Microbial Profiling, Antimicrobial Resistance Surveillance, and Molecular Detection of MecA Gene in Staphylococcal Strains from Donor Human Milk: Insights from a Milk Bank Investigation.

PURPOSE: Donor human milk (DHM) from milk banks provides vital nutrition to vulnerable infants. Understanding its microbial profile and antimicrobial resistance patterns is crucial for ensuring its safety and efficacy. This study aimed to profile the microbial composition, detect antibiotic resistance, and identify the presence of mecA gene in Staphylococcal strains from DHM samples. MATERIALS AND METHOD: A total of 151 DHM samples were collected from a regional human milk bank in North India. Microbial identification was performed using MALDI TOF MS, and antimicrobial susceptibility testing was conducted using the disc diffusion method. Molecular methods, including PCR, were employed for mecA gene detection. RESULTS: The study revealed a diverse microbial profile, with Staphylococcus species being predominant. Acinetobacter and Pseudomonas species were also prevalent, raising concerns due to their association with healthcare-associated outbreaks. High rates of antibiotic resistance were observed across both Gram-positive and Gram-negative bacteria, with resistance to commonly used antibiotics such as penicillin, clindamycin, erythromycin, and ceftriaxone. The mecA gene, associated with methicillin resistance, was detected in a significant proportion of Staphylococcal isolates. CONCLUSION: The study underscores the importance of rigorous microbial analysis and antimicrobial susceptibility testing in assessing the safety of DHM. The presence of diverse microbial species, including antibiotic-resistant strains and the mecA gene in Staphylococcal strains, emphasizes the need for stringent hygiene practices and continuous surveillance in milk banks. Implementing comprehensive screening protocols and adhering to best practices in milk handling and pasteurization are crucial for safeguarding the health of vulnerable infants reliant on donor milk.

Postexchange transfusion-related acute lung injury in a term neonate.

We report a successful case where a newborn with transfusion-related acute lung injury following an exchange transfusion was effectively treated using conservative methods, eliminating the need for surfactant therapy. Very few instances of this complication have been documented globally. A low birth weight, small for gestational age, term neonate, diagnosed with hyperbilirubinaemia due to Rh incompatibility, experienced sudden respiratory distress in the form of severe retractions, tachypnoea and cyanosis 3 hours after the procedure. Neonate required mechanical ventilation on the grounds of mixed acidosis and diffuse alveolar infiltrates on the chest radiograph. The medical team suspected and treated the baby for transfusion-related acute lung injury through conservative measures. Transfusion-related acute lung injury, an acute life-threatening complication of blood component transfusion, can exhibit symptoms in neonates that are frequently misinterpreted as sepsis. The baby was discharged in good health after successful management after 19 days.

Pyridoxine-Dependent Epilepsy With Poor Neurodevelopmental Outcome: Case Report.

Pyridoxine-dependent seizures are a rare cause of recurrent seizures in the neonatal period that are resistant to most of the antiepileptic medications, but respond to pyridoxine. There is a wide spectrum of clinical manifestations, and in the absence of biochemical markers, clinical diagnosis is often delayed. We report a case of neonatal seizures that initially responded to antiepileptic drugs but later presented with intractable seizures and metabolic abnormalities. Clinical exome sequencing was suggestive of ALDH7A1 mutation.

Congenital myopathy presenting as recurrent pneumonia with lung collapse and pulmonary artery hypertension.

A boy presented with cough, breathlessness for 1 month, fever for 1 week with similar previous episodes without hospitalisation. He had generalised muscle wasting, acute chronic malnutrition and required immediate ventilation. Provisional diagnosis of recurrent pneumonia with failure to thrive was made. As serial chest X-rays showed recurrent lung collapse, congenital lung anomalies were ruled out. 2D-echocardiography showed pulmonary arterial hypertension. Workup for congenital immunodeficiency and cystic fibrosis was negative. There was no improvement in muscle mass despite total parenteral nutrition. He was noticed to have myopathic facies. History was reviewed when the mother reported reduced fetal movements in this pregnancy. The patient had low voice amplitude. Creatine kinase levels were normal. Muscle biopsy followed by whole exome sequencing identified frameshift duplication NM_020451.3(SELENON):c.249_250dupGG (p.Asp84Glyfs*17), thus, confirming diagnosis of SEPN1-related congenital myopathy (CM) with fibre-type disproportion. Respiratory system involvement was distracter, emphasising consideration of CM while evaluating persistent lung collapse with muscle wasting.

