Seroepidemological investigation of Toxoplasma gondii and Trichinella spp. in pigs reared by tribal communities and small-holder livestock farmers in Northeastern India.

Toxoplasma gondii and Trichinella spp. are critical tissue-dwelling foodborne zoonotic parasites associated with pork consumption and pig rearing. Despite being a major pig-rearing region in the country, Northeastern India has not undergone any investigation regarding the presence of T. gondii and Trichinella spp. in pigs. Therefore, this study aims to determine the seroprevalence of T. gondii and Trichinella spp. and identify associated risk factors in pigs reared by tribal communities and small-holder livestock farmers in the northeastern region of India. In a cross-sectional serological survey, 400 pigs from 400 households across five northeastern states of India underwent testing for the seroprevalence of porcine toxoplasmosis and trichinellosis. Serum samples (80 from each state) were analyzed using commercially available ELISA assays. Data on backyard farm characteristics and various management aspects were collected, and risk factors linked with prevalence were analyzed through univariate and multivariate logistic regression analysis. The findings revealed that the apparent and true prevalence of anti-T. gondii antibodies were 45% (40.12-49.88, 95% CI) and 45.7% (40.7-50.69, 95% CI), respectively. As for anti- Trichinella antibodies, both the apparent and true prevalence were 0.75% (-0.1-1.6, 95% CI). The univariate and multivariate analyses indicated that age above 24 months (OR 7.20, 95% CI 2.45-23.71), exposure to cats (OR = 5.87, 95% CI 2.55-14.05), and farms operating for breeding purposes (OR = 5.60, 95% CI 3.01-11.04) were significant risk factors associated with the seroprevalence of T. gondii. This study marks the initial documentation of the seroprevalence of T. gondii and Trichinella spp. in pigs reared by tribal communities in Northeastern India. The results emphasize the significance of these parasites as foodborne zoonotic threats in the region, potentially posing substantial public health risks, especially within tribal and rural communities. The insights derived from this research could be valuable in formulating targeted preventive and control strategies against T. gondii and Trichinella spp. in pigs, not only in this region but also in areas with similar rearing practices.

First Seroepidemiological Investigation of Hepatitis E Virus Infection in Backyard Pigs from Northeastern India: Prevalence and Associated Risk Factors.

Hepatitis E virus (HEV) is the leading cause of acute viral hepatitis globally, with zoonotic potential, and pigs are considered the major reservoir. To determine the seroprevalence of HEV infection in pigs reared in backyard conditions in the northeastern region of India, blood samples were collected from 400 pigs from five northeastern states (80 samples from each state) and tested for IgG antibodies against HEV using an ELISA assay. Questionnaires on farm characteristics and management practices were completed, and risk factors associated with HEV were studied using univariate and multivariate analysis. The apparent seroprevalence of HEV infection was 51% (46.1-55.9, 95% CI), with a true prevalence of 52.98% (47.22-58.75, 95% CI). The risk factors significantly associated with higher HEV seropositivity were as follows: lack of disinfection (OR 4.65), feeding swill (restaurant and bakery waste) (OR 2.55), failure to follow the all-in-all-out production system (OR 3.47), and medium holding size (OR 9.83), which refers to mixed rearing of younger and older age groups. This study demonstrates that HEV is widespread among pigs reared in northeastern India. The risk factor analysis conducted in this study provides valuable insights into the prevalence of HEV in the region.

Morphometry of depth of interhemispheric fissure on Indian cadaveric brain specimens.

Background: During neurosurgeries like resection of interhemispheric lipoma or cyst, surgeon needs to remain within the limits of interhemispheric fissure (IHF). Despite a massive literature search, data regarding the morphometry of IHF is meagre. Therefore, the present study was done to calculate the depth of IHF. Materials and Methods: Twenty-five (fourteen male and eleven female) fresh human cadaveric brain specimens were used. The depth of IHF was measured from frontal pole; three points, anterior to coronal suture (A, B and C); four points, posterior to coronal suture (D, E, F and G) and from two points (via parieto-occipital sulcus and calcarine sulcus) on occipital pole. The measurements were taken from these points up to the floor of IHF. IHF is a midline groove and hence the measurements were taken from each point against both the left and the right cerebral hemispheres. At the end, not much bilateral asymmetry was found, hence the average of the reading for the same point against left and the right cerebral hemisphere was considered for calculation. Result: Maximum depth was found to be 59.60 mm and minimum depth was found to be 19.66 mm among all the points which were considered for evaluation. No statistical difference was found in the depth of IHF among the male and the female groups as well as in the various age groups. Conclusion: This data and knowledge about the depth of interhemispheric fissure will aid the neurosurgeons in order to perform the interhemispheric transcallosal approach as well as surgeries of interhemispheric fissure such as excision of lipoma, cyst, tumor of interhemispheric fissure through the shortest and the safest possible route.

