-associated foodborne Salmonella infections in poultry have become increasingly challenging for veterinarians, particularly in developing countries, and warrant thorough investigation. We assessed the biofilm-forming tendency of poultry isolates of Salmonella enterica, namely Salmonella Typhimurium (n = 23), Salmonella Infantis (n = 28), and Salmonella Heidelberg (n = 18), in nutrient-rich Rappaport-Vassiliadis Soya (RVS) peptone broth and nutrient-deficient diluted Tryptone Soya Broth (TSB). Seven of the tested isolates exhibited moderate biofilm formation in diluted TSB, whereas two showed such formation in RVS. In addition, the Congo red agar assay revealed curli and cellulose production in seven isolates. Fourteen specific biofilm-associated genes were analyzed identifying sdiA and seqA to be the most prevalent (100%), and glyA the least prevalent (69.5%). The prevalence of the genes bcsA and csgA was significantly lower in moderate and weak biofilm formers, respectively, as compared with nonbiofilm formers in RVS peptone broth. Furthermore, the compounds carvacrol and 2-aminobenzimidazole (2-ABI) effectively inhibited biofilm formation by Salmonella serovars in RVS peptone and TSB media, respectively. Whereas the antibiofilm activity of 2-ABI against Salmonella has not been reported previously, we determined its most effective concentration at 1.5 mM among tested antibiofilm treatments. These findings indicate that Salmonella strains prevalent in poultry farms have the potential to form biofilms, and the tested compounds should be further explored as supportive or alternative antimicrobials.

Scorpion sting in Hafizabad, Pakistan: Facility based epidemiological study.

Scorpion envenomation is a widespread issue in tropical and subtropical countries. In the present study epidemiology of scorpion sting cases and potential treatment options in district Hafizabad, Pakistan are documented. Hafizabad city and the adjacent villages were selected for the data collection. Age, gender, time, site of scorpion sting on the body, scorpion sting symptoms, number of patients obtaining medical attention and the number of fatalities (if any) were noted. Data showed that scorpion envenomation often occurs in people between the ages of 16-47 years. There were more female victims (55%) and most of the scorpion sting incidences (55.49%) occurred between 07 PM and 02 AM Among body parts, both the hands (41.71%) and feet (48.57%) were more vulnerable to scorpion stings. In rural areas, the incidences of scorpion stings were higher (68.57%). The reported symptoms of scorpion stings include pain, hypertension, nausea and allergy. Out of all the victims, only 56.05% sought treatment from health care units as well as traditional medicine practitioners. However, others (43.94%) fully healed on their own without any treatment. People were using different types of plant-based materials and dead scorpions with mustard oil as a potent remedy against scorpion stings. Moreover, in the health care units there was no antivenom available and medical staff usually used common pain killers such as Xylocain and Lignocain for the treatment of scorpion sting.

Presence of fimH and iss type 1, 2 and 3 genes in uropathogenic Escherichia coli isolates recovered from an apex medical institute in North India.

PURPOSE: To detect the presence of fimH and iss type 1, 2 and 3 genes in uropathogenic Escherichia coli (UPEC) isolates recovered from patients coming to the out patient department (OPD) of our hospital. METHODS: E. coli isolates recovered from patients who had symptoms of urinary tract infection (UTI) were processed for the presence of fimH and iss genes. DNA was extracted using an in house method after which conventional PCR using forward and reverse primers targeting the four genes was carried out. The amplified products were electrophoresed and visualized in a gel documentation imager. Relevant demographic details of the patients were recorded on a pre-designed pro-forma and antimicrobial susceptibility testing of the isolates was done by disc diffusion method. RESULTS: fimH was present in 87.5% of UPEC isolates whereas iss type 1 was seen in 7.3%, type 2 in 4.2% and iss type 3 in 71.9% isolates. Age of the patients ranged from 3 months to 82 â€‹yrs (mean 43.5 SD â€‹± â€‹18.20). UTI was more common in females (60.2%) as compared to males patients (39.8%). Dysuria (66.7%) was the most common symptom in the studied subjects and diabetes mellitus (42.6%) the most common co-morbidity. A total of 56.5% patients gave a history of prior antibiotic intake. The UPEC isolates were resistant to most of the antibiotics tested. However all the isolates were sensitive to polymyxin B and colistin. Fosfomycin resistance was seen in 9.5% of the UPEC isolates harbouring fimH gene. CONCLUSION: This is the first study that highlights the presence of iss type 3 gene in UPEC isolates along with the fimH and iss type 1 and 2 genes. The results of this study can serve as a stepping stone for future in depth research into the significance of the iss genes in causing UTI.

