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Extramural Aero-bacteriological Quality of Hospital Environment

Extramural Aero-bacteriological Quality of Hospital Environment

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Published by Aditya Pathak

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Published by: Aditya Pathak on Dec 14, 2011
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 Society of Applied Sciences
Extramural Aero-bacteriological Quality of HospitalEnvironment
Apurva K.Pathak* and Karuna S. Verma
*Deptt. of Pathology & Microbiology, Modern Dental College & Research Center, Indore(M.P.)-453112, India, e-mail: pathak.apurva@gmail.com.
Aeroallergens and Immunology LaboratoryDepartment of Biological Sciences, Rani Durgavati University, Jabalpur (M.P.) India -482001
A hospital produces waste by giving their service to the patients. This waste can be produced directlyin combination with the service (e.g. injection) or in the upstream (e.g. blood or urine cultures in thelaboratory) or downstream (vaccine) process. One kind of typical diseases treated in hospitals isinfectious diseases. By the services for known or unknown infectious patients, waste can be thesource of an infectious agent. If the microorganisms have survived the aerial transport, they can growand reproduce in the new environment and may spread further. If a living organism inhales theairborne particles, their fate will depend on many factors. Large particles, i.e. larger than 5 mdiameters, easily adhere to the mucus membranes of the upper respiratory tracts. Continuouslymoving cilia guide the particles to the throat, where they removed by coughing or swallowing.Particles smaller than 5 mm inhaled into the deeper parts of the lungs, many bacteria and viruses arethus well adapted by their size to reach the alveoli [1]. The lungs represent one of the largestinterfaces between the human organism and its environment, and thus a major site of interaction withmicroorganisms and its products. Infection of the lung is one of the most frequent causes of morbidityand death. Toxic substances can enter into the lungs either during infection by Gram-negative bacteria(40–60% of all cultured-diagnosed pneumonia), or when carried by inhaled airborne particles.Hospital environment consisted high level of potentially hazardous bacteria, fungi and other allergenic and / or immuno-toxic agents. These agents may penetrates into lungs of exposedresidences or dwellers and evoke inflammatory reaction leading to respiratory disease, such asasthma, mucous membrane irritation, allergic alveolitis etc. Hospital waste is not necessary a sourceof infectious agents. Only if the waste suspected to contain infectious agents like bacteria viruses, parasites, or fungi in a sufficient concentration it will be infectious waste. Members of the
groups, and
 Pseudomonas aeruginosa
, flavobacteria, andother non-fermentative bacilli can able to grow well in environmental fluids and some entericorganisms (e.g.,
 Escherichia coli
and Proteus spp.) that have limited powers of multiplication atenvironmental sites, are the potential source of infection, once these bacteria are introduced into the
 ASIAN J. EXP. BIOL. SCI. VOl 1 (1) 2010:128-135 ABSTRACT 
 An extramural aero-bacteriological study was undertaken to determine typical concentrations of airborneculturable bacteria at the vicinity of a local hospital in the tropical environment of India. Hospital acquired infection is a common phenomenon; however, hospital as a source of infection for the surroundings community is yet to be established. Present aero-bacteriological investigation includesenumeration, identification, and numerical analysis of air borne culturable bacteria in hospital associated environment. Bacteria isolated from the environment during the present study wererepresentative of normal micro flora of the skin, respiratory and gastro-intestinal tracts; it also includesthe opportunistic pathogens like Acinetobacter and Flavobacterium species with the environmental non- pathogenic genera. In this study respirable and nonrespirable fraction of bacteria were also quantify.The Pearson correlation study shows that both temperature and humidity have an effect on tenacity and  prevalence of the allbacterial type. A regression model with 67.15 % variance is also prepared in order to predict the bio-load for this atmosphere.
