Staining Test Research Articles

An Magnetic-Targeting Nano-Diagnosis and Treatment Platform for TNBC.

In this experiment, we constructed a magnetic targeting nano-diagnosis and treatment platform of doxorubicin (DOX) combined with iron nanoparticles, and explored their application value and mechanism in the treatment of Triple Negative Breast Cancer (TNBC), as well as its new diagnosis and treatment mode in Magnetic Resonance Imaging (MRI). Hollow mesoporous nanoparticles (HFON) were synthesized by solvothermal method, and loaded the drug DOX (DOX@HFON) to treat TNBC. The experiments in vivo and in vitro were carried out according to the characteristics of the materials. In vitro experiments, the killing effect of the drug on cells was verified by cell viability CCK8, ROS generation level, LPO evaluation and flow cytometry; the MRI effect and targeted anti-tumor therapy effect were studied by in vivo experiments; then the tumor tissue sections were detected by Ki-67, CD31, ROS, LPO and TUNEL immunofluorescence detection; H&E staining and blood biochemical tests were used to evaluate the biosafety of the materials. Through a series of characterization tests, it is confirmed that the nano-materials prepared in this experiment have positive drug loading properties. MDA-MB-231 cells had great phagocytic ability to DOX@HFON under Confocal Laser Scanning Microscope (CLSM). Experiments in vitro confirmed that DOX and Fe were released and concentrated in cells, and a large number of ROS production and induction of LPO were detected by DCFH-DA and C11-BODIPY probes in cells. Apoptosis experiments further confirmed that DOX@HFON induced apoptosis, autophagy and ferroptosis. In the vivo experiment, the anti-tumor therapy effect of MAGNET@DOX@HFON group was the most significant, and in MRI also proved that the drug had great tendency and imaging ability in tumor tissue. The new magnetic targeting nano-diagnosis and treatment platform prepared in this experiment is expected to become a new treatment model for TNBC.

Bacteriological study of the cerebrospinal fluid(CSF) of children with meningitis in Baghdad

The study included 92 samples of CSF taken from children aged 1day to 10 years with meningitis. The study was conducted in the hospitals of Medical City in Baghdad. The samples were obtained by a specialized staff. Conventional direct Gram stain and culture tests were performed. The study sample were categorized into three group a (1day to 2 years), (2 to 6 years) and (6 to 10 years).
 The results of the culture test showed that 62 samples out of 92 samples were free of bacteria while 30 samples contained bacteria (43% ). Among the positive culture group , 43% were gram positive bacteria and (57%) were Gram negative bacteria.
 Escherichia coli (43.4 %) was the predominant organism. The second bacterial agent is Staphylococcus aureus (20%) followed by Streptococcus pneumonia (13, 3%). The fourth bacterial isolates were Streptococcus agalactia (10%) and Neisseria meningitidis (10 %).The fifth bacterial agent was Klebsiella pneumonia (3.3%).
 The most common age group for bacterial meningitis was the first age group (1 to 2 years old) with Escherichia coli being the main bacterium (40%).

SiRNA-E6 sensitizes HPV-16-related cervical cancer through Oxaliplatin: an in vitro study on anti-cancer combination therapy

BackgroundPersistent infection with high-risk Human papillomaviruses (HPV), such as hr-HPV-16 and hr-HPV-18, lead to cervical cancer, the fourth most common cancer in the world. In the present study, we investigated the alteration of E6 oncogene expression by E6-specific short interfering RNA (siRNA) combined with Oxaliplatin.MethodsThe cervical cancer cell line, CaSki, was transfected with E6-siRNA, then treated with Oxaliplatin. The cellular genes, such as p53, MMP9, Nanog, and caspases expression, were assessed by quantitative real-time PCR. The cell death rate, cell cycle, and cell viability were assessed by Annexin V/PI staining, DAPI staining, and MTT test, respectively. Furthermore, colony formation assay and scratch test determined the stemness ability and cell metastasis, respectively.ResultsCombination therapy increased the re-expression of genes involved in the p53-dependent apoptosis pathway (increase in apoptosis to 44.2%), and reduced stemness and metastasis ability compared to either siRNA or Oxaliplatin monotherapy. Together, our results demonstrate that E6-siRNA and Oxaliplatin combination increased the cervical cancer cells’ sensitivity to Oxaliplatin and decreased the survival rate, proliferation, and metastasis, and consequently escalated apoptosis rate, induced cell cycle arrest in the sub-G1 stage, and reduced the chemotherapy drug dosage.ConclusionInhibition of E6 oncogene expression and subsequent E6-siRNA with Oxaliplatin combination therapy could be a novel strategy for cervical cancer treatment.Graphical

Dry eye in primary care: the relationship between digital display device usage and dry eye syndrome (DES) in medical students.

