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AnteayerPLOS ONE Medicine&Health

Application of in-silico drug discovery techniques to discover a novel hit for target-specific inhibition of SARS-CoV-2 Mpro’s revealed allosteric binding with MAO-B receptor: A theoretical study to find a cure for post-covid neurological disorder

by Magdi E. A. Zaki, Sami A. AL-Hussain, Aamal A. Al-Mutairi, Abdul Samad, Vijay H. Masand, Rahul G. Ingle, Vivek Digamber Rathod, Nikita Maruti Gaikwad, Summya Rashid, Pravin N. Khatale, Pramod V. Burakale, Rahul D. Jawarkar

Several studies have revealed that SARS-CoV-2 damages brain function and produces significant neurological disability. The SARS-CoV-2 coronavirus, which causes COVID-19, may infect the heart, kidneys, and brain. Recent research suggests that monoamine oxidase B (MAO-B) may be involved in metabolomics variations in delirium-prone individuals and severe SARS-CoV-2 infection. In light of this situation, we have employed a variety of computational to develop suitable QSAR model using PyDescriptor and genetic algorithm-multilinear regression (GA-MLR) models (R2 = 0.800–793, Q2LOO = 0.734–0.727, and so on) on the data set of 106 molecules whose anti-SARS-CoV-2 activity was empirically determined. QSAR models generated follow OECD standards and are predictive. QSAR model descriptors were also observed in x-ray-resolved structures. After developing a QSAR model, we did a QSAR-based virtual screening on an in-house database of 200 compounds and found a potential hit molecule. The new hit’s docking score (-8.208 kcal/mol) and PIC50 (7.85 M) demonstrated a significant affinity for SARS-CoV-2’s main protease. Based on post-covid neurodegenerative episodes in Alzheimer’s and Parkinson’s-like disorders and MAO-B’s role in neurodegeneration, the initially disclosed hit for the SARS-CoV-2 main protease was repurposed against the MAO-B receptor using receptor-based molecular docking, which yielded a docking score of -12.0 kcal/mol. This shows that the compound that inhibits SARS-CoV-2’s primary protease may bind allosterically to the MAO-B receptor. We then did molecular dynamic simulations and MMGBSA tests to confirm molecular docking analyses and quantify binding free energy. The drug-receptor complex was stable during the 150-ns MD simulation. The first computational effort to show in-silico inhibition of SARS-CoV-2 Mpro and allosteric interaction of novel inhibitors with MAO-B in post-covid neurodegenerative symptoms and other disorders. The current study seeks a novel compound that inhibits SAR’s COV-2 Mpro and perhaps binds MAO-B allosterically. Thus, this study will enable scientists design a new SARS-CoV-2 Mpro that inhibits the MAO-B receptor to treat post-covid neurological illness.

Implementing HLA-B*58:01 testing prior to allopurinol initiation in Malaysian primary care setting: A qualitative study from doctors’ and patients’ perspective

by Wei Leik Ng, Norita Hussein, Chirk Jenn Ng, Nadeem Qureshi, Yew Kong Lee, Zhenli Kwan, Boon Pin Kee, Sue-Mian Then, Tun Firzara Abdul Malik, Fatimah Zahrah Mohd Zaidan, Siti Umi Fairuz Azmi

Introduction

Allopurinol, the first-line treatment for chronic gout, is a common causative drug for severe cutaneous adverse reactions (SCAR). HLA-B*58:01 allele was strongly associated with allopurinol-induced SCAR in Asian countries such as Taiwan, Japan, Thailand and Malaysia. HLA-B*58:01 screening before allopurinol initiation is conditionally recommended in the Southeast-Asian population, but the uptake of this screening is slow in primary care settings, including Malaysia. This study aimed to explore the views and experiences of primary care doctors and patients with gout on implementing HLA-B*58:01 testing in Malaysia as part of a more extensive study exploring the feasibility of implementing it routinely.

