Hospital-based sentinel surveillance for Streptococcus pneumoniae and other invasive bacterial diseases in India (HBSSPIBD): design and methodology.

INTRODUCTION: Streptococcus pneumoniae is one of the frequently isolated organisms and an important aetiological agent of invasive bacterial diseases (IBD) like pneumonia, meningitis and sepsis. As a measure to control the burden of IBD, the Government of India introduced Pneumoccocal Conjugate Vaccine-13 (PCV-13) in the Universal Immunization Program in high burden districts of five states in a phased manner from 2017 onwards. It is essential to understand the trend of circulating pneumococcal serotypes associated with IBD in the prevaccination and postvaccination scenarios to decide on the expansion of vaccination programmes and PCV reformulation. This manuscript describes the protocol for hospital-based sentinel surveillance for S. pneumoniae and other organisms causing IBD across various geographical regions in India. METHODS AND ANALYSIS: Hospital-based surveillance is established in selected hospitals to recruit children aged 1-59 months with symptoms of pneumonia and other IBD. Diagnostic criteria were adapted from standard WHO case definitions. Case Report Forms (CRFs) are used to collect data from the enrolled children. Blood, cerebrospinal fluid (CSF) and other normally sterile body fluids are collected and subjected to microscopy, cytology, latex agglutination, biochemistry, bacteriological culture and real-time PCR as applicable. Pneumococcal isolates are serotyped and tested for assessing antimicrobial resistance patterns. Data will be analysed by simple descriptive statistics to estimate the proportion of pneumonia and other IBD due to S. pneumoniae, Hemophilus influenzae type b and Neisseria meningitidis. Prevalence of bacterial infection, circulating pneumococcal serotypes, antibiotic resistance patterns, serotype variability across seasons and regions will be described in terms of percentage with 95% confidence interval. ETHICS AND DISSEMINATION: The institutional review boards of the coordinating centre, all sentinel sites, regional and national reference laboratories approved the project. The results will be published in peer-reviewed journals and shared with stakeholders for deciding on revising vaccination strategy appropriately.

Exploring the potential hazard of Mycobacterium avium subspecies paratuberculosis as a cause for Crohn's disease.

BACKGROUND: Mycobacterium avium subspecies paratuberculosis (MAP) is the causative agent of Johne's disease (JD), or paratuberculosis in ruminants has been suspected to be implicated in the pathogenesis of Crohn's disease (CD) in humans with chronic inflammatory intestinal changes. As the hypothesis is now fast being recognized that MAP could possibly be the etiological agent of CD which is found to be excreted in milk of dairy animals subclinically or terminally ill with JD. AIM: The present study was aimed to detect MAP in milk by polymerase chain reaction (PCR) targeting IS900 and to describe the excretion pattern of MAP in milk from asymptomatic lactating cows and does with relevance to the public health significance. MATERIALS AND METHODS: A total of 77 milk samples were collected randomly from lactating animals which include cows (45) and does (32). All the 77 milk samples were processed to identify the presence of MAP by employing the direct IS900 PCR as per the standard protocol. RESULTS: Out of 77 milk samples from asymptomatic lactating animals, 12 (15.58%) were showed positivity for IS900 PCR in which 5 (11.11%) were from lactating cows and 7 (21.87%) were from lactating does. CONCLUSION: In our study, 15.58% of milk samples showed IS900 positivity which indicates the presence of subclinical MAP infection in lactating animals. Hence, there is a possibility for excretion of MAP through milk which can be a potential threat for CD in humans by raw milk consumption. Therefore, the prevention of MAP in the food chain need to be assured by sourcing raw products from animal herds free of MAP infection.

Prevalence of antibiotic resistant Staphylococcus aureus from raw milk samples collected from the local vendors in the region of Tirupathi, India.

AIM: The study was carried out with the aim to identify the suitability of the milk for consumer use with special reference to Staphylococcus aureus from milk samples collected from various local vendors and determine the antibiotic susceptibility pattern of those positive isolates. MATERIALS AND METHODS: A total of 110 milk samples were collected from the local milk vendors in and around Tirupathi region of India. All the samples were enriched with buffered peptone water in 9:1 ratio and the then inoculated on baird parker agar medium with added 2% egg yolk tellurite emulsion as selective medium for S. aureus and confirmed with mannitol salt agar, Gram's staining and biochemical tests. The typical cultural characters with coagulase-positive samples were taken as positive samples the positive samples were tested for antibiotic susceptibility with 10 different antibiotics by employing disc diffusion method. RESULTS: Prevalence of coagulase-positive S. aureus was 39.09% (43/110) from the milk samples. The antibiotic susceptibility test of positive isolates showed high resistant toward penicillin G 37/43 (86.04%) and ampicillin 32/43 (74.42%), and also showed resistant to methicillin 6/43 (13.95%), cephalothin 6/43 (13.95%), tetracycline 6/43 (13.95%), ciprofloxacin 4/43 (9.30%), enrofloxacin 3/43 (6.97%), cefoxitin 2/43 (4.65%), gentamicin 2/43 (4.65%), and co-trimoxazole 2/43 (4.65%). Many individual isolates showed resistant against two or more antibiotics in our study. CONCLUSION: The above study results show that the milk samples collected from local vendor having S. aureus, which can induce disease condition as well as antibiotic resistant to the humans particularly young children and old age peoples by means of consumption of raw milk and its products. This is the public health issue, which needs to be solved by educating the local vendors regarding health problems related to unhygienic milk supply and make the awareness among the consumers about this hazards and preventive measures.