As digital display devices become more and more indispensable, associated health problems have started to increase. We aimed to investigate the frequency and relationship of digital display device usage behaviours and dry eye syndrome (DES) in medical students. An observational, cross-sectional study was conducted on medical school students who voluntarily participated in our study online, about the duration and variety of their digital screen use, usage patterns, and protective measures, together with the Ocular Surface Disease Index (OSDI) questionnaire. Where necessary, the Schirmer-I, tear break-up time, and fluorescein staining tests were carried out. Of the 255 participants, 75.3% (n = 192) were female, 63.1% (n = 161) had no refractive vision defects, and 45.9% used smartphones for more than 5 h a day. According to the OSDI scores for DES screening, only 36.1% of the participants were normal, 17.3% had mild DES, 20.4% had moderate DES, and 26.3% had severe DES. A positive correlation was found between digital display usage time in hours per day and a high OSDI score. As the distance between the eye and the screen decreased, the DES score increased, which is significant for computer, e-book, and tablet use (P < 0.005). The DES rate of the 25 participants who underwent further evaluation was 40% for the tear break-up time test and 52% for the fluorescein staining test. The high prevalence of DES due to digital screen use among medical school students highlights the importance of DES screening as a preventive measure in family medicine.

Dermatofibrosarcoma protuberans challenges: a case series and review of the literature

BackgroundDermatofibrosarcoma protuberans (DFSP) is a rare variant of skin sarcoma which is characterized by proliferation of spindle cells in a storiform pattern. Although it is mostly benign in its primary stages, it can cause a high burden of morbidity unless it is thoroughly excised.Case presentationHere, we review six cases of DFSP which were characterized by skin lesions in various parts of the body. Patients were from 26 to 51 years old; four were Asian men and two were Asian women. Wide surgical excision was performed for all these patients and no extra treatment was considered. Samples were studied by hematoxylin and eosin (H&E) staining and immunohistochemical (IHC) tests. Only one of our patients experienced recurrence after the initial surgery.ConclusionDetermining the best surgical method is still a dilemma in the treatment of DFSP lesions. There are numerous studies to prove the efficacy of various surgical interventions. Although DFSP is not commonly known as a malignant skin lesion, delay in treatment will have a catastrophic impact on patients’ lives. Thus, applying an in-time surgical method (wide local excision in our cases) in treating DFSP is crucial in preventing recurrence as well as decreasing the morbidity burden of DFSP.

Diversity, antimicrobial production, and seasonal variation of honey bee microbiota isolated from the honey stomachs of the domestic honey bee, Apis mellifera

The antimicrobial nature of honey and its related apiological origins typically focus on basic chemical analysis without attempting to understand the diversity of the microbial component. The antibacterial activity, chemical characterization, and diversity of bacteria isolated from Apis mellifera honey stomachs and hive honey collected throughout the honey production season are presented. After screening &amp;gt;2,000 isolates, 50 isolates were selected and characterized by 16S rRNA gene homology, Gram stain, catalase and protease tests, as well as for antibacterial activity against select indicators. Antibacterial-producing isolates were predominantly from the Pseudomonas, Paenibacillus, Lonsdalea, Serratia, and Bacillus genera. Isolates collected from honey stomachs in April displayed the highest level of activity (27%). While April isolates did not demonstrate activity against the Gram-negative bacteria tested. Whereas 59% of July isolates, 33% of September isolates, and 100% of the honey isolates did. The predominant honey stomach isolates were Pseudomonas spp. (April), Paenibacillus polymyxa (July, Sept.), and Lonsdalea iberica (Sept.). Chemical characterizations of the antimicrobial compounds show most to be antibiotic in nature with the minority being potential bacteriocins. This study offers the first glimpse into the variability and diversity of the bacteria/host interactions found within the honey stomach of the domestic honey bee while revealing a novel source of potentially beneficial antimicrobial compounds.

