Standard Laboratory Protocols Research Articles

Physical and Chemical Properties of Soil of Different Villages of District Solan, Himachal Pradesh, India

The research was conducted to assess the physical and chemical properties of soils from twenty-one villages of Solan district, Himachal Pradesh within its geographical coordinates ranging from the latitude 30°44’53” to 31°22’01” N and longitude 76°36’10” to 77°15’14” E, during 2023-2024, India. The villages represent diverse landforms, including contour farming, plains, and slopes. Soil samples were collected from two depths, 0-15 and 15-30 cm, and analysed them using standard laboratory protocols. The predominant soil texture was sandy loam, bulk density ranges from 1.22 to 1.39 Mg m-3, particle density from 2.42 to 2.67 Mg m-3, and percent pore space varied between 40.50% and 46.68%, while the percent water holding capacity ranges from 37.65% to 45.53%. Soil pH ranged from 6.61 to 7.47. Electrical conductivity was measured between 0.18 and 0.34 dS m-1, indicating non-saline conditions. Organic carbon content was medium 0.30% to high 0.61%. The available nitrogen content ranged from 232.68 to 279.64 Kg ha-1, available phosphorus from 37.23 to 64.54 Kg ha-1, and available potassium from 262.14 to 365.34 Kg ha-1. These findings provide valuable insights into the soil health of the region, with implications for agricultural practices and land management.

The effect of soil moisture content and soil texture on fast in situ pH measurements with two types of robust ion-selective electrodes

Abstract. In situ soil pH measurements with ion-selective electrodes (ISEs) are receiving increasing attention in soil mapping for precision agriculture as they can avoid time-consuming sampling and off-site laboratory work. However, unlike the standard laboratory protocol, in situ pH measurements are carried out at lower and varying soil moisture contents (SMCs), which can have a pronounced effect on the sensor readings. In addition, as the contact with the soil during in situ measurements should be relatively short, effects of soil texture could be expected because texture controls the migration of protons to the electrode interface. This may be exacerbated by the fact that the electrodes used for in situ measurements are made of less sensitive but more robust materials as compared to the standard glass electrode. Therefore, the aim of the present study was to investigate the effect of soil moisture and soil texture on pH measurements using robust antimony and epoxy-body ISEs pressed directly into the soil for 30 s. The SMC was gradually increased from dry conditions to field capacity. A wide range of soil texture classes were included, with sand, silt, and clay contents ranging from 16 % to 91 %, 5 % to 44 %, and 4 % to 65 %, respectively. An exponential model was fitted to the data to quantify the relationship between SMC and pH. The results show that an increase in SMC causes a maximum increase in pH of approximately 1.5 pH units, regardless of the type of pH ISE used. Furthermore, for sandy soil textures, a rather linear relationship between pH and SMC was observed, whereas, with decreasing mean particle diameter (MPD), the model had a pronounced exponential shape, i.e., a greater pH increase at low SMC and a plateau effect at high SMC. With increasing SMC, the pH values asymptotically approached the standard pH measured with a glass electrode in 0.01 M CaCl2 (soil : solution ratio of 1:2.5). Thus, at high SMC, subsequent calibration of the sensor pH values to the standard pH value is negligible, which may be relevant for using the sensor pH data for lime requirement estimates. The pH measurement error decreases exponentially with increasing soil moisture and increases with decreasing MPD. Using a knee point detection, reliable pH values were obtained for SMC > 11 %, irrespective of the pH ISE used. An analysis of the regression coefficients of the fitted exponential model showed that the maximum pH increase also depends on soil texture; i.e., the influence of soil moisture variation on the pH value increases with decreasing MPD. Moreover, the concavity of the exponential curve increases with decreasing MPD.