Morphometry of Anterior Cruciate Ligament in Knee joint. A Cadaveric Study.

Objectives: The journey of Anterior Cruciate Ligament (ACL) reconstruction surgery has long way from open surgery to arthroscopy. Therefore, the anatomical features of the ACL and its two bundles description rewarded as good outcome of procedure. Material and methods: Fifty fresh knees were dissected. The insertion position, length and diameter of ACL and patellar tendon length were measured and determines the range of normality according to sex and side. Results: The average lengths of Antero- medial (AM) and postero- lateral (PL) bundle of the ACL were 35.35 mm and 26.11mm in male on right side, respectively while 34.21mm and 25.53 mm in female. Patellar tendon length was 45.24 mm in right side of male and 43.38 mm in female obtained. The average lengths of AM and PL bundle of the ACL were 35.47 mm and 26.04 mm in male on left side, respectively while 34.23 mm and 25.38 mm in female. Conclusion: The length of ligaments was higher in male than female. There was strong correlation between height and length of ligaments but no correlation with age.

Anatomical Variations of Corona Mortis in the Anterior Intrapelvic Approach: A Cadaveric Study.

Corona mortis (CMOR) is an anastomotic channel either arterial, venous or both connecting the obturator and external iliac systems excluding aberrant channels in the retropubic space. The goal of this study is to illustrate the type of CMOR via the Anterior Intrapelvic approach (AIP) which has not been studied in tandem. This descriptive observational study was performed in the Department of Anatomy, All India Institute of Medical Sciences, Jodhpur, Rajasthan, India from January 2018 to December 2020. It is a potential culprit for significant haemorrhage in scenarios of pelvic trauma or when sectioned accidentally, augmenting difficult haemostasis behind the superior pubic ramus. Previously reported to be exclusively arterial, the venous variant has now emerged as the more frequent anastomosis. AIP was performed on 31 adult human cadavers of 62 hemipelvises to record variations and statistical analysis of retropubic anastomosis with respect to gender, weight groups and side distribution was done. Corona Mortis was observed in 50(80.6%) hemipelvises. To ramify, venous CMOR and arterial CMOR were exclusively encountered in 25(40.3%) and 10(16.10%) hemipelvises respectively. Males recorded a higher prevalence of CMOR. Despite being a frequent anastomotic variant, it is a rare entity in textbooks. CMOR can potentially alter patient outcome and have serious implications in pelvic surgeries via AIP owing to its easier accessibility. Besides, venous CMOR being more common, a venous bleed is inherently more arduous to manage in the pelvis. A larger calibre, less frequent aberrant channel could also prove to be lethal in its injury. Hence, cadaveric simulation of the technique is paramount to master the technique and to mitigate catastrophic vascular events.

Tibio-femoral Angle Assessment and Its Radiological Co-relation in Healthy Rajasthani Population.

Existence of variations in morphological proportions in human beings has led to the development of different values of tibio femoral angle (TFA) in different populations which is undoubtedly useful in clinical orthopaedic surgery. Meagre literature is available among Indian population. The cross-sectional study was creating baseline data of TFA among Rajasthani population of India with reasonable accuracy and also to correlate these data to radiological findings during July 2014-June 2016. The TFA (in degrees) of 500 healthy subjects from various regions of Rajasthan were measured in standing and supine positions. In addition, the TFA of 134 persons (randomly selected) from study group were also measured on roentgenogram. Subsequently both of these data were compared. The normal range of TFA angle in Rajasthani population was in between 160°-174°. TFA in standing position on right side was 170.96° (with 95% Confidence Interval (CI) = 170.24 to 171.68) and on left side was 170.14° (with 95% CI = 169.21 to 171.06). Mean TFA in supine position on right side in age group from 21 to 30 years was 167.12±6.73° and in age group from 31-40 years was 167.668±6.7°. No significant difference was observed in posture change and according to age. Statistically significant difference was observed in between radiological and gross measurement which was more in radiological than to clinical measurement of TFA (mean difference = +5.78° units) with 95% CI for the difference is (4.03°-7.53°). Higher values of tibio-femoral angle (TFA) were observed in males as compared to females and in radiological measurements.