Arthrogryposis, renal dysfunction, cholestasis syndrome in a neonate: an uncommon association of common problems.

A male infant born out of non-consanguineous marriage to a primigravida presented to us as his third hospitalisation with ichthyotic lesions all over the body, cholestatic jaundice, multiple joint contractures and a history of recurrent sepsis. Blood and urine investigations revealed Fanconi syndrome, hypothyroidism and direct hyperbilirubinaemia with elevated liver enzymes and normal gamma glutamyl transpeptidase levels. The combination of arthrogryposis, renal dysfunction and cholestasis led to the suspicion of arthrogryposis, renal tubular dysfunction, cholestasis (ARC) syndrome, which was then proved by genetic testing. The baby was managed conservatively with respiratory support, antibiotics, multivitamins, levothyroxine and other supportive measures but succumbed to the illness on day 15 of hospitalisation. Genetic analysis using next-generation sequencing was confirmatory of a homozygous mutation in VIPAS39 gene leading to ARC syndrome type 2 in the present case. Genetic counselling was provided and prenatal testing was advised to the parents for future pregnancies.

Role of cord blood IGF-1 and maternal HbA1c levels to predict interventricular septal hypertrophy among infants of diabetic mothers: A case-control study.

BACKGROUND: Infants of diabetic mothers (IDMs) develop interventricular septal hypertrophy (ISH) (> 6 mm) [1]. The proportion of IDMs developing ISH varies from country to country. Maternal HbA1c and cord blood Insulin-like growth factor-1 (IGF-1) levels have been found useful to predict ISH. METHODS: This was a case-control study of term neonates of diabetic mothers (cases) and of non-diabetic mothers (controls) to evaluate echocardiographic (ECHO) differences among cases and controls and to find the correlation of interventricular septal thickness (IVS) thickness with maternal HbA1C and cord blood IGF-1 levels. RESULTS: Of 32 cases and 34 controls (mean gestational age 37.7 ± 0.9 weeks), 15 (46.8 %) cases, no control developed ISH. Septal thickness was more (6 ± 0.15 cm vs 3 ± 0.06 cm; p = 0.027) in cases than controls. Functional ECHO parameters including left ventricle ejection fraction were comparable (p = 0.9) among the two groups. Maternal HbA1C levels were higher (6.5 % ± 1.3 vs 3.6 % ± 0.7; p = 0.001) with a positive correlation with IVS (Pearson's coefficient 0.784, p < 0.001). Cord blood IGF1 levels were too higher in cases (99.1 ± 6.09 ng/ml vs 37.1 ± 2.99 ng/ml; p < 0.001) with moderate correlation with IVS thickness (Pearson's coefficient 0.402; p = 0.00). Receiver operator curve analysis showed, that at a cut-off of 72 ng/ml, cord blood IGF1 predicted ISH with 72 % sensitivity; 88 % specificity and at a cut-off of 7.35 %, maternal HbA1c predicted ISH with sensitivity; specificity of 93.8 % and 72.1 % respectively. CONCLUSION: ISH was present in 46.8 % in cases as compared to none in controls. IVS thickness correlated well with maternal HbA1C and moderately with cord blood IGF-1 levels. Functional parameters on ECHO were unaffected by maternal diabetic control. At the cut-off of maternal HbA1c of 7.35 % and cord blood IGF-1 of 72 ng /ml, babies need to be monitored clinically with ECHO to look for ISH.

Breastfeeding rates at discharge for very low birthweight neonates and their determinants: An observational study from a tertiary care neonatal intensive care unit in India.

BACKGROUND: Mother's milk is the best milk for neonates. Preterm very low birthweight (VLBW) neonates face many challenges leading to low rates of breastfeeding at discharge. AIMS: (i) To determine the proportion of <32 weeks preterm VLBW neonates who are exclusively breastfed (EB) at discharge and (ii) determinants of exclusive breastfeeding (EBF) at discharge. METHODS: An observational study was conducted for a duration of 1 year, from May 2019 to April 2020 in a tertiary care neonatal intensive care unit (NICU) in North India. Consecutive <32 weeks preterm VLBW neonates admitted within 72 h of birth and on full enteral feeds (FEF) within 10 days of birth were included in this study and followed up till discharge. RESULTS: Forty-four of 97 (45.4%) preterm VLBW neonates were exclusively breastfed and 31/97 (32%) received more than 80% mother's own milk (MOM) at discharge. Male sex (P = 0.03), those whose first feed had any amount of MOM (P = 0.038) or exclusive MOM in their first feed or when initiated on first FEF (P = 0.002), and neonates with longer duration of hospital stay (P = 0.035) had an increased chance of being exclusively breastfed at discharge. CONCLUSION: Preterm VLBW neonates who receive any amount of MOM in their first feed or first FEF, male infants and those who stay longer in hospital are more likely to be exclusively breastfed at discharge.