Methods

This qualitative study used in-depth interviews and focus group discussions to obtain information from patients with gout under follow-up in primary care and doctors who cared for them. Patients and doctors shared their gout management experiences and views on implementing HLA-B*58:01 screening in primary care. Data were coded and analysed using thematic analysis.

Results

18 patients and 18 doctors from three different healthcare settings (university hospital, public health clinics, private general practitioner clinics) participated. The acceptability to HLA-B*58:01 screening was good among the doctors and patients. We discovered inadequate disclosure of severe side effects of allopurinol by doctors due to concerns about medication refusal by patients, which could potentially be improved by introducing HLA-B*58:01 testing. Barriers to implementation included out-of-pocket costs for patients, the cost-effectiveness of this implementation, lack of established alternative treatment pathway besides allopurinol, counselling burden and concern about genetic data security. Our participants preferred targeted screening for high-risk populations instead of universal screening.

Conclusion

Implementing HLA-B*58:01 testing in primary care is potentially feasible if a cost-effective, targeted screening policy on high-risk groups can be developed. A clear treatment pathway for patients who test positive should be made available.

Cross-sectional study of attitudes toward online continuing dental education in Libya during the COVID-19 pandemic

by Ranya F. Elemam, Jamal M. El Swiah, Abduallah O. Durda, Nagwa N. Hegazy

During the COVID-19 pandemic, many educational institutions switched to e-learning educational platforms. This approach was essential but raised challenges, particularly in training practitioners for medical emergencies. This approach not only led to global challenges and a need for rapid adaptation, but also raised inequities across countries, with some facing far more technical challenges than others. In Libya, low investment in education technology and unpredictable internet connectivity limited its integration into schools and universities even before the pandemic. The current study reports feedback from an online continuing dental education (CDE) course for dental practitioners that was developed emergently during the pandemic and aimed to address the challenges posed by Libya’s internet environment. Participants were recruited through social media and received an 8-hour online CDE course consisting of three modules. Participants were invited to complete a pre-course demographic/informational survey on a Google form. After passing all modules, students were prompted to complete a post-course survey consisting of 23, five-point Likert scale questions. Respondents included 43 females (74.1%) and 15 males (25.9%). For ~50% of the cohort (n = 32), this was their first online clinical course. 87.9% of post-course participants rated the course as a positive learning experience, while 90.9% agreed their learning outcomes had been achieved. Most participants (97%) agreed the course instructor explained all concepts clearly. In total, 81.8% agreed that the technology effectively supported their learning. Most agreed that a clear demarcation between each course module existed and that the language and depth of the material were adequate. Some students reported technical difficulties, and 33.3% saw repetitions in the modules. However, all post-course respondents said they would recommend the online course to colleagues. Libyan dental practitioners showed high satisfaction levels towards the e-learning process, course content, instructors’ attitudes, and overall e-learning experience despite the inherent e-learning challenges posed in this country.

Understanding volunteerism among dental students and professionals to reach Saudi Arabia Vision 2030 goals

by Khalid Aboalshamat, Turki Alayyafi, Ghassan Elaiwa, Mishal Assayegh, Abdulmalk Alqaidi

Background

Volunteering can be defined as activities a person does for free to help another person or group. Saudi Vision 2030 has a target of one million volunteers from the country by 2030. The aim of this study was to find out the frequencies of the motives, barriers, and experiences of volunteering dental students and dentists in Saudi Arabia.

Materials and methods

In this cross-sectional study, 655 dental students and dentists from 37 cities around Saudi Arabia answered a questionnaire of 59 questions derived from previous studies. The questionnaire was distributed through social media. SPSS software was used to analyze the data, with p-value of 0.05 as significant. Chi-square was used for analytical statistics.