Seasonal variation and uptake pattern of heavy metals in maize

The detrimental impact of heavy metals on human health necessitates a focus on their contamination in the food chain. Heavy metals possess carcinogenic, mutagenic, teratogenic, and endocrine-disrupting properties. The objective of the study was to ascertain how Zn, Pb, Cd, Ni, Co, Cu, Cr, and Hg are accumulated in the roots and grains of maize samples cultivated under the Kano River Irrigation Project during both wet and dry seasons production and subsequently compare the levels of these heavy metals with the recommended permissible limits established by FAO/WHO. Additionally, the research sought to employ Translocation Factors to examine the uptake pattern of heavy metals in maize during the two growing seasons. Maize samples were procured from 10 different locations within the irrigation scheme during 2022 wet season and 2023 dry season harvests. The samples were prepared using standard laboratory protocols and heavy metals were determined using atomic absorption spectrophotometer. The heavy metals accumulation in maize grains was found in the order Zn>Cu>Pb>Ni>Co during wet season production. Cd, Cr, and Hg were not detected in samples collected during the wet season. It is only the concentration of Zn that exceeded the permissible limit in the samples collected during wet season production. The accumulation order during the dry season was Zn>Cu>Cr>Pb>Ni>Co>Cd. Hg was not detected in all the maize grain samples. The concentrations of Zn, Pb, Cd and Cr exceeded permissible limits in maize grains samples collected during dry season production.

A comparative study on the detection of Mycobacterium leprae DNA in urine samples of leprosy patients using Rlep-PCR with other conventional samples.

Mycobacterium leprae causes leprosy that is highly stigmatized and chronic infectious skin disease. Only some diagnostic tools are being used for the identification M. leprae in clinical samples, such as bacillary detection, and histopathological tests. These methods are invasive and often have low sensitivity. Currently, the PCR technique has been used as an effective tool fordetecting M. leprae DNA across different clinical samples. The current study aims to detect M. leprae DNA in urine samples of untreated and treated leprosy patients using the Rlep gene (129 bp) and compared the detection among Ridley-Jopling Classification. Clinical samples (Blood, Urine, and Slit Skin Smears (SSS)) were collected from leprosy and Non-leprosy patients. DNA extraction was performed using standard laboratory protocol and Conventional PCR was carried out for all samples using Rlep gene target and the amplicons of urine samples were sequenced by Sanger sequencing to confirm the Rlep gene target. The M. leprae DNA was successfully detected in all clinical samples across all types of leprosy among all the study groups using RLEP-PCR. Rlep gene target was able to detect the presence of M. leprae DNA in 79.17% of urine, 58.33% of blood, and 50% of SSS samples of untreated Smear-Negative leprosy patients. The statistical significant difference (p = 0.004) was observed between BI Negative (Slit Skin Smear test) and RLEP PCR positivity in urine samples of untreated leprosy group. The PCR positivity using Rlep gene target (129 bp) was highest in all clinical samples among the study groups, across all types of leprosy. Untreated tuberculoid and PNL leprosy patients showed the highest PCR positivity in urine samples, indicating its potential as a non-invasive diagnostic tool for leprosy and even for contact screening.

Microbiological Methods Used in the Enterics for Global Health Shigella Surveillance Study.

Shigella is a major cause of diarrhea in young children worldwide. Multiple vaccines targeting Shigella are in development, and phase 3 clinical trials are imminent to determine efficacy against shigellosis. The Enterics for Global Health (EFGH) Shigella surveillance study is designed to determine the incidence of medically attended shigellosis in 6- to 35-month-old children in 7 resource-limited settings. Here, we describe the microbiological methods used to isolate and identify Shigella. We developed a standardized laboratory protocol for isolation and identification of Shigella by culture. This protocol was implemented across all 7 sites, ensuring consistency and comparability of results. Secondary objectives of the study are to determine the antibiotic resistance profiles of Shigella, compare isolation of Shigella from rectal swabs versus whole stool, and compare isolation of Shigella following transport of rectal swabs in Cary-Blair versus a modified buffered glycerol saline transport medium. Data generated from EFGH using culture methods described herein can potentially be used for microbiological endpoints in future phase 3 clinical trials to evaluate vaccines against shigellosis and for other clinical and public health studies focused on these organisms.

Occurrence and Antimicrobial Resistance of Staphylococcus aureus Isolated from Healthy Pet Rabbits.