Oral Application of Mother's Own Milk for Prevention of Late Onset Sepsis in Preterm Very Low Birth Weight Neonates: A Randomized Controlled Trial.

Background: Oral application of mother's own milk (MOM) in very low birth weight (VLBW) neonates may provide an effective method of cleaning the oral cavity and influencing the immunocompetent cells there. Methods: A double-blind randomized controlled trial was conducted in a tertiary level neonatal unit on VLBW neonates; that is, neonates with birth weight <1,500 g, hospitalized within 72 hours of birth. The intervention was oral care with MOM. The primary objective was to observe the effect of oral care with MOM on late onset sepsis (LOS) and also on mortality, days to attain full enteral feed, necrotizing enterocolitis (NEC), exclusive breastfeeding rates at discharge, and total hospital stay (THS). Results: One hundred ten neonates were randomized to intervention and no-intervention group; 55 in each group. Neonates in the intervention group developed LOS in 9.1% versus 27.3% in no-intervention group (p = 0.013). Mortality was 9.1% versus 16.4% (p = 0.252); days to attain full enteral feeds were 6.94 ± 2.34 versus7.4 ± 2.31 days (p = 0.306); exclusive BF rates at discharge were 83% versus 75% (p = 0.449); THS was 30.72 ± 8.72 versus 33.5 ± 8.7 days (p = 0.267) in the two groups, respectively. There was no effect on NEC. Conclusion: Oral care of VLBW neonates with MOM reduces LOS and mortality.

Vitamin D Status of Very Low Birth Weight Neonates at Baseline and Follow-up after Daily Intake of 800 IU Vitamin D.

INTRODUCTION: Vitamin D deficiency (VDD) is rampant in neonates. Recommendations for supplementation are variable. METHODS: An observational study was done on less than 32 weeks of very low birth weight neonates to find prevalence of VDD (<20 ng/ml) at baseline; at 38 ± 2 weeks post-menstrual age (PMA) after daily intake of 800 IU vitamin D (vit D). Secondary objectives were to find determinants of VDD, to compare growth in deficient; vit D sufficient (VDS) neonates; to find vit D toxicity. RESULTS: Of 83 neonates, 81 (97.6%) were VDD at baseline and 5 (6%) at 38 ± 2 weeks PMA. Determinants for VDD at baseline were inadequate maternal sun exposure (p < 0.001) and vit D supplementation (p = 0.007). Factors for VDD at 38 ± 2 weeks PMA were male gender (p = 0.049), morbidities (p = 0.006), ventilation >24 h (<0.001), sepsis (p = 0.032), caffeine (p ≤ 0.001) and missed supplements (p < 0.001). Weight and length gain of VDD to VDS neonates were (6.70 ± 2.40 to 8.96 ± 2.21 g/day); (0.82 ± 0.34 to 1.08 ± 0.37 cm/week), respectively (p < 0.001). Head circumference gain (cm/week) of VDS; VDD neonates was 0.58 ± 0.09; 0.54 ± 0.06 (p = 0.054), respectively. No neonates developed vit D toxicity. CONCLUSIONS: In preterm VLBW neonates, the prevalence of VDD was 97.6% but decreased by >90% at 38 ± 2 weeks with a daily intake of 800 IU vit D. Inadequate maternal vit D intake and sun exposure determined low baseline vit D status of neonates. Male gender, morbidities, ventilation, sepsis, caffeine, missed vitamin D supplements were determinants of poor vit D status at follow-up. Weight gain and length increments were more in the VDS group.

Molecular docking and dynamic studies of crepiside E beta glucopyranoside as an inhibitor of snake venom PLA2.

