Genetic Diversity and Virulence Profiling of Multi-Drug Resistant Escherichia coli of Human, Animal, and Environmental Origins.

Rapid urbanization has increased human-animal interaction and consequently enhanced the chances to acquire zoonotic diseases. The current investigation is focused to uncover the genetic diversity of multidrug-resistant E. coli strains between different ecologies (i.e., humans, livestock, and environment) at the molecular level by employing antimicrobial resistance profiling, virulence genes profiling, and microbial typing approach using ERIC PCR. Based on multiple antibiotic resistance, overall, 19 antibiotic resistance patterns (R1-R19) were observed. Most of the strains (49/60) were detected to have the combinations of stx, eaeA, and hlyA genes and considered STEC/EPEC/EHEC. A total of 18 unique genetic profiles were identified based on ERIC-PCR fingerprints and most of the strains (13) belong to P1 whereas the least number of strains were showing profiles P7 and P8-P11 (one member each profile). The calculated values for Shannon index (H) for human, animal, and environment are 1.70, 1.82, and 1.78, respectively revealing the highest genetic diversity among the E. coli strains of animal origin. The study revealed that drug-resistant pathogenic E. coli strains could be transmitted bidirectionally among the environment, humans, and animals.

Assessment of serum biochemical derangements and associated risk factors of chronic kidney disease.

Objective: Chronic kidney disease and/or disturbance in renal excretory function may lead to nitrogenous waste collection beyond the term as well as derangements of several serum biochemicals. There is no previous study from Pakistan that reveals serum electrolyte derangements in confirmed chronic kidney disease (CKD) patients and other biochemicals associated with CKD. This study aims to examine the derangements of serum biochemicals and the association of several risk factors with CKD. Methods: The study enrolled 612 confirmed CKD patients with a glomerular filtration rate (GFR) < 15 ml/min that were treated as a part of the integrated care programme at Mayo Hospital Lahore (one of the largest hospitals in Pakistan). Serum biochemicals were estimated on AU 680 (Beckman Coulter) using the spectrophotometric technique. Results: All the CKD patients had elevated creatinine and urea levels, but only 63.4% were suffering from hyperuricemia. The incidence of diabetes and malnutrition assessed by serum albumin (hypoalbuminemia) was 27.4% and 72%, respectively. Among electrolyte disorders, hyperphosphatemia (71.8%) and hypocalcaemia (61.9%) were found to be more prevalent. Furthermore, gender, malnutrition, diabetes, hyperuricemia, and phosphorus and magnesium derangements were found to be statistically significant risk factors for CKD, whereas malnutrition and magnesium derangement were associated with hyperuricemia. Conclusion: It is imperative to improve dietary protein and monitor serum electrolyte concentration in renal dysfunction patients to slow the progression of CKD to end-stage renal disease (ESRD) and other serious complications.

Isolation and characterization of a highly effective bacterium Bacillus cereus b-525k for hexavalent chromium detoxification.

The chromate resistant Gram-positive Bacillus cereus strain b-525k was isolated from tannery effluents, demonstrating optimal propagation at 37 °C and pH 8. The minimum inhibitory concentration (MIC) test showed that B. cereus b-525k can tolerate up to 32 mM Cr6+, and also exhibit the ability to resist other toxic metal ions including Pb2+ (23 mM), As3+ (21 mM), Zn2+ (17 mM), Cd2+ (5 mM), Cu2+ (2 mM), and Ni2+ (3 mM) with the resistance order as Cr 6+ > Pb2+ > As3+ >Zn2+ >Cd2+ >Ni2+ >Cu2+. B. cereus b-525k showed maximum biosorption efficiency (q) of 51 mM Cr6+/g after 6 days. Chromate stress elicited pronounced production of antioxidant enzymes such as catalase (CAT) 191%, glutathione transferase (GST) 192%, superoxide dismutase (SOD) 161%, peroxidase (POX) 199%, and ascorbate peroxidase (APOX) (154%). Within B. cereus b-525k, the influence of Cr6+ stress (2 mM) did stimulate rise in levels of GSH (907%) and non-protein thiols (541%) was measured as compared to the control (without any Cr6+ stress) which markedly nullifies Cr6+ generated oxidative stress. The pilot scale experiments utilizing original tannery effluent showed that B. cereus b-525k could remove 99% Cr6+ in 6 days, thus, it could be a potential candidate to reclaim the chromate contaminated sites.