Background: Staphylococcus aureus is a ubiquitous microorganism and an opportunistic pathogen responsible for numerous diseases in humans and animals, characterized by different clinical pictures with acute or subacute course. S. aureus, due to its great adaptability and versatility in terms of infections and hosts, can be considered a relevant pathogen because of the harmful effects on animal health and its potential for transmission from animals to humans and vice versa. In recent years, a marked increase in multidrug-resistant S. aureus has been reported, posing a serious threat for disease management, food safety, and animal and human health as they limit available therapeutic options. In light of a growing interest of the scientific community for this micro- organism and considering the limited data availability on the prevalence of this pathogen in pet rabbits, the purpose of this research was to evaluate the presence of S. aureus in pet rabbits. Materials and Methods: From November 2021 to December 2022, nasal swabs were collected from 50 pet rabbits from private households in the Campania Region, southern Italy, and underwent analysis for S. aureus detection. Samples were enriched in broth, then inoculated onto nutrient and selective media, including Blood agar base supplemented with 7% sheep blood and Baird-Parker Agar Base, following standard laboratory protocols. Incubations in aerobic conditions at 37°C were performed for 24/48h for colony identification. Antimicrobial susceptibility testing for all S. aureus isolates was conducted using the disc diffusion method. Results: Our results reported the presence of S. aureus in 16/50 (32%) rabbits examined, showing high levels of phenotypic resistance to different antibiotics, in particular penicillin 10U (81.2%) and erythromycin 15 μg (62.5%). Conclusion: The study demonstrated that pet rabbits represent a significant reservoir of S. aureus and contributes to the knowledge on the phenotypic antimicrobial resistance of these bacteria in rabbits raised in a domestic environment.

Phenotypic characterization of ESBL-producing urinary isolates of E. coli and Klebsiella spp. in a tertiary care children's hospital in Nepal

BackgroundThe production of extended-spectrum beta-lactamases (ESBLs) among uropathogens, particularly E. coli and Klebsiella spp., poses a severe public health concern. This study explored the epidemiology of ESBL-producing E. coli and Klebsiella spp. isolated from urine samples obtained at a tertiary care children's hospital in Nepal.MethodsA cross-sectional study was conducted from August 2016 to February 2017. A total of 745 clean catch urine samples were obtained from pediatric patients under the age of 13 and subjected to semiquantitative culture. E. coli and Klebsiella spp. were identified using standard laboratory protocols. Antibiotic susceptibility testing was performed using the Kirby-Bauer disc diffusion method, and ESBL producers were phenotypically identified using the combined disk method.ResultsAmong the bacterial isolates, E. coli predominated, accounting for 139 (81.8%) positive cases. Notably, E. coli showed high susceptibility to nitrofurantoin, with 117 (84.2%) isolates being susceptible. Meanwhile, K. pneumoniae showed high susceptibility to gentamicin, with 21 (91.3%) isolates being susceptible. Of the 163 isolates of E. coli and Klebsiella spp., 62 (38.0%) were identified as multidrug-resistant (MDR), with 42 (25.8%) confirmed as phenotypic ESBL producers. Remarkably, all 41 (100%) ESBL-producing E. coli isolates were susceptible to imipenem.ConclusionsThe prevalence of ESBL producers among E. coli and K. pneumoniae isolates from pediatric patients underscores the importance of antimicrobial stewardship. Nitrofurantoin and gentamicin emerge as effective empirical treatment choices against these pathogens in children. However, the high rates of multidrug resistance and ESBL production highlight the necessity for routine surveillance, and early detection strategies to manage such infections effectively.

In Vitro Assessment of Antimicrobial Activity of Novel Fluoroquinolone, Levonadifloxacin (WCK 771) Against Multi-Drug-Resistant Clinical Isolates from Cancer Patients in India.

Introduction: Rapid increase in antimicrobial-resistance is leading to urgent need for newer broad-spectrum antimicrobials. Therefore, we have evaluated the antimicrobial résistance spectrum of India-discovered novel antibiotics (levonadifloxacin) against clinical isolates recovered from cancer patients. Materials and Methods: The study was conducted in the microbiology department, over a period of 1 year between May 2021 and June 2022 and 374 consecutive and nonduplicate Gram-positive (GPC) and MDR Gram Negative Bacteria (GNB) isolate were analyzed from 3,880 cancer patients in study. The identification and antimicrobial sensitivities of bacterial isolates were performed according to standard laboratory protocols by using automated identification system (VITEK-2-8.01; BioMérieux, Germany). The activity of levonadifloxacin and comparator antibiotics was evaluated using disk diffusion methods as per Clinical and Laboratory Standards Institute 2022 guidelines. Results: The mean age of the patients were 51.6 ± 14.59 years with male: female ratio of 1.2:1. The prevalence of GPC was 167 (44.65%) and MDR-GNB was 207 (55.34%). The most common GPC was Staphylococcus aureus; 97 (58.08%) followed by Enterococcus species 66 (39.52%). In GNB, Escherichia coli; 93 (44.92%) was the most common followed by Klebsiella pneumoniae; 45 (21.73%). Levonadifloxacin susceptibility was present in 98.7% methicillin-resistant S. aureus and 96% methicillin-susceptible S. aureus and 77.1% Enterococcus-species. Additionally, all the fluoroquinolones-resistant S. aureus isolates were susceptible to levonadifloxacin (WCK-771) except one isolate. Also, levonadifloxacin-(WCK-771) exhibits 100% susceptibility fluoroquinolone susceptible GNB, such as E. coli, K. pneumoniae, Pseudomonas species, and Acinetobacter species. Interestingly, all fluoroquinolones-resistant Salmonella species and Stenotrophomonas maltophilla exhibited 100% susceptibility to levonadifloxacin (WCK-771). Conclusion: Levonadifloxacin (WCK-771) possesses potent activity against all the MDR Gram-positive pathogens including the coverage of susceptible Enterobacterales and MDR S. maltophilla and Burkholderia cepacia suggesting its potential utility in the management of polymicrobial infections.