CTSB promotes sepsis-induced acute kidney injury through activating mitochondrial apoptosis pathway.

Acute kidney injury is a common and severe complication of sepsis. Sepsis -induced acute kidney injury(S-AKI) is an independent risk factor for mortality among sepsis patients. However, the mechanisms of S-AKI are complex and poorly understand. Therefore, exploring the underlying mechanisms of S-AKI may lead to the development of therapeutic targets. A model of S-AKI was established in male C57BL/6 mice using cecal ligation and puncture (CLP). The data-independent acquisition (DIA)-mass spectrometry-based proteomics was used to explore the protein expression changes and analyze the key proteomics profile in control and CLP group. The methodology was also used to identify the key proteins and pathways. S-AKI in vitro was established by treating the HK-2 cells with lipopolysaccharide (LPS). Subsequently, the effect and mechanism of Cathepsin B (CTSB) in inducing apoptosis in HK-2 cells were observed and verified. The renal injury scores, serum creatinine, blood urea nitrogen, and kidney injury molecule 1 were higher in septic mice than in non-septic mice. The proteomic analysis identified a total of 449 differentially expressed proteins (DEPs). GO and KEGG analysis showed that DEPs were mostly enriched in lysosomal-related cell structures and pathways. CTSB and MAPK were identified as key proteins in S-AKI. Electron microscopy observed enlarged lysosomes, swelled and ruptured mitochondria, and cytoplasmic vacuolization in CLP group. TUNEL staining and CTSB activity test showed that the apoptosis and CTSB activity were higher in CLP group than in control group. In HK-2 cell injury model, the CTSB activity and mRNA expression were increased in LPS-treated cells. Acridine orange staining showed that LPS caused lysosomal membrane permeabilization (LMP). CA074 as an inhibitor of CTSB could effectively inhibit CTSB activity. CCK8 and Annexin V/PI staining results indicated that CA074 reversed LPS-induced apoptosis of HK-2 cells. The JC-1 and western blot results showed that LPS inhibited mitochondrial membrane potential and activated mitochondrial apoptosis pathway, which could be reversed by CA074. LMP and CTSB contribute to pathogenesis of S-AKI. LPS treatment induced HK-2 cell injury by activating mitochondrial apoptosis pathway. Inhibition of CTSB might be a new therapeutic strategy to alleviate sepsis-induced acute kidney injury.

Molecular Characterization of Community- and Hospital- Acquired Methicillin-Resistant Staphylococcus aureus Isolates during COVID-19 Pandemic.

Methicillin-resistant Staphylococcus aureus (MRSA) is a drug-resistant superbug that causes various types of community- and hospital-acquired infectious diseases. The current study was aimed to see the genetic characteristics and gene expression of MRSA isolates of nosocomial origin. A total of 221 MRSA isolates were identified from 2965 clinical samples. To identify the bacterial isolates, the clinical samples were inoculated on blood agar media plates first and incubated at 37 °C for 18-24 h. For further identification, the Gram staining and various biochemical tests were performed once the colonies appeared on the inoculated agar plates. The phenotypic identification of antibiotic susceptibility patterns was carried out using Kirby-Bauer disk diffusion method by following the Clinical and Laboratory Standards Institute (CLSI) 2019 guidelines. The biofilm-producing potentials of MRSA were checked quantitatively using a spectrophotometric assay. All strains were characterized genotypically by SCCmec and agr typing using the specific gene primers. Furthermore, a total of twelve adhesion genes were amplified in all MRSA isolates. MRSA was a frequently isolated pathogen (44% community acquired (CA)-MRSA and 56% hospital acquired (HA)-MRSA), respectively. Most of the MRSA isolates were weak biofilm producers (78%), followed by moderate (25%) and strong (7%) biofilm producers, respectively. Prominent adhesion genes were clfB (100%), icaAD (91%), fib (91%), sdrC (91%) followed by eno (89%), fnbA (77%), sdrE (67%), icaBC (65%), clfA (65%), fnbB (57%), sdrD (57%), and cna (48%), respectively. The results of the current study will help to understand and manage the spectrum of biofilm-producing MRSA-associated hospital-acquired infections and to provide potential molecular candidates for the identification of biofilm-producing MRSA.

KHOL: What Impact on the Ocular Surface?