Results

The chance to learn in a health-related field (84.58%) was the most motivating factor to volunteer, and the least motivating factor was financial compensation (46.72%). Females had multiple significantly higher motives percentages than males (p Conclusions

A high percentage of dental students and dentists in Saudi Arabia engage in volunteer activities. Nevertheless, various impediments must be addressed to achieved the targeted key performance indicator of Saudi Arabia Vision 2030.

SRY gene isolation from teeth for forensic gender identification—An observational study

by Prathibha Prasad, Mohamed Jaber, Dinesh Y., Prathibha Ramani, Abdulrahman Arafat, Abdalla Khairy

Personal identification in forensics is possible with gender determination using DNA (deoxyribonucleic acid) analysis. DNA isolation from teeth samples subjected to extreme temperatures has been shown to predict the gender of the deceased. However, the literature lacks studies on DNA extracted from tooth samples exposed to freezing temperatures. This study aimed to isolate the SRY gene from the extirpated pulp of teeth that were subjected to varying temperatures for gender identification. Thirty teeth with vital pulps, divided into 3 groups were included in the study. Each group consisted of 5 male and 5 female tooth samples. The groups were exposed to diverse environmental factors for three weeks. Group 1: room temperature (R group); Group 2: high temperature (H group) and Group 3: freezing temperature (F group). Later, DNA was isolated from the pulp tissue, and the SRY gene was amplified using PCR (Polymerase Chain Reaction). The Sensitivity and Specificity of the results were analyzed. SRY gene detected in the study samples identified accurate gender with a 46.70% Sensitivity and 93.30% Specificity. Significant difference was found in the correlation between gene expression and gender among the three groups (p = 1.000). The study validates that dental pulp tissue can be a reliable source for DNA extraction. And SRY gene amplification from teeth exposed to diverse environmental conditions. Further investigations are required to validate its application in forensics.

Digital health interventions for cervical cancer care: A systematic review and future research opportunities

by Md Abdur Razzak, Muhammad Nazrul Islam, Md Shadman Aadeeb, Tasfia Tasnim

Background

Cervical cancer is a malignancy among women worldwide, which is responsible for innumerable deaths every year. The primary objective of this review study is to offer a comprehensive and synthesized overview of the existing literature concerning digital interventions in cervical cancer care. As such, we aim to uncover prevalent research gaps and highlight prospective avenues for future investigations.

Methods

This study adopted a Systematic Literature Review (SLR) methodology where a total of 26 articles were reviewed from an initial set of 1110 articles following an inclusion-exclusion criterion.

Results

The review highlights a deficiency in existing studies that address awareness dissemination, screening facilitation, and treatment provision for cervical cancer. The review also reveals future research opportunities like explore innovative approaches using emerging technologies to enhance awareness campaigns and treatment accessibility, consider diverse study contexts, develop sophisticated machine learning models for screening, incorporate additional features in machine learning research, investigate the impact of treatments across different stages of cervical cancer, and create more user-friendly applications for cervical cancer care.

Conclusions

The findings of this study can contribute to mitigating the adverse effects of cervical cancer and improving patient outcomes. It also highlights the untapped potential of Artificial Intelligence and Machine Learning, which could significantly impact our society.

Towards a pan marsupial sero-immunological tool in the demanding field of wildlife serology: Marsupial immunoglobulin-binding capability with protein A/G, protein L and anti-kangaroo antibody