Peroxisome proliferator-activated receptor-γ signalling protects hair follicle stem cells from chemotherapy-induced apoptosis and epithelial-mesenchymal transition.

BACKGROUND: Permanent chemotherapy-induced alopecia (pCIA), for which preventive interventions remain limited, can manifest with scarring. While the underlying pathomechanisms of pCIA are unclear, depletion of epithelial hair follicle (HF) stem cells (eHFSCs) is likely to play a role. OBJECTIVES: To explore the hypothesis that, besides apoptosis, eHFSCs undergo pathological epithelial-mesenchymal transition (EMT) in pCIA, thus explaining the scarring phenotype. Furthermore, we tested whether a peroxisome proliferator-activated receptor (PPAR)-γ modulator could prevent pCIA-associated pathomechanisms. METHODS: Organ-cultured human scalp HFs were treated with the cyclophosphamide metabolite 4-hydroperoxycyclophosphamide (4-HC). Additionally, HFs were pretreated with the agonistic PPAR-γ modulator N-acetyl-GED-0507-34-Levo (NAGED), which has previously been shown to promote K15 expression and antagonize EMT in eHFSCs. RESULTS: In accordance with anticipated hair bulb cytotoxicity, dystrophy and catagen induction, 4-HC promoted apoptosis along with increased p53 expression, DNA damage and pathological EMT in keratin 15+ (K15) eHFSCs, as evidenced by decreased E-cadherin expression and the appearance of fibronectin+ and vimentin+ cells in the hair bulge. Pretreatment with NAGED protected against 4-HC-induced hair bulb cytotoxicity/dystrophy, and apoptosis, p53 upregulation and EMT in the bulge, thereby significantly preventing depletion of K15+ human eHFSCs ex vivo. CONCLUSIONS: Since a key cyclophosphamide metabolite alone suffices to damage and deplete human scalp eHFSCs by promoting apoptosis, DNA damage and EMT ex vivo, strategies to prevent pCIA need to target these pathomechanisms. Given the ability of NAGED to prevent chemotherapy-induced eHFSCs damage ex vivo, our study introduces the stimulation of PPAR-γ signalling as a novel intervention strategy for the prevention of pCIA.

Carrying Angle in Population of Western India: Normative Values.

Carrying angle is one of the secondary sexual characteristics which is important for keeping the hand away from body to improve functionality. A cross-sectional survey was conducted in Western Indian population (rural and urban population in and around Jodhpur district) during July 2014-June 2016 with the purpose of evaluating the carrying angle in apparently healthy adult and determines the range of normality according to age, sex, height, waist circumference and special reference to handedness. The carrying angle (elbow) was measured in 504 adults aged between 20-40 years (by simple random sampling) by a digital Goniometer. The subjects were in anatomical position (stand up straight, roll his/her shoulders back and the elbow was fully extended and the forearm fully supinated) while the carrying angle were taken. Carrying angle of the right side in male's 11.54°±1.29° and in female's 13.41°±1.26°, whereas these value in left side 10.21°±1.34° and 12.19°±1.44° respectively. All measurements in females are significantly higher in comparison to males (p<0.001). The significant difference was observed in carrying angle according to handedness; dominant hand had higher value of carrying angle in comparison to non-dominant hand in both gender (p<0.001). The height of individuals is positively correlated with the angle whereas there was no correlation was found in between age, waist circumference. The present study showed that the females have greater carrying angle than males.

Estimation of Fertilization Rate, Implantation Rate, Pregnancy Rate and Live Birth Rate in Intracytoplasmic Sperm Injection in Association with Number of Attempts and Sperm Morphology.

Infertility is a growing health problem. It affects relatively large number of couples both globally as well as in India. Fertility treatment registers suggest that infertility is on increase. With the advancement in the knowledge and technology about the In Vitro fertilization (IVF) process, In Vitro fertilization success rates are improving over the time. In many cases of male-factor infertility, Intracytoplasmic sperm injection (ICSI) is the recommended treatment. The aim of this study was to estimate fertilization, implantation and pregnancy rates in ICSI cycles in association with number of attempts and sperm morphology. A total of 250 IVF-ICSI cycles were performed at a Fertility Center in Jaipur, Rajasthan, India. This cohort study was performed during 2013 to 2015. The differences in the outcomes of ICSI in association with sperm morphology and number of ICSI attempt were evaluated. Significantly higher mean was observed in fertilization rate and live birth rate with ≥5% sperm morphology 79.63% vs. 71.46% and 20% vs. 13.9% respectively). Significantly higher mean implantation rate and clinical pregnancy rate (CPR) and live birth rate were observed with attempt 3 as compared to 1 and 2 attempts. IVF success rates are improving over the time as knowledge and technology are advancing. Sperm morphology and number of attempt affect the ICSI outcomes.