Antimicrobial susceptibility pattern and mupirocin resistance among methicillin-resistant Staphylococcus aureus isolated from skin and soft-tissue infection patients attending a tertiary care hospital in North India

Background: Antimicrobial susceptibility among bacterial isolates varies from center to center. Mupirocin is a topical antibacterial antibiotic used to treat wound infections caused by methicillin-resistant Staphylococcus aureus (MRSA). Aim and Objectives: This study was conducted to know the prevalence of high and low levels of mupirocin resistance along with antimicrobial susceptibility in MRSA isolates from skin and soft-tissue infection in patients admitted to a tertiary care hospital in North India. Materials and Methods: A total of 97 non-repetitive isolates of MRSA from various pus samples over a period of 1 year were included in this study. These strains were identified as per standard laboratory protocols given by the Clinical Laboratory Standards Institute. High- and low-level mupirocin resistance of the isolates was tested by using mupirocin discs of 200 μg and 5 μg, respectively. Mupirocin strips of 256 μg and 512 μg were used for the E-test to detect low-level and high-level mupirocin resistance, respectively. The isolates were reported as sensitive to mupirocin, with minimum inhibitory concentrations of <4 mg/L. Results: Overall, 9.27% of mupirocin resistance was found, of which 2.06% of isolates were high-level resistant to mupirocin, while 7.21% had low-level resistance. All MRSA strains were susceptible to vancomycin, followed by linezolid (94.84%), teicoplanin (91.75%), and fusidic acid (88.65%). Conclusion: Regular monitoring, good infection control practices, and proper awareness about utilizing mupirocin therapeutically and prophylactically may help prevent the emergence of mupirocin resistance in health-care facilities.

Combination-Lock SlipChip Integrating Nucleic Acid Sample Preparation and Isothermal LAMP Amplification for the Detection of SARS-CoV-2.

Nucleic acid analysis with an easy-to-use workflow, high specificity and sensitivity, independence of sophisticated instruments, and accessibility outside of the laboratory is highly desirable for the detection and monitoring of infectious diseases. Integration of laboratory-quality sample preparation on a hand-held system is critical for performance. A SlipChip device inspired by the combination lock can perform magnetic bead-based nucleic acid extraction with several clockwise and counterclockwise rotations. A palm-sized base station was developed to assist sample preparation and provide thermal control of isothermal nucleic acid amplification without plug-in power. The loop-mediated isothermal amplification reaction can be performed with a colorimetric method and directly analyzed by the naked eye or with a mobile phone app. This system achieves good bead recovery during the sample preparation workflow and has minimal residue carryover from the lysis and elution buffers. Its performance is comparable to that of the standard laboratory protocol with real-time qPCR amplification methods. The entire workflow is completed in less than 35 min and the device can achieve 500 copies/mL sensitivity. Thirty clinical nasal swab samples were collected and tested with a sensitivity of 95% and a specificity of 100% for SARS-CoV-2. This combination-lock SlipChip provides a promising fast, easy-to-use nucleic acid test with bead-based sample preparation that produces laboratory-quality results for point-of-care settings, especially in home use applications.