Alternative treatments from plant-derived small molecules for neutralizing the venom lethality in snake envenomation are prevalent now. Elephantopus scaber, a tropical plant species has been recognized for its various pharmacological activities and especially anti-snake venom property; however, the molecular basis for this property is not understood. It is reported that snake venom PLA2 is a toxic factor with pharmacological effects independent of their catalytic activity. Here we report the inhibition of catalytic property of Cobra and Viper (group I and group II) snake venom PLA2 by the phytocompounds from E. scaber through molecular docking and dynamics studies. Initially, Lipinski's rule, ADMET, and molecular docking studies were carried out. Our results show that among 124 phytocompounds, crepiside E (deacylcynaropicrin-3' beta-glucopyranoside) has shown interactions with the conserved catalytic active site residues, His 48 and Asp 49, in both the PLA2s. Further, molecular dynamic simulations for 60 ns confirmed the stability of crepiside E in the active site of PLA2s and were found to be stable throughout the simulation. In order to understand the drug-likeness of crepiside E, pIC50 and MMGBSA scores were correlated by performing a linear regression analysis. Crepiside E was found to have similar chemical features to that of doxycycline, a known PLA2 inhibitor as indicated by a similarity score of 64.15%. Hence, it is concluded that crepiside E beta glucopyranoside present in Elephantopus scaber contributes to neutralizing the snake venom.

Predictors of Poor Outcome in Neonates with Pyogenic Meningitis in a Level-Three Neonatal Intensive Care Unit of Developing Country.

BACKGROUND: The mortality of neonatal pyogenic meningitis is reduced to 10-15%, but morbidity is unchanged. METHODS: Primary objective is to determine the outcome, i.e. death or abnormal neurological examination (NE) at discharge and abnormal developmental quotient (DQ) at 3 months. Secondary objective is to find predictors of poor outcome. RESULTS: In all, 89 neonates enrolled, 10 expired and 24 neonates had abnormal NE at discharge. A total of 59 neonates came for follow up, 13 had DQ < 70. Prolonged shock (odds ratio, OR: 8.28; p = 0.001), coma (OR: 4.3; p = 0.001), seizures (OR: 14; p = 0.012), mechanical ventilation (OR: 18.55; p = 0.00), orogastric feeding (OR: 2.78; p = 0.042) and electroencephalography (EEG; OR: 9.6; p = 0.00) predicted poor short-term outcome. Abnormal NE at discharge (OR: 15.6; p = 0.001), EEG (OR = 10.60; p = 0.00) and brainstem-evoked reflex audiometry (OR = 37.20, p = 0.00) predicted a low DQ at 3 months. Mortality and morbidity of neonates with Pyogenic Meningitis (PM) were similar to that in developed countries. Outcome depended on severity of the disease and NE at discharge.

Chryseobacterium Indologenes-Associated Pneumonia in 2 Neonates.

Chryseobacterium species are widely distributed in nature and can rarely cause human infection. Few cases reported in hospitalized patients are in immunocompromised hosts with indwelling devices and associated comorbidities. Chryseobacterium species are usually multidrug resistant. We describe 2 cases of Chryseobacterium indologenes-associated pneumonia in neonates and review the published infant cases.

Feasibility study of an outreach program of newborn screening in Uttar Pradesh.

OBJECTIVES: To create awareness about newborn screening (NBS) in underprivileged population and establish the model of NBS as an outreach program in peripheral hospitals in and around Lucknow. METHODS: During this project, social workers were trained in taking informed consent, demographic details of newborns, heel prick samples and transporting the filter papers to a central laboratory. These social workers were posted in seven hospitals, in and around Lucknow, catering to families with low socioeconomic strata. Assays were performed in a single laboratory and results were conveyed back to the social workers and the hospitals. RESULTS: A total of 13,426 newborns were screened for the three conditions namely congenital hypothyroidism (CH), biotinidase deficiency and galactosemia. Over all coverage rate was 73.5 % and average time of reporting of results was 8.8 + 2.4 days. More than 85 % (86.7 %) families with positive screening test could be contacted back and out of them, 83.6 % babies could be sampled for confirmatory tests. Eleven babies were diagnosed to have CH. The study has shown positive and enthusiastic responses of the lay persons. Questionnaire based survey among lay persons showed that almost 100 % individuals understood the advantages and method of NBS. CONCLUSIONS: This project has been successful in establishing a model of NBS as an outreach program in and around a district.

Newborn screening for congenital hypothyroidism, galactosemia and biotinidase deficiency in Uttar Pradesh, India.