Reference Charts of Fetal Biometric Parameters at Different Gestational Age Groups in Indian Population.

To develop, a reference charts of fetal abdominal circumference and femur length in normal pregnant women by using ultrasonography for Indian population. Further comparison of our findings with data derived from different population. This retrospective cross-sectional study included 300 normal singleton pregnancies and was carried out in the department of Anatomy, All India Institute of Medical Sciences, Jodhpur (Rajasthan), India from September 2017 to April 2019. Ultrasonography measurements included Abdominal circumference (AC) and Femur length (FL). Reference charts with mean AC and FL for corresponding Gestational age (GA) in weeks were developed. Also reference centiles (10th, 50th, 90th and 95th) were derived from this model and compared with similar studies done on different population. There was no statistically significant difference in age distribution of pregnant women (p=0.87). Both AC and FL were found to have statistically significant linear relationship with advancing gestational age (p=0.0005 & p=0.0003 respectively). There is significant difference observed between the values obtained in present study with studies concluded on Chinese and European population. Measurement of AC and FL are used to predict gestational age of fetus by using various regression formulae, also AC is known to be good predictor of nutritional status of fetus in utero. A separate chart is required for every different population because ethnicity, nutrition and environmental factor can have impact on normal values. Therefore, a reference chart for these parameters according to Indian population standards is essential to avoid misinterpretation of data. This would help to avoid misdiagnosis of intrauterine growth retardation or macrosomia during prenatal and perinatal period.

Development of novel visual detection methodology for Salmonella in meat using saltatory rolling circle amplification.

AIM: The aim of this study was to develop a saltatory rolling circle amplification (SRCA) assay for rapid, simple and visual detection of Salmonella in meat. METHODS AND RESULTS: Saltatory rolling circle amplification assay was established using simple PCR primers targeting the invA gene of Salmonella enterica. The specificity of the SRCA assay was determined using 28 Salmonella and 15 non-Salmonella strains. The analytical sensitivity of the developed SRCA, conventional and real-time PCR assays were 70 fg, 7 pg and 700 fg S. enterica DNA per tube, respectively. The limit of detection (LoD) of the SRCA assay was 40 CFU per gram of meat without enrichment and 4 CFU per gram after including 6 h brief enrichment step. The detection limits of 40 CFU per gram and 4 CFU per gram of meat were achieved within 165 min and 9 h, respectively (including DNA extraction). To assess the real-world relevance of the SRCA assay, it was used to screen Salmonella from the field pork samples (n = 82). The same samples were also tested with culture (ISO 6579: 2002) method, conventional and real-time PCR assays. Using the developed assay with 6-h enrichment step, it could give accurate results as that of the culture method. CONCLUSIONS: The results of this study showed that the SRCA assay is a rapid, simple, sophisticated equipment-free and user-friendly method for accurate detection of Salmonella in meat foods. To our information, this is the first study to deploy SRCA assay for screening foods for Salmonella. SIGNIFICANCE AND IMPACT OF THE STUDY: The developed SRCA assay is cost-effective, easy-to-perform and equipment-free; therefore, it has the potential to replace other molecular detection methods for regular screening of Salmonella in foods in field laboratories.

Enhancing Student Centered Active-Learning by "Flipped Classrooms" In a Histology Module.

Flipped learning has been the focus of significance and area of interest for medical educationists but accounts of the progression and the assessment of this pedagogical prototype are lacking. The present study describes how a histology module was delivered using the flipped learning approach. This qualitative and quantitative analysis was undertaken among First Professional MBBS students studying human anatomy at All India Institute of Medical Sciences, Jodhpur Rajasthan, India from 2016 to 2019. The participants were subjected to conventional and active learning approach i.e. flipped classroom for the various separate topics of histology with comparable level of difficulty. The students were assessed for subject knowledge by a short assessment which was taken after each session. A feedback from them in reference to use of flipped learning approach in regular teaching-learning and their experience was also obtained. Both the conventional and flipped classroom teaching showed significant changes in knowledge of participants, though mean test score was higher in the flipped learning activity. In the feedback analysis it was noted that majority of the medical students considered incorporation of active learning methods useful for learning histology. Use of active teaching-learning by flipped classroom can assist as a comprehensive tool in regular histology sessions.