Comparative evaluation of slit-skin smear with punch biopsy for diagnosing leprosy in suspected patient attending a tertiary care hospital in peripheral West Bengal

Background: Leprosy usually remains underdiagnosed or over-diagnosed due to a high dependency on clinical expertise. Leprosy requires a simple easy-to-perform laboratory test that guides clinicians to come to a conclusion with clinical correlation. Punch biopsy is the gold standard but it requires a biopsy facility in a hospital setting. Molecular diagnosis is out of reach even in resource-rich setups due to the non-availability of commercial kits. In this context slit-skin smear (SSS) might be an easier alternative to guide clinicians for a quality diagnosis, especially in peripheral hospitals. Aims and Objectives: To determine diagnostic sensitivity and specificity of SSS with respect to punch biopsy. Materials and Methods: The present study is an observational cross-sectional study conducted in a teaching institute in peripheral West Bengal. Two samples from clinically suspected patients of leprosy were collected for SSS and punch biopsy. Tests were performed as per standard laboratory protocol. Positive cases were informed to the clinician and treated as per World Health Organization treatment guidelines. SSS was compared to a punch biopsy with respect to sensitivity, specificity, positive predictive value, and negative predictive value. Results: Twenty-five among 59 clinically suspected cases were positive by either two tests or only one test. Twenty-one were males and four were female. Thirty-six was the median age distribution in positive cases 19 cases were positive by both SSS and punch biopsy, five by punch biopsy only, and one by SSS only. Sensitivity and specificity were 79.17% and 97.14%, respectively. Conclusion: SSS is not inferior to punch biopsy even in paucibacillary cases. It is a simple easy-to-perform test but requires training and awareness in medical personnel to make it a front-line weapon to combat continuous leprosy transmission in the community.

Increasing colistin resistance among multidrug-resistant isolates in the recent era – An impending doom

Background: Colistin is an antibiotic with lipophilic and hydrophilic properties, effective against Enterobacterales and non-fermenter bacteria. Due to the emergence of multidrug-resistant Gram-negative bacilli, colistin is being once again reintroduced as a last resort for their treatment. Resistance is caused by mutations and some are plasmid-mediated. Aims and Objectives: The aim of the study was to determine antibiotic resistance patterns in Gram-negative bacilli and to detect colistin susceptibility patterns in multidrug-resistant Enterobacterales and Pseudomonas aeruginosa. Materials and Methods: This prospective cross-sectional study was carried out in the Department of Microbiology, Calcutta National Medical College and Hospital, using standard laboratory protocol and colistin susceptibility was determined by colistin broth disk elution (CBDE) method. Results: Out of a total of 80 patients infected with MDR isolates, 63.75% were male and 36.25% were female. The most common age group affected was 41–60 years. Multidrugresistant (MDR) bacteria were most commonly isolated from urine samples, and Klebsiella pneumoniae was the most common MDR bacteria isolated. Overall, colistin resistance was 7.5% and it was found most commonly in K. pneumoniae. The use of broad-spectrum antibiotics was the most common risk factor for the emergence of MDR bacterial infections, including colistin resistance. Conclusion: The present study detects colistin resistance using a simple method – “CBDE test” which can be done in resource-poor settings and can guide clinicians to rationally use the medication as it may be used as an alternative treatment option for “Super bugs” and can even be used as the “last resort” in certain scenario.

Characterization of tetA gene of Escherichia coli isolated from colibacillosis affected calves in Rangpur, Bangladesh

Anti-microbial resistance is a matter of threat in global public health nowadays as a great number of antibiotics have lost their potential. In this study, we tried to characterize the tetA gene that was involved in tetracycline resistance of Escherichia coli (E. coli) in colibacillosis affected calves. To perform the study, a total of 69 fecal samples from colibacillosis suspected diarrheic calves were collected randomly from different areas of Rangpur division in Bangladesh. Standard laboratory protocols were followed for isolation and identification of E. coli. Then bacterial sensitivity was tested by culturing the bacteria in Mueller Hinton agar by following the protocol of Kirby-bauer disc diffusion method. Tetracycline resistant gene was specified by using tetA primer by searching tetA gene and the resistant gene was sequenced by sanger sequencing method. A phylogenetic tree of nucleotide sequences of tetA gene of E. coli was formulated using NCBI blast to characterize the gene. From 69 fecal samples, 54 were positive for E. coli. In antibiogram study, 100% isolates of E. coli were resistant to amoxicillin, ampicillin, cefoxitin, erythromycin and imipenem and the nature against others are as follows: doxycycline 64.8%, co-trimethoprim 63%, tetracycline 59.3%, ceftriaxone 50%, gentamicin 16.7%, levofloxacin 14.8% respectively. This study noticed, about 56.25% samples (18 from 32) were resistant to tetracycline due to tetA gene and the gene was closely related to the corresponding gene of Klebsiella pneumoniae, Shigella sonnei, Proteus mirabilis and Citrobacter braakii strains besides E. coli. This study suggests determining other genes specific for tetracycline resistance in E. coli of calves in Rangpur for better understanding the nature of it.