OBJECTIVE: To assess feasibility and recall rates for newborn screening for congenital hypothyroidism, galactosemia and biotinidase deficiency in a predominantly rural and inner city population in and around the City of Lucknow in Uttar Pradesh, India. DESIGN: Prospective observational study. SETTING: Two tertiary-care and 5 district hospitals in and around Lucknow. PARTICIPANTS: All babies born in above hospitals during the study period. METHODS: Heel prick samples were collected after 24 hours of life. Dried blood spot TSH, total galactose and biotinidase were assayed by immunofluorometry. Age related cut-offs were applied for recall for TSH. For galactosemia and biotinidase deficiency, manufacturer-suggested recall cut-offs used initially were modified after analysis of initial data. MAIN OUTCOME MEASURE: Recall rate for hypothyroidism, galactosemia and biotinidase deficiency. RESULTS: Screening was carried out for 13426 newborns, 73% of all deliveries. Eighty-five percent of those recalled for confirmatory sampling responded. Using fixed TSH cut off of 20 mIU/L yielded high recall rate of 1.39%, which decreased to 0.84% with use of age-related cut-offs. Mean TSH was higher in males, and in low birth weight and vaginally delivered babies. Eleven babies had congenital hypothyroidism. Recall rates with modified cut-offs for galactosemia and biotinidase deficiency were 0.32% and 0.16%, respectively. CONCLUSIONS: An outreach program for newborn screening can be successfully carried out in similar socio-cultural settings in India. For hypothyroidism, the high recall rate due to early discharge was addressed by age-related cut-offs.

Human parvovirus B19 associated dilated cardiomyopathy.

We describe two children presenting with acute left ventricular dysfunction. Both cases had evidence of dilated cardiomyopathy, requiring inotropic support and were tested for cardiotropic viruses by conventional or real-time polymerase chain reaction using specific primers for enteroviruses, human parvovirus B19, Adenoviruses, Epstein-Barr virus (EBV), herpes simplex viruses 1 and 2 (HSV), human herpes virus-6 (HHV-6) and cytomegalovirus (CMV). IgG and IgM antibodies against parvovirus B19, EBV, HSV and CMV were also tested by ELISA. One case tested positive for parvovirus B19 infection and recovered completely within 6 months in absence of any specific therapy. The other case tested positive for parvovirus B19 infection in association with hypocalcaemia and was cured following standard heart failure therapy along with calcium and vitamin D supplementation. Sequence analysis of DNA products from both patients revealed genotype 3. To best of our knowledge this is first report of circulating genotype 3 from India.

Respiratory distress including meconium aspiration syndrome in vigorous neonates born through meconium stained amniotic fluid: incidence, onset, severity and predictors at birth.

OBJECTIVE: This study aimed to find out incidence, predictors, onset and severity of respiratory distress including meconium aspiration syndrome (MAS) among vigorous neonates born through meconium stained amniotic fluid (MSAF), which may or may not be evident at birth. METHODS: Two hundred ninety vigorous neonates were studied. Data were collected on perinatal risk factors, clinical course and development of respiratory distress. Predictors of respiratory distress were identified by logistic regression and a score based on adjusted OR was assigned for each. Diagnostic performance of the score (0-24) was assessed on another 247 vigorous neonates using receiver operator characteristic analysis (ROC). RESULTS: Respiratory distress developed in 97(33.4 %) infants, MAS in 75(25.9 %). The distress appeared within 12 h in 97.9 %, was severe in only 21.7 %. Of 10 risk factors significantly associated with respiratory distress, seven entered in regression analysis. Fetal distress(adj OR = 11.8; 95%CI = 6.2-22.5), prolonged labor(adj OR = 5.2; 95%CI = 2.5-10.7), and absent/poor cry(adj OR = 5.6; 95%CI = 2.4-13.3) were identified as independent predictors; each assigned a score of 12, 6 and 6, respectively. To predict respiratory distress, a cut-off score of 9 points had sensitivity-74.1 % (95%CI = 63.3 %-82.7 %), specificity-84.6 % (95 % CI = 77.9 %-89.6 %), positive predictive value- 71.6 % (95%CI = 60.8 %-80.4 %), negative predictive value- 86.2 % (95 % CI = 79.6 %-90.9 %), likelihood ratio (LR) + ve 4.8(95%CI = 3.3-7.0) and LR-ve 0.3(95%CI = 0.2-0.4). CONCLUSIONS: Respiratory distress occurred in one third neonates, mostly had onset within 12 h of birth, and it was mild to moderate in majority. Fetal distress, prolonged labor, and absent/poor cry predicted respiratory distress and were validated. However, larger studies in different settings are required to confirm its utility.