by K. L. D. Tharaka D. Liyanage, Paola K. Vaz, Abdul Jabbar, Jasmin Hufschmid

Detection of infections in wildlife species is increasingly important to reduce the risk of spreading zoonotic and economically important parasites, understand disease epidemiology and promote the conservation of wildlife species. Serological tests are key in disease diagnosis and surveillance by detecting immunoglobulins against infectious agents. However, the need for species-specific reagents has limited the application of serological tests in wildlife species. This study evaluated the serum immunoglobulin-binding capability of polyclonal anti-kangaroo antibody and two non-species-specific reagents, including protein A/G and protein L, with the largest range of Australian marsupial species so far, including 32 species representing three major marsupial orders. Immunoglobulin-binding capability was assessed using immunoblotting, enzyme-linked immunosorbent assay and Western blot techniques. Variation in immunoglobulin-binding capability was observed between the three reagents and across the species tested, both across but also within taxonomic groups. Taxonomic distance was thus not always a good predictor of immunoglobulin-binding affinity, emphasizing the need to validate these reagents for each species separately. However, all three reagents bound with the serum immunoglobulins of most marsupial species tested. The findings of this study provide a valuable reference for species differences in affinity to protein A/G, protein L and anti-kangaroo antibody, assisting in the selection of appropriate reagents and the development of sero-immunological assays in Australian marsupials.

Temporal trends of hemoglobin among pregnant women: The Mutaba’ah study

by Aminu S. Abdullahi, Abubaker Suliman, Moien AB Khan, Howaida Khair, Saad Ghazal-Aswad, Iffat Elbarazi, Fatima Al-Maskari, Tom Loney, Rami H. Al-Rifai, Luai A. Ahmed

Background

Low hemoglobin (Hb) level is a leading cause of many adverse pregnancy outcomes. Patterns of changes in Hb levels during pregnancy are not well understood.

Aim

This study estimated Hb levels, described its changing patterns across gestational trimesters, and identified factors associated with these changes among pregnant women.

Materials and methods

Data from the ongoing maternal and child health cohort study–The Mutaba’ah Study, was used (N = 1,120). KML machine learning algorithm was applied to identify three distinct cluster trajectories of Hb levels between the first and the third trimesters. Descriptive statistics were used to profile the study participants. Multinomial multivariable logistic regression was employed to identify factors associated with change patterns in Hb levels.

Results

The three identified clusters–A, B and C–had, respectively, median Hb levels (g/L) of 123, 118, and 104 in the first trimester and 119, 100, and 108 in the third trimester. Cluster ’A’ maintained average normal Hb levels in both trimesters. Cluster ’B’, on average, experienced a decrease in Hb levels below the normal range during the third trimester. Cluster ’C’ showed increased Hb levels in the third trimester but remained, on average, below the normal range in both trimesters. Pregnant women with higher gravida, diabetes mellitus (type 1 or 2), nulliparity or lower level of education were more likely to be in cluster ’B’ than the normal cluster ’A’. Pregnant women who reported using iron supplements before pregnancy or those with low levels of education. were more likely to be in cluster ’C’ than the normal cluster ’A’.

Conclusion

The majority of pregnant women experienced low Hb levels during pregnancy. Changes in Hb levels during pregnancy were associated with parity, gravida, use of iron before pregnancy, and the presence of diabetes mellitus (type 1 or 2).

Determinants of anemia level among reproductive-age women in 29 Sub-Saharan African countries: A multilevel mixed-effects modelling with ordered logistic regression analysis

by Kusse Urmale Mare, Setognal Birara Aychiluhm, Kebede Gemeda Sabo, Abay Woday Tadesse, Bizunesh Fentahun Kase, Oumer Abdulkadir Ebrahim, Tsion Mulat Tebeje, Getahun Fentaw Mulaw, Beminate Lemma Seifu

Background

Despite the implementation of different nutritional and non-nutritional interventions, 43% of reproductive-age women in Africa suffer from anemia. Recent evidence also shows that none of the Sub-Saharan African (SSA) countries are on the track to achieve the nutrition target of 50% anemia reduction by 2030. To date, information on the level of anemia and its determinants among reproductive-age women at the SSA level is limited. Thus, this study aimed to estimate the pooled prevalence of anemia level and its determinants in SSA countries.