Aerosolization, Aero-bacteriological investigation, Enterobacteriaceae and Prediction.
environments from the patients. As a result, large populations built up, and may subsequently survivefor a very long time. All the Microorganism of ambient environment must not necessarily behazardous for humans; they are even a part of the natural micro flora. Changing environmental microflora can change the human normal micro flora too, as the human being is a part of its larger ecosystem. The immune system ( Innate and acquired) has an ability to maintain the healthy state of the organism, but this system under certain state ( e.g. Immunocompromised, old age, contaminationetc.) can be exposed of a massive attack of pathogen agents, protective barriers can break or are broken and the natural balance can be shifted [2].The contribution of airborne microorganisms to thespread of infection is likely to be greater than is currently recognized. This is partly because manyairborne microorganisms remain viable while being non-culturable, with the result that they are notdetected, and because some infections arising from contact transmission involve the airbornetransportation of microorganisms onto inanimate surfaces. [3] The aerosolized spore forming gram positive bacteria are able to survived in air for a long duration and gram negative non-spore forming bacteria can also able to survive in air up to 390 minute as a half life time recorded previously by theworkers [4]. Things are gone worse when these microorganism able to multiply in these aerosols [5].The transportation and forecasting of air borne microorganism is of prime importance toepidemiologist in order to access the chances of outbreak of disease in community environment and toevaluate the degree of pathogenicity of these organism. This comprehensive study has been made toevaluate the quantity, quality, respirable and non-respirable fraction of potentially pathogenic bacteriafor this environment. The effects of environmental factors on the total airborne bacterial bio-load alsoanalyzed by using correlation analysis and a regression model are prepared for this environment.Many works previously been done to enumerate and identify the air borne bacteria of nosocomialenvironment [6-11]. On the contrary, very few of them essentially concerned their study as thehospital as a source of bio-pollutants for the community [12, 13]. This present study deals with thetypes of bacterial flora originated from hospital or hospital associated waste, which disseminatesextramurally posing both occupational, and community related health problem.
Jabalpur (Latitude: 23.2; Longitude: 79.95; Altitude: 391.) is the third biggest city of MadhyaPradesh. The city of Jabalpur is set in a most attractive stretch of country. The metropolis itself standson a rocky stretch of land about 9.6 km. from the River Narmada and 20.8 km. from the marble rocksof 
. Jabalpur is one of the central districts of India. The city consists of long narrow plainsrunning northeast and southwest, and shut in all sides by highlands farming an offshoot from the greatvalley of the River Narmada. The hilly tracts in and around Jabalpur are covered by luxuriantvegetations. The climate of Jabalpur is overall pleasant and salubrious, has a year-round tropicalclimate generally characterized by warm days and cooler evenings.
Sampling Site
Medical college hospital is 750 bedded; one of the biggest tertiary care hospital of 
Region. The Medical College surrounded by typical urban mixed type habitation. A large number of  people from different region visited here not only for the indoor treatment but also as outdoor  patients. The kith and kin of these patients were used to stay outside within the premises of thehospital were also responsible for generating waste of various types. The aero-bacteriologicalsampling had been done within the premises of medical college hospital, 50 -100 meter apart from the building and at the Devtal area which is situated two kilometer from the hospital area as a control, induplicate and fortnightly in order to cover all the major season. The metrological data collected fromweather station Jabalpur. Apart from temperature and humidity five other metrological parameter (Mean sea level pressure (mb); Precipitation amount (mm), Mean wind speed (Km/h); Maximumsustained wind speed (Km/h); Maximum wind gust (Km/h); Indicator for occurrence of: Rain or Drizzle) were also recorded in order to analyze their effect on airborne bacterial load.