KHOL is usually applied along the edges of the eyelid, causing cosmetic particles to spread over the openings of the Meibomian glands. Two hundred womenwhose age varies between 18 and 25 years old were included in this cross-sectional descriptive study. Half use KHOL regularly and the other half rarely or never use it. The severity of ocular surface symptoms was investigated by the OSDI score and ocular surface evaluation by Schirmer’s test, fluorescein eye stain test and with a non-invasive break-up time (NIBUT). A moderate to severe dry syndrome was found in patients who used KHOL. The use of Khol causes disturbances in tear homeostasis and causes dry eye symptoms.

Risk factors, clinical indicators, and pathological findings of abomasal ulcers in tropical dairy buffaloes.

The current study aimed at identifying the risk factors and initial diagnostic aids for abomasal ulcers. The risk factor analysis confirmed a significant association (P < 0.05) of abomasal ulcers with concentrate-rich diets (OR, 4.795; CI, 1.212-15.974) and concurrent disorders (OR, 2.978; CI, 0.987-8.980), while the buffaloes in early lactation (OR, 2.777; CI, 0.703-10.972) showed a higher tendency (P = 0.078) for the disorder. The depressed demeanour, dark or black manure (melena), anemia, tachycardia, decreased milk production, anorexia, tachypnea, absence of rumination, abdominal guarding, kyphosis, and tachypnea were the most frequent clinical signs. Subjecting the abomasal fluid for cultural isolation, gram staining, and stormy clot fermentation test identified the presence of clostridium perfringes, while screening through uniplex PCR detected cpa toxin. The buffaloes affected with type-3 and 4 abomasal ulcers exhibited a higher peritoneal fluid to serum ratio of total protein, albumin, and glucose with a low (P < 0.01) serum-ascites albumin gradient (SAAG) concentration compared to reference values of healthy buffaloes. The first two principal components of PCA explained 54.50% of the total variances with lymphocytes, creatine kinase, and rumen chloride levels as the top contributors to dimension I, and albumin, total protein, sodium, and methylene blue reduction time (MBRT) for rumen liquor as the major contributors to dimension II. The vector plot revealed lymphocytopenia, decreased hemoglobin, hypoalbuminemia, hypokalemia, decreased rumen pH, neutrophilia, eosinophilia, leucocytosis, greater MBRT, and higher rumen chloride, serum creatine kinase, and blood urea nitrogen as the major indicators for abomasal ulcers. Histopathological studies revealed infiltration of inflammatory cells in the mucosa along with multifocal areas of necrosis, degeneration, and eroded muscle structure. The study projected a few high-scored clinical signs and extremely variable clinical indicators as initial diagnostic aids of abomasal ulcers, which can be confirmed by ultrasonography and peritoneal fluid examination.

The role played by ailanthone in inhibiting bone metastasis of breast cancer by regulating tumor-bone microenvironment through the RANKL-dependent pathway.

Introduction: Bone metastasis of breast cancer (BC) is a process in which the disruption of the bone homeostatic microenvironment leads to an increase in osteoclast differentiation. Ailanthus altissima shows an inhibitory effect on osteoclast differentiation. Ailanthone (AIL) refers to a natural compound isolated from Ailanthus altissima, a Chinese herbal medicine, and has effective anti-tumor activity in numerous cell lines. Its impact on bone metastases for BC is yet unclear. Methods: We measured the effect of AIL on MDA-MB-231 cells by wound healing experiments, Transwell and colony formation experiment. Using the Tartrate-resistant Acid Phosphatase (TRAP) staining tests, filamentous (F-actin) staining and bone resorption test to detect the effect of AIL on the osteoclast cell differentiation of the Bone Marrow-derived Macrophages (BMMs), activated by the MDA-MB-231 cell Conditioned Medium (MDA-MB-231 CM) and the Receptor Activator of Nuclear factor-κB Ligand (RANKL),and to explore its possibility Mechanisms. In vivo experiments verified the effect of AIL on bone destruction in breast cancer bone metastasis model mice. Results: In vitro, AIL significantly decrease the proliferation, migration and infiltration abilities of MDA-MB-231 cells at a safe concentration, and also reduced the expression of genes and proteins involved in osteoclast formation in MDA-MB-231 cells. Osteoclast cell differentiation of the BMMs, activated by MDA-MB-231 CM and RANKL, were suppressed by AIL in the concentration-dependent manner. Additionally, it inhibits osteoclast-specific gene and protein expression. It was noted that AIL inhibited the expression of the osteoclast differentiation-related cytokines RANKL and interleukin-1β (IL-1β) that were secreted by the MDA-MB-231 cells after upregulating the Forkhead box protein 3 (FOXP3) expression. Furthermore, AIL also inhibits the expression of the Mitogen-Activated Protein Kinase (MAPK), Phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT), and Nuclear factor-κB Ligand (NF-κB) signaling pathways, which then suppresses the MDA-MB-231CM-induced development of Osteoclasts. Conclusion: Our study shows that AIL blocks osteoclast differentiation in the bone metastasis microenvironment by inhibiting cytokines secreted by BC cells, which may be a potential agent for the treatment of BC and its secondary bone metastasis.