Methods

We used a pooled data of 205,627 reproductive-age women from the recent demographic and health surveys of 29 SSA countries that were conducted between 2010–2021. A multilevel mixed-effects analysis with an ordered logistic regression model was fitted to identify determinants of anemia level and the deviance value was used to select the best-fitted model. First, bivariable ordinal logistic regression analysis was done and the proportional odds assumption was checked for each explanatory variable using a Brant test. Finally, in a multivariable multilevel ordinal logistic regression model, a p-value Results

The pooled prevalence of anemia among women of reproductive age in SSA was 40.5% [95% CI = 40.2%-40.7%], where 24.8% [95% CI: 24.6%-25.0%], 11.1% [95% CI = 10.9%-11.2%], and 0.8% [95% CI = 0.7%-0.8%] had mild, moderate, and severe anemia, respectively. The prevalence significantly varied from the lowest of 13% in Rwanda to the highest of 62% in Mali, and anemia was found as a severe public health problem (prevalence of ≥ 40%) in 18 countries. The regression result revealed that polygamous marriage, women and husband illiteracy, poor household wealth, shorter birth interval, non-attendance of antenatal care, underweight, unimproved toilet and water facilities, and low community-level women literacy were positively linked with high anemia level. Additionally, the likelihood of anemia was lower in women who were overweight and used modern contraception.

Conclusions

Overall results showed that anemia among women of reproductive age is a severe public health problem in SSA countries, affecting more than four in ten women. Thus, enhancing access to maternal health services (antenatal care and contraception) and improved sanitation facilities would supplement the existing interventions targeted to reduce anemia. Moreover, strengthening women’s education and policies regulating the prohibition of polygamous marriage are important to address the operational constraints.

Blood pressure and heart rate variability responses following an acute bout of vinyasa yoga and a prolonged seated control: A randomized crossover trial

by Alexis Thrower, Bethany Barone Gibbs, Abdullah Alansare, Sally Sherman, Kelliann Davis

Vinyasa yoga is moderate-intensity physical activity, yet physiological responses are poorly characterized. The purpose of this study was to examine the effect of a vinyasa yoga session on autonomic/cardiovascular functioning in healthy adults. A randomized crossover design took place at the Physical Activity and Weight Management laboratory (Pittsburgh, PA; n = 18), and included two experimental conditions: 60 minutes of vinyasa yoga or a seated control, and measurements were taken at baseline, 5-minutes, and 65-minute post-conditions. The primary cardiovascular-related outcomes of this study included blood pressure (BP), heart rate (HR), and HR variability (HRV) measures [natural log transformed (ln) standard deviation of normal-to-normal R-R intervals (SDNN), root mean square of successive differences (RMSSD), high frequency (HF), and low frequency to high frequency ratio (LF/HF ratio)]. Linear mixed effects models were used for data analyses. Systolic BP was 8.14 mmHg lower at 5 minutes post yoga (p

Impact of mobile connectivity on students’ wellbeing: Detecting learners’ depression using machine learning algorithms

by Muntequa Imtiaz Siraji, Ahnaf Akif Rahman, Mirza Muntasir Nishat, Md Abdullah Al Mamun, Fahim Faisal, Lamim Ibtisam Khalid, Ashik Ahmed

Depression is a psychological state of mind that often influences a person in an unfavorable manner. While it can occur in people of all ages, students are especially vulnerable to it throughout their academic careers. Beginning in 2020, the COVID-19 epidemic caused major problems in people’s lives by driving them into quarantine and forcing them to be connected continually with mobile devices, such that mobile connectivity became the new norm during the pandemic and beyond. This situation is further accelerated for students as universities move towards a blended learning mode. In these circumstances, monitoring student mental health in terms of mobile and Internet connectivity is crucial for their wellbeing. This study focuses on students attending an International University of Bangladesh to investigate their mental health due to their continual use of mobile devices (e.g., smartphones, tablets, laptops etc.). A cross-sectional survey method was employed to collect data from 444 participants. Following the exploratory data analysis, eight machine learning (ML) algorithms were used to develop an automated normal-to-extreme severe depression identification and classification system. When the automated detection was incorporated with feature selection such as Chi-square test and Recursive Feature Elimination (RFE), about 3 to 5% increase in accuracy was observed by the method. Similarly, a 5 to 15% increase in accuracy has been observed when a feature extraction method such as Principal Component Analysis (PCA) was performed. Also, the SparsePCA feature extraction technique in combination with the CatBoost classifier showed the best results in terms of accuracy, F1-score, and ROC-AUC. The data analysis revealed no sign of depression in about 44% of the total participants. About 25% of students showed mild-to-moderate and 31% of students showed severe-to-extreme signs of depression. The results suggest that ML models, incorporating a proper feature engineering method can serve adequately in multi-stage depression detection among the students. This model might be utilized in other disciplines for detecting early signs of depression among people.