Isolation from Air
A bioaerosol may need to monitored, not only for mass or number concentration, but also for viability. The amount or proportion of viable microorganisms in the atmosphere can be the criticalfactor of interest. Thus, the sampling method selected should impose minimum stress on themicroorganism and supply nutrients to preserve viability. [14]. Area sampling should always becarried out near the potential sources of bioaerosols such as air supply systems, machinery and at or 
Extramural Aero-bacteriological Quality of Hospital Environment Apurva K.Pathak & Karuna S. Verma
near the workers’ position; Duplicate samples in time and place must always be taken [15]. TheAndersen 2-stage viable (microbial) particle sampler (2-STG) used for sampling which has beendeveloped for monitoring bioaerosols. It is a multi-orifice, cascade impactor with 400 holes per stage,drawing air at a flow rate of 28.3 L min-1. This sampling rate is comparative to the breathing rate of a person going about their normal work. However, worker respiration rates will rise with increasedmetabolic rate. In women, respiration rates are lower [16]. The different stages separate the airborne particles in size fractions. The stages have 50% cut-off diameters of 0.6 to 7 μm (depending on theorifice used) and the impaction holes arranged in a regular pattern that facilitates counting of colonies[17]. As the air velocity increases across the different impacting surfaces, the smaller particlesdeposited resulting in the upper stages collecting the larger particles while the lower stages collect thesmaller particles [18].The organisms deposited because the air forced to make a 90 degree turn to passaround the edge of the plate. This allows the smaller particles to pass around the growth mediumwhile the larger particles cannot make the turn and impact on the surface [16]. Dividing bioaerosolsinto size categories is important for epidemiological research [19]. Culture methods were usedeverywhere for the measurements of airborne microorganisms in the work environment [20]. For thisstudy, air sampling done on Tryptone Glucose Yeast Extract (TGYE) Agar Medium (Hi Media), andEosin Methylene Blue (EMB) Agar Medium (Hi Media) with the help of modified two stagesAndersen Sampler [21, 22]. The sampler placed at one meter height from the ground; operated for twominutes at both the site in duplicate at morning hour before the OPD started. For enumeration andidentification of total viable type of bacterial population present in air, the TGYE medium plate werekept on upper stage of the sampler, whereas for enumeration and isolation of respirable fraction of gram- negative bacteria, the EMB media plate were kept on lower stage of the sampler.
Isolation from Sources
Effective source control is the key issue in all matters concerning hygiene and hospitals are by nomeans an exception, [23] for this the source must be identified. This sampling was carry out toidentify the presence of the bacteria in dust, decayed materials, and debris of hospital residuesoriginated by the community and compare with those present in air. Water samples collected fromassociated sewage system of hospital, by holding the glass stopper, sterile bottle near its base in thehand and plugging it (necked downward below the surface) and transported to the laboratory in anicebox to avoid unpredictable changes in physiochemical as well as bacteriological characteristics.The dust and top soil of debris sampled in sterile polythene airtight bags. Processing of samples wasdone by serial dilution technique (10-2 to 10-4) to get only a few cells per ml. One ml of inoculumsfrom each dilution poured onto sterilized Petri plates of respective media (TGYE & EMB) at 45 0C by using Pour plate technique [24] and incubated at 37 ± 2 0C for 24 to 48 hrs.
Air Contamination Standards
The level of bacterial contamination of air is usually expressed in terms of number of bacteria-carrying particles per m3 (bcp m-3) or the bioload (B). B can be calculated from the below formula:
mRT1000N B
Where N is the number of colonies counted on the sample plate, T is the duration of the test in min,and R is the air-sampling rate in liters/min. (25). The recommended threshold value (TLV) for bioloadis 50 CFU/m3 (WHO).
Identification of Isolates
Bacteria can be identified by morphology, gram-staining, growth on specific substances and under special conditions, and production of specific metabolites [17]. After gram staining of bacteria, astudy by Krahmer et al. [26] divided the results into four categories as actinomycetes, gram-positiverods, gram-negative rods, and gram positive cocci. In this study, identification of isolates was done byusing standard methods and manuals[25, 27-31].
Statistical Analysis
The number of samples collected will influence the precision of the exposure estimate and theassociated confidence limits [32]. Monitoring environments can be difficult and time consuming,which can lead to a small number of samples being collected [33]. In order to analyze the effect of various environmental factors on the prevalence of airborne bacterial population and degree of its
Extramural Aero-bacteriological Quality of Hospital Environment Apurva K.Pathak & Karuna S. Verma

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