Establishment of acute liver failure model in Tibetan miniature pig and verified by dual plasma molecular adsorption system.

Acute liver failure (ALF) is a severe liver disease with high morbidity and mortality rates. Animal models are important for research on ALF. This study aimed to establish a reproducible, Tibetan miniature pig model of D-galactosamine-induced ALF and verify it using a dual plasma molecular adsorption system (DPMAS). Tibet miniature pigs were randomly divided into four groups (A, B, C, D) after catheterization. D-galactosamine (D-gal) at 0.45, 0.40, 0.35, and 0.35 g/kg body weight, respectively, was injected through the catheter. Group D was treated with DPMAS 48 h after D-gal administration. Vital signs and blood index values were recorded every 12 h after D-gal administration. H&E, TUNEL, Ki67, and Masson staining tests were performed. After D-gal administration, Tibetan miniature pigs developed different degrees of debilitation, loss of appetite, and jaundice. Survival times of groups A, B, C, and D were 39.7 ± 5.9, 53.0 ± 12.5,61.3 ± 8.1, and 61 ± 7 h, respectively. Blood levels of ALT, AST, TBIL, ammonia, PT, and inflammation factors significantly increased compared with baseline levels in the different groups (Ps < 0.05). Pathological results revealed a clear liver cell necrosis positive correlation with D-gal dose. However, DPMAS did not increase the survival time in ALF, ammonia, or liver cell necrosis. We successfully established a reproducible Tibetan miniature pig model of d-galactosamine-induced ALF, and we believe that a dosage of 0.35 g/kg is optimal.

Enterococcus faecium from chicken feces improves chicken immune response and alleviates Salmonella infections: a pilot study.

Probiotics reduce the emergence of antibiotic resistance in the livestock industry. Chicken feces are reservoirs of beneficial microbiomes. The aim of this study was to isolate putative probiotics from the intestinal contents of broiler chickens. Five fecal samples were collected from two poultry farms in Al-Ahsa, Saudi Arabia. Of the 11 morphologically distinct isolates from chicken feces (CF) samples, five isolates displayed positive reactions to Gram staining, catalase, and oxidase tests, and reacted negatively to a hemolytic assay. The isolates CF1, CF2, CF3, CF8, and CF11 were selected for further analysis of probiotic characterization, gastric survival capacity, antibiotic susceptibility, and antimicrobial activity against poultry infected with Salmonella enterica. CF2 and CF11 showed the highest hydrophobic values (> 51% hydrophobic nature). CF1, CF2, and CF11 showed potent antimicrobial activities. The active isolate CF2 was identified as Enterococcus faecium by 16s rRNA sequencing and showed a genetic similarity of 99.1%. An in vivo study was conducted using a chicken model. Enterococcus faecium-fed chickens showed an improved body weight and a lower mortality rate (17-34%). Salmonella enterica colony-forming unit (CFU) invasion in the spleen and thymus was significantly reduced in the E. faecium-fed chickens. The fecal S. enterica load was reduced from CFU 6.8 to 3.9/g in oral-administered E. faecium-fed chickens. Enterococcus faecium-fed chickens showed increased levels (P < 0.01) of butyric acid and reduced levels (P < 0.01) of intestinal interleukin 1 beta, C-reactive protein, and interferon gamma levels compared to those in the S. enterica-infected chicken group. In addition, E. faecium showed significant binding to Caco-2 epithelial cells in vitro and inhibited S. enterica colonization, indicating co-aggregation of E. faecium in epithelial cells. These results revealed that an E. faecium formulation could prevent bacterial infection and improve the quality of broiler chickens.