In-Vitro antidiabetic potential of Piliostigma thonningii leaf column chromatographic fractions

Diabetes mellitus is a metabolic disease characterized by high blood sugar that profoundly affects human health and economy. Although there are a plethora of antidiabetic drugs in the market, the efficacy shown by some medicinal plants sometimes outperforms that of synthetic drugs while being more affordable and less toxic. This study was conducted to evaluate the anti-diabetic potential of column-chromatographic fractions of Piliostigma thonningii leaf. Phytochemical screening, column, and thin layer chromatography were respectively conducted following standard laboratory protocols. The in-vitro antidiabetic activity of the extracts was assessed using alpha-glucosidase and hemoglobin glycosylation inhibitory methods. A total of 175 column fractions (CF1-CF175) were eluted respectively. Thin layer chromatography protocols employed pooled a total of 11 (A-J) fractions. The result showed significant (P>0.05) inhibition of alpha-glucosidase in only fraction J2, which was significantly greater when compared to standard drug (Voglibose) and crude extract. Also, there was no significant hemoglobin glycosylation inhibitory effect in all fractions compared with crude extract and standard drugs (Voglibose). In conclusion, P. thonningii leaf exhibited alpha-glucosidase and hemoglobin glycosylation inhibitory potentials that further support its usage as a hypoglycemic agent.

Source, distribution, and risk assessment of polycyclic aromatic hydrocarbons in sediment and fish samples from River Owan, Edo State, Nigeria

Polycyclic aromatic hydrocarbons (PAHs) are persistent environmental contaminants that present several environmental risks including human health. The 16 priority PAHs including its 1-methylnaphthalene, and 2-methylnaphthalene were determined in sediment and fish samples (Clarias anguillaris and Oreochromis niloticus) of River Owan, Edo State, Nigeria using gas chromatography (GC) equipped with flame ionization detector (FID) and other standard laboratory protocols. The isomeric ratio was used for source diagnosis, sediment quality guidelines, and risk models of incremental lifetime cancer were used for risk assessment. 1-methylnaphthalene and 2-methylnaphthalene were most predominant in all sediment samples analysed. The ∑LMW PAHs ranged between 0.093—0.250 μg/kg; ∑HMW PAHs were 0.107—0.579 μg/kg. The sediment samples range for ∑PAHs was 0.280–0.810 μg/kg with concentration order of increase: SE5>SE4>SE3>SE6>SE1>SE2>SE7 for the seven sampling locations. The ∑PAHs for Oreochromis niloticus was 0.190 μg/kg, which is higher than the value of Clarias anguillaris 0.080 μg/kg, and these values were greatly lesser when compared to the European Commission limit of 12.00 μg/kg. The diagnostic ratio indicates that the sources are more pyrogenic than petrogenic, revealing combustion from grass, wood, and bush burning. Sediment quality assessment showed that the ∑PAHs were lower than the regulatory values of sediment quality guidelines (SQG) assessment suggesting no ecotoxicological effects on the benthic organisms in this area at present. The Incremental Life Cancer Risk results were in the range of 9.15 × 10−12—1.46 × 10−6 for children, and 7.78 × 10−12—1.76 × 10−6 for adults considering the three routes of exposure. The incremental life cancer risk assessment showed a negligible risk.