Estimation of Fetal Weight: An Ultrasonography Study in Indian Population.

To create a reference chart for estimated fetal weight (EFW) in normal pregnancy for use in Indian population and compare it with reference chart from other population. This retrospective cross-sectional study included 300 normal singleton pregnancies coming for routine antenatal ultrasonography examination and was carried out at All India Institute of Medical Sciences, Jodhpur from September 2017 to April 2019. Ultrasonographic measurements included fetal biparietal diameter (cm), head circumference (cm), abdominal circumference (cm) and femur length (cm). Estimated fetal weight was calculated by using Hadlock algorithm which is already fed into ultrasonography machine. Reference chart with mean EFW for corresponding Gestational age (GA) in weeks was developed. Also Reference centiles (10th, 50th, 90th and 95th) were derived from this model. There was no statistically significant difference in age distribution of pregnant women (p=0.87). Statistically significant linear relationship found between EFW and advancing gestational age (p=0.0004). Maximum gain in EFW (34.05%) was observed after second trimester (28 week). Maximum and minimum fetal weight at 38 weeks of GA in our study found to be 3389 grams and 2567 grams respectively, which has significant difference. This could be due to huge difference in socio-economic and nutritional status among Indian population which might have impacted on maternal and fetal health. Estimated fetal weight was found to be at lower range in Indian population compared to reference chart developed into western population. Fetal weight to their corresponding GA is an important factor in determining growth and development in fetus. In normally developing fetus the EFW has linear correlation with advancing GA. A separate reference chart is required for every different population because ethnicity, nutrition and environmental factor can have impact on normal EFW values. This would help to avoid misdiagnosis of intrauterine growth retardation or macrosomia in fetuses and hence unnecessary medical interventions can be prevented during prenatal and perinatal period.

Morphometric evaluation of Anterior Cerebral Artery (on Digital Subtraction Angiography)-Potential implications.

PURPOSE: The aim of the present study was to perform a morphometric analysis of ACA and to establish significant differences, if any, with sex and age via Digital Subtraction Angiography (DSA). METHODS: This cross-sectional study was conducted on 70 patients (>20 years) for the evaluation of ACA by Digital Subtraction Angiography done on a Philips biplane system clarity (Allura FD20/20). Length and diameter of ACA were taken manually on the DSA console using auto-calibration. Statistical analysis was done. RESULT: Mean diameter and length of ACA was found to be greater in female. The f-ratio value for diameter and length of the same was 1.43 and 0.92 respectively. The length of ACA was found to be statistically significant for different age groups. The mean length of ACA followed a decreasing trend with age, whereas mean diameter of ACA was nearly the same in all the age groups. CONCLUSION: The results of our study show that the length of ACA was found to be statistically significant for different age groups.

Identification of a novel cluster of PCV2 isolates from Meghalaya, India indicates possible recombination along with changes in capsid protein.

Documentation of the emergence of Porcine circovirus 2 (PCV2) infection and economic losses incurred due to high mortality has been reported worldwide. The prevalence and genetic diversity of the virus has been reported in Northeast India including the possible chances of Classical swine fever virus (CSFV) vaccine failure in pig population in this region resulting in major disease outbreak. Irrespective of the genetic variability, the emergence of a novel cluster (based on the ORF2 phylogeny) was reported last year. The present study describes a state-wide (Meghalaya, India) molecular epidemiological investigation of PCV2 strains in pig population by amplification, sequencing and undertaking phylogenetic analyses. The results indicate the identification of a novel cluster of PCV2 originating from the inter-genotypic recombination between PCV2c and PCV2d. Multiple sequence alignment of amino acids indicates possible substitution in the A, B and C domains of the capsid protein. Molecular structural modelling of the capsid protein of PCV2 indicated possible motif variations in the secondary structure including presence of a tunnel, encountered at the interface region on each chain facilitating in transportation of molecules and acting as an active site for attachment and penetration. The baseline data strengthens the existing control programme of PCV2 and is possibly helpful in the planning of active surveillance strategy in this region.

Detection of classical swine fever virus E2 gene in cattle serum samples from cattle herds of Meghalaya.