PHENOTYPIC DETECTION OF Escherichia coli PRODUCING Extended Spectrum Beta Lactamases (ESBLs) IN THE REPRODUCTIVE TRACT BALI COW

&#x0D; Escherichia coli bacteria that are resistant to β-lactam antibiotics and may be able to produce Extended-spectrum β-lactamase (ESBLs) have been found in the reproductive tract of Bali cattle. Escherichia coli that produce ESBL will cause antimicrobial resistance (AMR) which has become a global health problem. The purpose of this study was to detect phenotypically the presence of Escherichia coli that produces ESBL from the reproductive tract of Bali cattle. This research was conducted in February 2022 in Lando Village, Terara District, East Lombok Regency using 8 female Bali cattle on smallholder farms. Reproductive fluid samples were collected using a plastic sheet gun which was inserted into the Brain Infusion Heart (BHI) media and incubated at the Public Health and Calibration Laboratory of West Nusa Tenggara Province. The samples were then cultured in EMBA media for the isolation of Escherichia coli. The isolated Escherichia coli were then identified by gram staining and biochemical tests based on Bergey's manual of determinative bacteriology. The identified Escherichia coli were screened using the disc diffusion method using the antibiotics Penicillin G10U, Ceftazidime 30 μg, and Cefotaxime 30 μg followed by a confirmation test using the double disk synergy test (DDST) method. The results of the screening test showed that only 12.5% of Escherichia coli (1 out of 8 samples) were resistant to Penicillin G, but they had not been detected to produce ESBL phenotypically.

DETECTION OF ANTIBIOTIC RESISTANT IN Escherichia coli FROM THE REPRODUCTIVE TRACT OF BALI CATTLE ON SMALLHOLDER FARM

Escherichia coli originating from animals, humans, and the environment can spread antibiotic-resistant genes and can encourage antimicrobial resistance (AMR) which is a global health problem. The purpose of this study was to detect the presence of Escherichia coli which is resistant to several antibiotics from the reproductive tract of Bali cattle on smallholder farms on the island of Lombok. This research is a cross-sectional study conducted from March to June 2021 using 8 female Bali cattle that experience reproductive disorders on community farms in Lando Village, East Lombok Regency to collect their reproductive tract fluids using an artificial insemination plastic sheet gun. Reproductive fluid samples are placed on BHIB (Brain Heart Infusion Broth) medium. Escherichia coli cultures were carried out on Eosin Methylene Blue Agar (EMBA) and identified by Gram staining and biochemical tests conducted at the Public Health and Calibration Laboratory, West Nusa Tenggara Province. Antibiotic sensitivity test on isolated Escherichia coli was carried out by disc diffusion method using 5 antibiotics including Penicillin G 10U, Oxytetracycline 30 µg, Gentamicin 10 µg, Tetracycline 30 µg, and Cefotaxime 30 µg. The results showed that 2 (two) 25% Escherichia coli bacteria were successfully cultured on EMBA media and isolated from 8 samples of the reproductive fluids of Bali cattle that were collected. The results of the Escherichia coli sensitivity test to antibiotics found that Escherichia coli samples were 100% resistant to Penicillin G, 100% resistant to Oxytetracycline, 100% resistant to Gentamicin, and 50% resistant to Tetracycline, and 100% resistant to Cefotaxime from 2 isolated Escherichia coli. This explains that Bali cattle in community farms have the potential to spread Escherichia coli which has an impact on the emergence of AMR.

Diversity of Beneficial Bacteria in Tamhini Ghat Region of Western Ghats, M.S., India

The present study investigates that the isolation of beneficial bacteria from a soil sample collected from the Tamhini Ghat region of the Western Ghats. In October 2022, 15 soil samples were collected from Tamhini Ghat. Five different beneficial bacteria were isolated from these collected soil samples and characterized morphologically. The beneficial bacteria like Azotobacter spp., Bacillus spp., Azospirillum spp., Pseudomonas spp., and Frateuria spp. Isolation of these bacteria was carried out on a specific medium and observed after three days of the incubation period at 28±2 0 C temperature. These isolated bacteria were purified on slants prepared by using a specific medium and stored at 4 0 C for further study and observed under a microscope by performing a Gram staining test. These beneficial bacteria can be used for biofertilizer formulation in future studies. All over India, farmers are using chemical fertilizers, which directly impact beneficial soil microbe that leads to decreased soil health. For improving soil health, it is the need of the hour to integrate the use of chemical fertilizers along with biofertilizers as per the recommended dose of fertilizers (RDF) given by the agricultural universities for the specific crops.