The lived experience of Omani adolescents and young adults with mental illness: A qualitative study

by Omar Al Omari, Gerald Amandu, Samir Al-Adawi, Zubaida Shebani, Ibtisam Al Harthy, Arwa Obeidat, Khloud Al Dameery, Mohammad Al Qadire, Iman Al Hashmi, Abduallh Al Khawldeh, Mohammed ALBashtawy, Maen Aljezawi

There is currently limited knowledge about the firsthand experiences of adolescents and young adults with mental health problems and the meanings they ascribe to these experiences, particularly within Arab countries. This study, therefore, aimed to explore the lived experience of Omani adolescents and young adults with a mental health problem. A sample of 15 participants aged 13–22 diagnosed with a range of mental health problems took part in the study. A qualitative interview guide consisting of open-ended questions was used to allow participants to speak in-depth about their experiences. Using the thematic analysis approach to uncover patterns in the data, three major themes emerged: “living in darkness”, “perilous journey” and “uncertain future”. Results show that the progress of adolescents and young adults with mental health problems is characterized by several challenges; the most significant of which is having insufficient knowledge about their illness, leading to unnecessary delays in their treatment. These findings shed light on the breadth and depth of the experience of adolescents and young adults with mental health problems and lay the groundwork for further examinations. Implications lie in the development of approaches for preventing or mitigating difficulties faced by adolescents and young adults with mental health problems.

Cost volume profit analysis for full paying patient services in Malaysia: A study protocol

by Malindawati Mohd Fadzil, Sharifa Ezat Wan Puteh, Azimatun Noor Aizuddin, Zafar Ahmed, Nor Asiah Muhamad, Abdul Aziz Harith

Dual practice within public hospitals, characterised by the concurrent provision of public and private healthcare services within public hospitals, has become a widespread phenomenon. With the participation of selected public hospitals, dual practice within public hospitals, also known as Full Paying Patient services, was an initiative the Ministry of Health Malaysia took in 2007 to retain senior specialist physicians in Malaysia. The revenue generated from the Full Paying Patient services aims to provide an avenue for public sector specialists to supplement their incomes while alleviating the Government’s burden of subsidising healthcare for financially capable individuals. However, the effectiveness of Full Paying Patient services in recouping service delivery costs and yielding a profit is still uncertain after 16 years of implementation. This study is designed to evaluate the impact of Full Paying Patient inpatient services volume, revenue, and cost on profit versus loss at selected hospitals from 2017 to 2020. From the perspective of healthcare providers, we plan to perform a cost volume profit analysis. This analysis enables us to determine the break-even point, at which total revenues match total costs, along with no-loss and no-profit thresholds for Full Paying Patient services. This study has the potential to provide insights into how variations in service volume, cost, and pricing impact healthcare providers’ profitability. It also offers critical financial information regarding the volume of services required to reach the break-even point. A comprehensive understanding of service volume, cost and pricing is imperative for making informed decisions to fulfil the objectives and ensure the sustainability of the FPP services.