Effect of amino acid fortified mulberry leaves on economic and biological traits of Bombyx mori L.

The demand for silk has been increasing day by day but the average silk production is not enough to meet its demand. In this study, we investigated the effect of amino acid supplemented mulberry feed on the biological and commercial traits of Bombyx mori L. (Lepidoptera; Bombycidae). The silkworm larvae at 5th instar stage were taken and fed with fresh and healthy mulberry leaves coated with Alanine, Glycine and Serine in fourteen different combinations. Results of the current study revealed that the average weight of silkworm larvae and the % ratio of silk gland to body weight on day 7 was significantly (P˂0.05) higher in the group fed with amino acid fortified leaves as compared to the control. The commercial traits of larvae fed with amino acid fortified leaves also improved significantly. The larvae fed with Alanine (1 %) treated mulberry leaves showed the maximum cocoon weight, cocoon length, cocoon width, cocoon shell ratio and fibroin content as compared to the control group. It is evident from the results that the amino acid (particularly alanine) coated mulberry leaves have a positive effect on the commercial and biological traits of Bombyx mori (L.).

Biomolecules based hydrogels and their potential biomedical applications: A comprehensive review.

The need for biocompatible drug carriers has been significantly increased from the past few years. Researchers show great interest in the development of more versatile and sophisticated biomaterials based drug carriers. Hydrogels are beneficial drug carriers and easily release the controlled amount of drug at target site due to its tunable structure. The hydrogels made-up of potent biological macromolecules including collagen, gelatin, fibrin, elastin, fibroin, chitosan, starch, alginate, agarose and carrageenan have been proven as versatile biomaterials. These are three-dimensional polymeric networks, synthesized by crosslinking of hydrophilic polymers. The biological macromolecules based hydrogels containing therapeutic substances are used in a wide range of biomedical applications including wound healing, tissue engineering, cosmetics and contact lenses. However, many aspects related to hydrogels such as the mechanism of cross-linking and molecular entanglement are not clear. So, there is a need to do more research and exploration toward the extensive and cost-effective use of hydrogels. The present review article elaborately discusses the biomolecules based hydrogels and their possible biomedical applications in different fields.

Proximate composition and prevalence and exposure assessment of aflatoxins intake through consumption of fresh carrot and processed marketed carrot products in South Punjab, Pakistan.

Carrot (Daucus carota L.) is a nutrient-rich vegetable that is widely cultivated and consumed in Pakistan in both raw and processed form. Data on the proximate composition and natural occurrence of aflatoxins (AFs) in carrots and marketed carrot products is lacking in Pakistan and the risk exposure of AF has not been characterised before. Thus, the current study was designed to know the frequently consumed carrot products with per capita consumption, and risk assessment of AF through these products in various regions of South Punjab Pakistan. A survey was conducted with 125 respondents and appeared that raw carrot, fresh carrot juice, gajrella and pickle are the most frequently consumed marketed carrot products with per capita consumption i.e. 62.5, 46.6, 16.2 and 14.5 gday-1, respectively. Proximate analysis revealed that carrot root and processed carrot products contained 9.65-98.2% moisture, 0.23-0.60% ash, 6.2-14.1% carbohydrates, 0.31-0.80% protein, 0.40-3.7% fat and 1.4-4.20% fibre. AF analysis revealed that 36.67% of samples were contaminated with TAF. Thirty-five (35%) percent of samples were tainted with aflatoxin B1, and 13.33% of samples were contaminated with aflatoxin B2. All the samples of carrot root, fresh carrot juice and gajrella contained TAF levels less than the maximum limit (ML) (2 ppb) assigned by the European Union (EU). However, the entire AFB1 positive samples of carrot pickle contained AFB1 levels of more than 2 ppb exceeding the ML. Furthermore, daily dietary exposure of TAFs ranged from 0.11 to 1.27 ng/kg of body weight per day which relatively exceeds the permissible limit of 1 ng/kg of body weight per day as defined by the Joint FAO/WHO Expert Committee on Food Additives. This is the first prevalence and risk assessment report of AF in marketed processed carrot products in Pakistan. These baseline data are an initial step in the effort to deal with this significant food safety issue and the establishment of legislation for AF in marketed products is needed in Pakistan.