Nasopharyngeal Carriage Rate of Streptococcus pneumoniae, Related Risk Factors, and Antibiotic Susceptibility Among Inmates in Hawassa Central Prison Institute: Hawassa, Sidama National Region, Ethiopia.

The bacterium Streptococcus pneumoniae commonly causes severe bacterial illnesses in both children and adults. S. pneumoniae is the most fatal bacterial pathogen, causing 50% of the deaths from lower respiratory infections in people of all ages. This study was aimed to determine a nasopharyngeal carriage rate of S. pneumoniae, related risk factors, and antibiotic susceptibility among prisoners in Hawassa Central Prison Institute Hawassa, Ethiopia. From July to August 2021, a cross-sectional investigation was carried out in Hawassa prison. All randomly chosen adult volunteers were a part of the study. In this study, about 330 different people were included. Within 3 h of collection, the swab specimen was inoculated onto 5% sheep blood agar; the inoculated medium was then incubated for 18-24 h at 35-37 °C in a CO2-enriched environment. Following incubation, the S. pneumoniae colonies were suspected, but the usual colony shape, alpha-hemolysis, Gram staining, optochin susceptibility, and bile solubility test allowed for confirmation. Antimicrobial susceptibility testing was carried out in accordance with the Clinical and Laboratory Standards Institute's guidelines. Overall carriage rate of S. pneumoniae among adult individuals were 41 (12.7%). The carriage rate of S. pneumoniae was affected by age classes that 39-48 years (OR = 4.82, 95%CI = 1.49-15.56, p = .009), 49-58 years (OR = 5.27, 95%CI = 1.27-21.89, p = .022), and greater than 58 years (OR = 4.36, 95%CI = 1.13-16.90, p = .033); cigarette smoking (OR = 3.41, 95%CI = 1.16-10.01, p = .026); and sharing beds (OR = 3.91, 95%CI = 1.27-12.07, p = .018). The majority of the isolates are susceptible for clindamycin (87.8%) and resistant for chloramphenicol (56%). Multidrug resistance was observed in 36.7%. The overall carriage rate of S. pneumoniae was 12.7%. Age, cigarette smoking, and sharing beds in the same prison room had a substantial impact on this nasopharyngeal carriage rate. Chloramphenicol (56%), erythromycin (41.5%), tetracycline (39%), and co-trimoxazole (34.1%) resistance were seen in the majority of S. pneumoniae isolates. Early detection, hygiene maintenance, and appropriate treatment are necessary.

Uropathogens and their antimicrobial-resistant pattern among suspected urinary tract infections patients in eastern Nepal: A hospital inpatients-based study.

Urinary tract infections are the primary factors that cause mortality and morbidity in patients with underlying comorbid conditions and are responsible for most hospital admissions worldwide. The study aims to identify the common bacterial uropathogens and determine their antimicrobial susceptibility pattern, including multidrug-resistant/extensively drug-resistant bacteria. The descriptive cross-sectional study was conducted among inpatients provisionally suspected of urinary tract infections in the medical ward of Koshi Hospital, Biratnagar, Nepal. Samples were inoculated in a cystine lysine electrolyte-deficient medium, and pure growth of significant bacteria was further subjected Gram staining, biochemical identification, and antimicrobial susceptibility testing as per laboratory standard procedure and Clinical Laboratory Standards Institute guidelines, respectively. Descriptive and inferential statistical analysis was performed to analyze the outcomes and a p-value < 0.05 was considered statistically significant. A total of 305 patients urine specimens were examined, of which 251 (82.29%) samples resulted in significant bacterial growth in the culture. Escherichia coli (62.94%) was the most predominantly isolated organism, followed by Klebsiella pneumoniae (12.35%), Staphylococcus aureus (9.16%), and Pseudomonas aeruginosa (8.76%). Among antimicrobials, colistin had shown absolute susceptibility (100%) toward gram-negative uropathogens followed by carbapenem and aminoglycosides in a majority of uropathogens. Escherichia coli was found to be the leading drug-resistant bacteria (70%) among uropathogens. The presence of multidrug-resistant/extensively drug-resistant bacteria uropathogens was found to be significantly associated with diabetes mellitus and those with combined antimicrobial therapies. Diabetic patients were twice (OR~2) more likely to colonize and develop uropathogens as compared to non-diabetics. Escherichia coli was the most common uropathogens followed by Klebsiella pneumoniae in urinary tract infection patients. The polymyxin group (colistin) of antimicrobials was found to be effective in all multidrug-resistant and extensively drug-resistant uropathogens. The study recommends the need of optimized antimicrobial stewardship program to develop effective strategies in the management of urinary tract infections in diverse healthcare settings.