The present study focused on the detection and genetic characterisation of 5' untranslated region (5'UTR) and E2 gene of classical swine fever virus (CSFV, family Flaviviridae, genus Pestivirus) from bovine population of the northeastern region of India. A total of 134 cattle serum samples were collected from organised cattle farms and were screened for CSFV antigen with a commercial antigen capture enzyme linked immunosorbent assay (Ag-ELISA) and reverse transcription-polymerase chain reaction (RT-PCR). A total of 10 samples were positive for CSFV antigen by ELISA, while all of them were positive in PCR for 5'UTR region. Full length E2 region of CSFV were successfully amplified from two positive samples and used for subsequent phylogenetic analysis and determination of protein 3D structure which showed similarity with reported CSFV isolate from Assam of sub-genogroup 2.1, with minor variations in protein structure.

Seroprevalence of peste des petits ruminants and bluetongue in goat population of Meghalaya, India.

AIM: This study aimed to determine the seroprevalence of peste des petits ruminants (PPR) and bluetongue (BT) in goats' population in the state of Meghalaya of Northeast India. MATERIALS AND METHODS: The serosurveillance study was done from the random sampling (n=598) of blood collected from five districts (Ri-Bhoi, East Khasi Hills, West Khasi Hills, Jaintia Hills and West Garo Hills) of Meghalaya. The presence of antibodies against PPR and BT in the samples was detected by indirect enzyme-linked immunosorbent assay (ELISA) method for PPR and competitive ELISA for BT. RESULTS: The results showed the overall seropositivity of PPR and BT at 7.19% and 60.20%, respectively. West Garo Hills recorded the highest seroprevalence of both PPR (9.81%) and BT (68%) and 3.6% of the samples tested positive for both PPR and BT. CONCLUSION: The random survey results indicating the presence of PPR and BT have specific implication in epidemiological perspectives since it highlights the prevalence under natural situations, where the subclinical, inapparent, or non-lethal or recovery of infection was suspected in unvaccinated animals. It also warrants further studies to suggest appropriate control measures to prevent the spread of infection.

Detection of Peste des petits ruminants virus and goatpox virus from an outbreak in goats with high mortality in Meghalaya state, India.

AIM: We describe a laboratory investigation carried out to confirm the etiology of the heavy mortality (37 animals died out of total 44, i.e. 84%) in goats in Ri-Bhoi district of Meghalaya, Northeast region of India in December 2015. The clinical signs observed were abortion, diarrhea, high fever (up to 104°F), pox lesion in the skin, and respiratory distress. MATERIALS AND METHODS: The samples comprising whole blood, sera, and pox lesion were collected from the animals (n=7) from an outbreak for the screening of peste des petits ruminants (PPR) and poxviruses. The whole blood and sera were used for screening of PPR virus (PPRV) by sandwich enzyme-linked immunosorbent assay (ELISA) and antibody by competitive ELISA as well as detection of PPRV partial N gene by reverse transcription-polymerase chain reaction (PCR). The skin lesions were used for the detection of poxvirus by PCR. RESULTS: The results showed the presence of PPR antigens (58-80%) in the samples by sandwich ELISA and antibody in all the sera samples ranging from 9% to 41% positivity in competitive ELISA. Four samples were positive for PPRV partial N gene. The skin lesion screened for poxvirus was also found to be positive for I3L gene of goatpox virus. CONCLUSION: We confirm the outbreak of disease in goats with high mortality is a case of mixed infection of PPR and goatpox detected for the first time in Northeast India.

Molecular Characterization and Computational Modelling of New Delhi Metallo-ß-Lactamase-5 from an Escherichia coli Isolate (KOEC3) of Bovine Origin.

Emergence of antimicrobial resistance mediated through New Delhi metallo-ß-lactamases (NDMs) is a serious therapeutic challenge. Till date, 16 different NDMs have been described. In this study, we report the molecular and structural characteristics of NDM-5 isolated from an Escherichia coli isolate (KOEC3) of bovine origin. Using PCR amplification, cloning and sequencing of full blaNDM gene, we identified the NDM type as NDM-5. Cloning of full gene in E. coli DH5α and subsequent assessment of antibiotic susceptibility of the transformed cells indicated possible role of native promoter in expression blaNDM-5. Translated amino acid sequence had two substitutions (Val88Leu and Met154Leu) compared to NDM-1. Theoretically deduced isoelectric pH of NDM-5 was 5.88 and instability index was 36.99, indicating a stable protein. From the amino acids sequence, a 3D model of the protein was computed. Analysis of the protein structure elucidated zinc coordination and also revealed a large binding cleft and flexible nature of the protein, which might be the reason for broad substrate range. Docking experiments revealed plausible binding poses for five carbapenem drugs in the vicinity of metal ions. In conclusion, results provided possible explanation for wide range of antibiotics catalyzed by NDM-5 and likely interaction modes with five carbapenem drugs.