Ameliorating effect of Calotropis procera methanol leaf extract against oxidative stress and lipid peroxidation induced by fipronil in Wister albino rats

The exposure of humans to oxidative stress inducing chemicals is on the increase due to global industrialization. It has become vital to source for solution by searching for medicinal plants with antioxidant potential. This study was carried out to evaluate the toxicological effect and the antioxidant potential of Calotropis procera in fipronil induced Wister Albino rats. Qualitative and quantitative phytochemical and antioxidant screenings of the leaf extract were conducted using standard laboratory protocols. Acute oral toxicity was conducted using Lorke's method to determine the LD50 of the plant extract. The sub-chronic toxicity was evaluated using the Organisation for Economic and Cultural Development method. The in vivo antioxidant potential of the plant extracts was determined using standard methods. Qualitative phytochemical analysis revealed the presence of saponins, flavonoids, phenols, cardiac glycosides, steroids, terpenoids and coumarines. Quantitative analysis revealed the amounts of saponins (32.67%), cardiac glycosides (7.6 mg/g), phenols (28 mg GAE/g), flavonoids (7.37 mg/g) and steroids (21.53 μg/mg) that were found. The LD50 was calculated to be 2154 mg/kg. Sub-chronic toxicity studies revealed insignificant (p>0.05) increases in body weight. Insignificant (p>0.05) changes in haematological parameters were also observed across treated groups compared to normal control. Liver and kidney function parameters were significantly (p<0.05) altered at higher (dose) of extract compared to normal control. In vivo antioxidant activity of the extract revealed strong antioxidant potentials due to significant (p<0.05) alteration of enzymatic (SOD, CAT, GSH and MDA) and non-enzymatic (vit C and E) antioxidants in the extract treated groups when compared to fipronil treated and normal control groups. Histological screening revealed that the liver and kidney tissues were significantly (p<0.05) altered at higher (dose) of extract administration. In conclusion, the present study validates the antioxidant potential and hence ameliorating effect of C. procera methanol leaf extract from toxic effects of phenyl pyrazole insecticides.

Influence of sport type and gender on bone turnover markers in young athletes

BackgroundExercise is beneficial to bone health. However, little is known about the interaction effect of gender and sport type on bone turnover in young athletes. This study aimed to examine the influence of gender and sports categories (high, medium, and low impact) on bone turnover: reabsorption markers–osteocalcin, calcium, inorganic phosphate (IP), alkaline phosphatase (ALP), and resorption marker–cross-linked N-telopeptides of type 1 collagen (NTx) among a university’s undergraduate athletes.MethodsThe study was an ex-post facto design involving forty-seven purposively recruited gender- and sport-type-matched undergraduate athletes whose demographic characteristics and BMI were obtained. Participants’ 5 mL antecubital blood samples were collected and analysed for serum levels of osteocalcin, calcium, IP, ALP, and NTx using standard laboratory protocols, Bio-Tek spectrometer, and KC4 (3.3) software. Data were analysed using descriptive statistics and two-way ANOVA.ResultsThe study involved 24 females and 23 males (n = 47) aged 22.15 ± 3.35 years with an average BMI of 23.34 ± 4.66. There was no significant gender effect on the biomarkers. However, there was a significant effect of the sports category on IP (F = 4.307, p = 0.020), calcium (F = 6.807, p = 0.003), and ALP serum levels (F = 11.511, p < 0.001). Specifically, mid-impact sports participants had a higher IP than the low-impact group (mean difference [MD] = 0.81 mg/dL, p = 0.036). Low-impact had a higher calcium level than mid-impact (MD = 0.40 mg/dL, p = 0.022) and high-impact (MD = 0.49 mg/dL, p = 0.003). Conversely, low-impact had lower ALP than mid-impact (MD = − 11.13 U/L, p = 0.013) and high-impact (MD = − 17.44 IU/L, p < 0.001).ConclusionModerate to high-impact sports positively affected bone turnover in young athletes. However, gender had no significant impact.

Frequent, infinitesimal bottlenecks maximize the rate of microbial adaptation.

Serial passaging is a fundamental technique in experimental evolution. The choice of bottleneck severity and frequency poses a dilemma: longer growth periods allow beneficial mutants to arise and grow over more generations, but simultaneously necessitate more severe bottlenecks with a higher risk of those same mutations being lost. Short growth periods require less severe bottlenecks, but come at the cost of less time between transfers for beneficial mutations to establish. The standard laboratory protocol of 24-h growth cycles with severe bottlenecking has logistical advantages for the experimenter but limited theoretical justification. Here we demonstrate that contrary to standard practice, the rate of adaptive evolution is maximized when bottlenecks are frequent and small, indeed infinitesimally so in the limit of continuous culture. This result derives from revising key assumptions underpinning previous theoretical work, notably changing the metric of optimization from adaptation per serial transfer to per experiment runtime. We also show that adding resource constraints and clonal interference to the model leaves the qualitative results unchanged. Implementing these findings will require liquid-handling robots to perform frequent bottlenecks, or chemostats for continuous culture. Further innovation in and adoption of these technologies has the potential to accelerate the rate of discovery in experimental evolution.