&amp;lt;i&amp;gt;Parkia biglobosa&amp;lt;/i&amp;gt; Fruit Husks: Phytochemistry, Antibacterial, and Free Radical Scavenging Activities

Known for their role in the manufacture of eco-materials in buildings, Parkia biglobosa fruit husks are also used in folk medicine. The present study focuses on the metabolites content and antiradical, antibacterial activities of the hydroethanolic extract of P. biglobosa husks. Secondary metabolites were identified using staining and/or precipitation tests. The mineral content is determined according to the standard NF EN 14082. The antioxidant activity performed by 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging capacity, and antibacterial activity strain against three Gram-negative and two Gram-positive strains by microdilution for Minimal Inhibitory Concentration. Fruit husks contain tannins, anthocyanins, leuco-anthocyanins, anthraquinones, saponins, reducing compounds, sterols, terpenes, and 1.225% of potassium. The hydroethanolic extract of P. biglobosa fruit husks scavenges the DPPH radical with an EC50 = 64 μg/ml. The extract is more active in one Gram (Pseudomonas aeruginosa and Escherichia coli) with a minimum inhibitory concentration of 1.25 mg/mL than the three Gram + studied. This study showed that P. biglobosa fruit husks extract could be used for its antioxidant and antibacterial activities.

Molecular Detection and Multidrug Resistance of Shigella spp. Isolated from Wild Waterfowl and Migratory Birds in Bangladesh

Birds, especially wild waterfowl and migratory birds have the potential to carry antibiotic‐resistant bacteria, but their role in the dissemination of these resistant pathogens is still neglected in Bangladesh. To the best of our knowledge, this study was carried out for the first time in Bangladesh to isolate and determine the occurrence of multidrug‐resistant (MDR) Shigella spp. from fecal materials of wild waterfowl and migratory birds. A total of 80 fecal materials from wild waterfowl (n = 50) and migratory birds (n = 30) were screened to detect MDR Shigella isolates. Shigella spp. were isolated and identified by culturing, staining, and biochemical tests followed by polymerase chain reaction (PCR). A disk diffusion assay was employed to investigate antibiotic phenotypes, while the resistance genes were detected by PCR. Among the 80 samples, 15 (18.75%) were found positive for Shigella spp. by PCR, among which the occurrence rate of Shigella spp. was higher in migratory birds (20%, 6/30) than in wild waterfowl (18%, 9/50). By the disk diffusion test, 86.67% (13/15) of Shigella spp. isolates were found to be MDR in nature, including 93.33% of isolates resistant to imipenem. Moreover, frequent and moderate resistance was also observed against tetracycline (86.67%), azithromycin (80%), ampicillin (66.67%), ciprofloxacin and cotrimoxazole (40%), meropenem (26.67%), and streptomycin (13.33%). The bivariate analysis revealed a positive correlation between the resistance profiles of ciprofloxacin and cotrimoxazole, imipenem and tetracycline, tetracycline and ampicillin, and imipenem and azithromycin. Furthermore, the isolates had a multiple antibiotic resistance index of up to 0.47. Antibiotic resistance genes tetA and SHV were found in 69.23% and 50% of relevant antibiotic‐resistant Shigella spp. isolates, respectively. The present study suggests that wild waterfowl and migratory birds are reservoirs of MDR Shigella spp., which may have detrimental impacts on One Health components. We suggest keeping these birds under an AMR monitoring program to avoid the possibility of AMR contamination of the environment and its consequences in all health settings.