Successive use of microorganisms to remove chromium from wastewater.

Heavy metal pollution is a direct consequence of the extensive utilization of heavy metals in various industrial processes. The persistence and nondegradability of heavy metals cause them to bioaccumulate in nature, and when they come in direct contact with the pristine environment, they not only contaminate it severely but also pose dire consequences to the health of all living forms on earth, including humans. Chromium (Cr) is one of the heavy metals which has been extensively used in various industrial processes such as mining, alloy manufacturing, tanning of hides and skins, pigment production, etc. However, it is regarded as a priority pollutant due to its highly toxic, teratogenic, mutagenic, and carcinogenic nature, and the U.S. Environmental Protection Agency (EPA) also categorized it into group "A" human carcinogen. In contrast to water-soluble hexavalent chromium (Cr6+), its reduced form, trivalent chromium (Cr3+), is relatively benign and readily precipitated at environmental pH. Thus, bioremediation of Cr6+ through microorganisms including bacteria, yeast, and algae provides a promising approach to decontaminate a metal-polluted environment. This review describes an overview of the microbial reduction of Cr6+, resistance mechanism, and the antioxidant profiling exhibited by these microorganisms when exposed to Cr6+. It also describes the pilot-scale study of the successive use of bacterial, fungal, and algal strains and the subsequent use of microbially purified water for the cultivation of plant growth. Multiple metal-resistant microorganisms are a good bioresource for green chemistry to eradicate environmental Cr6+. KEY POINTS: • Hexavalent chromium (Cr6+) is highly toxic for living organisms including humans. • Microbial Cr resistance is mediated at the genetic, proteomic, and molecular levels. • Successive use of microorganisms is the best strategy to exterminate Cr6+from the environment.

Comparative behavior of two gram positive Cr6+ resistant bacterial strains Bacillus aerius S1 and Brevibacterium iodinum S2 under hexavalent chromium stress.

Bacillus aerius S1 and Brevibacterium iodinum S2 showed maximum growth at 37 °C and pH 8. B. aerius and B. iodinum could resist Cr6+ upto 30 and 35 mM and biosorption proficiency (q) of B. aerius S1 was 19, 27, 52 and 34 mM/g while for B. iodinum S2, it was 39, 50, 23 and 16 mM/g mM/g after 2, 4, 6 and 8 days of incubation. A pronounced rise in antioxidant enzymes activities was determined in B. aerius S1 i.e. POX (963%), CAT (717%), APOX (699%), SOD (683%), and GST (792%). However, in B. iodinum S2, relatively a minor increase was estimated. A significant GSH increase was determined in B. aerius S1 (364%) and B. iodinum S2 (663%) cultures under 2 mM Cr6+ stress. Pilot scale study demonstrated that both strains could reduce Cr6+ into Cr3+ within 6 days from the original tannery effluent with efficiency of 99%.

Bioconversion of hemicellulosic materials into ethanol by yeast, Pichia kudriavzevii 2-KLP1, isolated from industrial waste / Bioconversión de materiales hemicelulósicos en etanol por la levadura Pichia kudriavzevii 2-KLP1 aislada de residuos industriales

In the present work, a yeast strain Pichia kudriavzevii was identified on the basis of 18S rDNA, showing maximum growth at 30°C and pH 7.0. Among all the complex polysaccharides used, wheat bran proved to be the best substrate as indicated by the maximum growth of the yeast strain. The yeast isolate was capable of producing xylanase both intra-and extra-cellularly, the dominant form being extracellular. The maximum enzyme activity was determined at pH 5.0 and at 50°C. Na+, Mg2+ and Fe2+ presence caused a substantial increase in enzyme activity while a slight decrease (4.5%) was observed in the presence of Mn2+, Zn2+ and Cu2+. Pyruvate decarboxylase (PDC) and alcohol dehydrogenase (ADH) activities were assayed to confirm the presence of the ethanol pathway and PDC activity was much more pronounced (73%) compared to ADH activity (51%). The yeast strain can be employed to utilize hemicellulose containing agroindustrial residues for ethanol production.