Evaluation of Fatty Acid and the Composition of Six Different Species of Freshwater Fish in the North of Algeria

&#x0D; &#x0D; &#x0D; Background: Few studies have been published about the quality of freshwater fish in Algeria. This study determined the chemical composition and the fatty acid of six species of freshwater fish cultivated in the North region of Algeria (Nile tilapia, red tilapia, common carp, Algerian barb, crucian carp, and mirror carp) as well as the nutritional quality of the lipids in these freshwater fish species.&#x0D; Methods: One hundred and ten freshwater fish were randomly caught in the spring of 2021 from Achor Ali farm (Jijel), Beni-Haroun Dam (Mila), and EL-Agrem Dam (Jijel) from Algeria. Moisture, ash, protein, lipid, and fatty acids were measured according to standard laboratory procedures and protocols of previous studies. Statistical analysis was performed using ANOVA (XLSTAT 2014), and the pair wise comparison of the means was done by Tukeys test at the 5% significance level (p&lt;0.05).&#x0D; Results: Regarding freshwater fish species, fatty acid profiles were discovered to have 38.94 to 57.75% Saturated Fatty Acids (SFAs), 29.35 to 46.63% Monounsaturated Fatty Acids (MUFAs), and 6.79 to 26.55% Polyunsaturated Fatty Acids (PUFAs). Common carp was a rich resource of Eicosapentaenoic Acid and Docosahexaenoic Acid (EPA+DHA) (5.38%); the highest concentration of ω-3 was recorded in crusian carp (10.63%); and Nile tilapia contained significant levels of ω-6 PUFA. Results demonstrated that the examined freshwater fish species appeared to have a good nutritional value and be a source of important fatty acids with positive effects on consumer health. On the other hand, results revealed low levels of PUFAs.&#x0D; Conclusion: The examined freshwater fish species appeared to have a good nutritional value, but, it is important to provide diets rich in fatty acids, in particular PUFAs, to these freshwater fish to improve the nutritional quality of their lipids.&#x0D; &#x0D; &#x0D;

A retrospective investigation of Candida spp. and its antifungal susceptibility profile in cancer patients at a South Indian tertiary care hospital

Among the most frequently recognized pathogenic fungi, . from Clinical samples plays a vital role. In recent decades, poor antifungal therapy usage critically in immunocompromised patients has led to antifungal drug resistance among . Hence, expeditious and dependable species identification is crucial for the effective implantation of treatment strategies and the management of diseases. The purpose of the study was to look at the presence of in various Clinical samples taken from cancer patients who are being treated at a tertiary care hospital in South India. The primary objective of this investigation is to ascertain the susceptibility patterns shown by antifungal drugs. This retrospective investigation was undertaken between January 2022 to April 2023. The isolates were subjected to antifungal susceptibility (AFST) by both Conventional (disc diffusion assay) and Automated methods (VITEK-2 compact - AST YSO1). The susceptibility pattern was recorded against antifungal agents like Fluconazole (FLU), Voriconazole (VOR), Amphotericin B (AMB), and Caspofungin. . was identified by conventional method (Chrome ID) as per standard laboratory protocols and by an automated method using MALDI – TOF. Among 215 samples received from cancer patients suggestive of Candidiasis, albicans (64.6%) was the predominant isolate, and the lowest one was (2.25%). Male patients had a higher prevalence of (55.8%). Similarly, the prevalence was higher in patients over 60, with male patients having an incidence of 38.3% and female patients having 23%. By disk diffusion assay and showed 100% sensitivity to all antifungals used in this study. We conclude that the disc diffusion assay is a more cost-effective, user-friendly, and effective screening test than the VITEK 2, and it has the potential to be used as a workable method for Antifungal susceptibility test. The use of Chrome ID for identification of . is cost-effective and reliable. However, MALDI TOF has the advantage of rapidity, providing correct findings promptly. The timely delivery of diagnostic reports facilitates early diagnosis of patients. Therefore, it is recommended in this research to use MALDI TOF for the identification of . and the disc diffusion technique for antifungal susceptibility testing (AFST).