How well does the virtual format of oncology multidisciplinary team meetings work? An assessment of participants’ perspectives and limitations: A scoping review

by Muhammad Abdul Rehman, Unaiza Naeem, Anooja Rani, Umm E. Salma Shabbar Banatwala, Afia Salman, Muhammad Abdullah Khalid, Areeba Ikram, Erfa Tahir

Background

Virtual multidisciplinary team meetings (VMDTM) provide a standard of care that is not limited by physical distance or social restrictions. And so, when the COVID-19 pandemic imposed irrefutable social restrictions and made in-person meetings impossible, many hospitals switched to the VMDTMs. Although the pandemic might have highlighted the ease of VMDTMs, these virtual meetings have existed over the past decade, albeit less in importance. Despite their recent importance, no review has previously assessed the feasibility of VMDTMs through the eyes of the participants, the barriers participants face, nor their comparison with the in-person format. We undertook this scoping review to map existing literature and assess the perspectives of VMDTM participants.

Material and methods

We searched MEDLINE, Embase, CINAHL, and Google Scholar from inception till July 1st, 2023 to select studies that evaluated the perspectives of participants of VMDTMs regarding the core components that make up a VMDMT. Four authors, independently, extracted data from all included studies. Two authors separated data into major themes and sub-themes.

Results

We identified six core, intrinsic aspects of a VMDTM that are essential to its structure: (1) organization, (2) case discussion and decision-making, (3) teamwork and communication, (4) training and education, (5) technology, and (6) patient-related aspect. VMDTMs have a high overall satisfaction rating amongst participants. The preference, however, is for a hybrid model of multidisciplinary teams. VMDTMs offer support to isolated physicians, help address complex cases, and offer information that may not be available elsewhere. The periodical nature of VMDTMs is appropriate for their consideration as CMEs. Adequate technology is paramount to the sustenance of the format.

Conclusion

VMDTMs are efficient and offer a multidisciplinary consensus without geographical limitations. Despite certain technical and social limitations, VMDTM participants are highly satisfied with the format, although the preference lies with a hybrid model.

Nutritional analysis and characterization of carbapenemase producing-<i>Klebsiella pneumoniae</i> resistant genes associated with bovine mastitis infected cow’s milk

by Mr. Saddam, Muddasir Khan, Muhsin Jamal, Sadeeq Ur Rahman, Abdul Qadeer, Imad Khan, Mohamed H. Mahmoud, Gaber El-Saber Batiha, Syed Hussain Shah

The current study was designed to analyze nutritional parameters and to characterize carbapenemase producing-Klebsiella pneumoniae isolates from bovine mastitic cow’s milk. Out of 700 milk samples K. pneumoniae was identified by phenotypic and molecular techniques along with their antibiogram analysis and nutritional analysis was performed using the procedure of Association of Official Analytical Chemists. Carbapenemase-producing K. pneumoniae was detected by phenotypic CarbaNP test followed by molecular characterization of their associated resistant genes blaVIM, blaKPC, blaOXA-48, blaNDM, and blaIMP along with insertion sequence common region 1 (ISCR1) and integrons (Int1, Int2, and Int3) genes. Among nutritional parameters, fat content was observed (2.99%) followed by protein (2.78%), lactose (4.32%), and total solid (11.34%), respectively. The prevalence of K. pneumoniae among bovine mastitis was found 25.71%. Antibiogram analysis revealed that more effective antibiotics was ceftazidime (80%) followed by amikacin (72%), while highly resistant antibiotics was Fusidic acid (100%). Distribution of carbapenemase producer K. pneumoniae was found 44.4%. Among carbapenem resistant genes blaKPC was found 11.25%, blaVIM 2.75%, blaNDM 17.5%, and blaOXA-48 7.5%, while blaIMP gene was not detected. Furthermore, distribution of ISCR1 was found 40%, while integron 1 was found 61.2% followed by integron 2 (20%), and integron 3 (5%). In conclusion, the recent scenario of carbapenemase resistant K. pneumoniae isolates responsible for mastitis may affect not only the current treatment regime but also possess a serious threat to public health due to its food borne transmission and zoonotic potential.
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