En el presente estudio se identificó en aguas residuales de una zona industrial de Pakistán una cepa de la levadura Pichia kudriavzevii sobre la base del 18S ADNr, dicha cepa mostró un crecimiento máximo a 30 °C y a pH 7. Entre todos los sustratos de crecimiento evaluados para esta cepa, que incluyeron residuos industriales y medios definidos, el salvado de trigo demostró ser el mejor en función del crecimiento máximo alcanzado. Este aislado de levadura fue capaz de producir xilanasa intracelular y extracelular, esta última fue la forma predominante. Dicha capacidad enzimàtica mostró ser óptima a un pH de 5 y a 50°C. La presencia de Na+, Mg2+ y Fe2+ causó un incremento sustancial de la actividad enzimática, y hubo un ligero descenso (4,5%) en presencia de Mn2+, Zn2+ y Cu2+. Se evaluaron también las actividades de piruvato descarboxilasa y alcohol deshidrogenasa para confirmar la presencia de la vía del etanol. La actividad de la piruvato descarboxilasa fue mucho más pronunciada (73%) en comparación con la de alcohol deshidrogenasa (51%). Esta cepa de levadura puede emplearse para aprovechar los materiales hemicelulósicos de los residuos agroindustriales en la producción de etanol.

Bioconversion of hemicellulosic materials into ethanol by yeast, Pichia kudriavzevii 2-KLP1, isolated from industrial waste.

In the present work, a yeast strain Pichia kudriavzevii was identified on the basis of 18S rDNA, showing maximum growth at 30°C and pH 7.0. Among all the complex polysaccharides used, wheat bran proved to be the best substrate as indicated by the maximum growth of the yeast strain. The yeast isolate was capable of producing xylanase both intra- and extra-cellularly, the dominant form being extracellular. The maximum enzyme activity was determined at pH 5.0 and at 50°C. Na+, Mg2+ and Fe2+ presence caused a substantial increase in enzyme activity while a slight decrease (4.5%) was observed in the presence of Mn2+, Zn2+ and Cu2+. Pyruvate decarboxylase (PDC) and alcohol dehydrogenase (ADH) activities were assayed to confirm the presence of the ethanol pathway and PDC activity was much more pronounced (73%) compared to ADH activity (51%). The yeast strain can be employed to utilize hemicellulose containing agroindustrial residues for ethanol production.

Oxidative Stress, Chromium-Resistance and Uptake by Fungi: Isolated from Industrial Wastewater

ABSTRACT Trichosporon asahii and Rhodotorula mucilaginosa isolated from wastewater effluents were identified as chromium-resistant yeasts. Cr(VI) concentrations at 8 mM and 6 mM were inhibitory for R. mucilaginosa and T. asahii. Remarkably elevated GSH (69.88 ± 10.01) and GSSG (11.24 ± 0.96) was observed under metal stress in T. asahii as compared to R. mucilaginosa GSH (18.95 ± 3.19) and GSSG (3.7 ± 2.74) mM g-1 8 level. Statistical analysis revealed significantly higher GSH/GSSG ratio in both strains. NPSH (29.84 ± 0.54) level in T. asahii was much higher than in R. mucilaginosa (6.05 ± 0.24). Chromate reductase (ChR) was assayed and its activity was optimum at 50°C (pH 6) in T. asahii while R. mucilaginosa showed higher activity at 30°C (pH 7). Activity of both ChRs was enhanced in the presence of Mg, Na, Co and Ca but strongly inhibited by Hg cations. Cr(VI) uptake capabilities were ranged between 43-97% in R. mucilaginosa and 35-88% in T. asahii. One dimensional electrophoresis revealed enriched bands of cysteine rich metallothioneins suggesting some differential proteins could be overexpressed under Cr(VI) stress.