The measurement of the number of airborne microbes was conducted manually after the Petri plates were exposed to free air in 15 rooms at the Clinical Microbiology Laboratory of Dr. Soetomo Regional General Hospital Surabaya for 1 hour, with a distance of 1 meter from the wall and 1 meter from the floor. Then, the Petri plates for bacteria were incubated at 37°C for 1 day meanwhile the Petri plates for fungi were incubated for 5 days at room temperature.^14,15 The control Petri plates during the treatment was covered and incubated at the same temperature and time, where there was no growth of bacterial or fungal colonies found in all control Petri plates. This indicated that the growth media used was good and not contaminated.

On the calculation of airborne bacteria and fungi, it was found that the number of airborne microorganisms in the urine laboratory was 326 CFU/m³, the blood laboratory was 312 CFU/m³, the pus laboratory was 156 CFU/m³, the sputum laboratory was 234 CFU/m³, the tuberculosis laboratory was 182 CFU/m³, the Biomolecular laboratory was 1 78 CFU/m³, the biomolecular laboratory 2 was 104 CFU/m³, the laundry room was 13 CFU/m³, the identification room, the media room, as well as the meeting room each contained 78 CFU/m³, the reading room was 390 CFU/m³, the hall was 143 CFU/m³ and the Mycology laboratory was 273 CFU/m³. The number of airborne microorganisms for the areas or units in the laboratory is 200-500 CFU/m³, which is the maximum concentration of microorganisms per m3 of air according to the Regulation of the Minister of Health of the Republic of Indonesia No. 1204 /MENKES/SK/X/2004 concerning Hospital Environmental Health Requirements. Based on this reference, the number of airborne microorganisms in each room was still within safe threshold, yet it is likely that Laboratory Acquired Infections (LAI) or all infections acquired through the laboratory will occur. In Addition, the poor air quality in the hospital environment can cause nosocomial infections.¹⁶ One of the mediums in the environment where fungi, viruses and bacteria can live and grow is air. Since the presence of microorganisms in any area is considered indoor air pollution, the rooms with the number of microbes below the fixed threshold must be taken into account. The transmission of nosocomial diseases can occur through indoor air. The presence of germs in the air is one of the variables that determine air quality in the hospital. According to a research, 10-20% of nosocomial infections are caused by poor air quality. A research by Gilang in 2018 showed that air can be a channel for the spread of nosocomial infections.¹⁷ Even though there are many evidences showing that the environment can serve as a reservoir for various pathogenic agents, these environmental contaminants are the reason why infections are contracted in hospitals.¹⁸

The temperature in every room had been measured according to the reference of the Minister of Health of the Republic of Indonesia No. 1204 /MENKES/SK/X/2004 regarding Hospital Environmental Health Requirements, by using a calibrated tool. The measuring device was turned on and then left for 5 minutes until the temperature was stable, then the numbers listed were read.¹⁹ The temperature measured at the Clinical Microbiology Laboratory of Dr. Soetomo Regional General Hospital Surabaya was found in the range of 24.4-29.4°C, in the urine laboratory it was 28.3°C, in the blood laboratory the it was 26.5°C, in the pus laboratory the it was 28°C, in the sputum laboratory it was 28.1°C, in the Tuberculosis laboratory it was 29°C, in the biomolecular laboratory 1 it was 24.4°C, in the biomolecular laboratory 2 it was 29.6°C, in the laundry room it was 28.9°C, in the media room it was 26.7°C, in the identification room it 25.1°C, in the meeting room it was 25.7°C, in the reading room it was 26°C, in the admin room it was 26.3°C, in the hallway it was 26.5°C and in the Mycology laboratory it was 29.4°C, which when compared with the Regulation of the Minister of Health about Health Hospital Environment no. 7 of 2019 the optimum temperature allowed in the laboratory room is 20-22°C and the humidity in the laboratory is 40-60%,⁷ therefore the temperature in 15 rooms of the Clinical Microbiology laboratory at Dr. Soetomo Regional General Hospital Surabaya did not meet the optimal number.

Based on the analysis results of the correlation between temperature and the number of airborne bacterial and fungal colonies using simple correlation analysis, it can be concluded that there is no correlation between the independent variable, namely temperature, and the dependent variable, namely the number of airborne bacteria and fungi in the Clinical Microbiology Laboratory of Dr. Soetomo Regional General Hospital Surabaya. This can be caused by the air temperature in the Clinical Microbiology Laboratory room at Dr. Soetomo Regional General Hospital Surabaya ranged from 24.4-29.4°C, while the species/genus found was from the genus of Aspergillus species which can live optimally at temperatures of 35-37°C and are still able to grow at temperatures of 6-60°C. Each species/genus of fungi has different temperature levels to grow optimally in its environment.¹⁹

Airborne bacteria and fungi found in the rooms might not be caused by temperature and humidity values, but carried by visitors or room occupants.²⁰ It was proven in the reading room which had the highest number of airborne bacteria and fungi (352 CFU/m³). This room also had a lot of visitors every day compared to the other rooms which caused indoor air bacteria and fungi to stick to indoor furniture. The large number of furniture in the room made dust easy to stick and difficult to clean, therefore the airborne bacteria and fungi were easily carried along with the dust and allowed for an increasing number of airborne bacteria and fungi. This reading room consisted of things such as several books, bookshelves, chairs and cupboards, so that dust easily sticks to them and is difficult to clean. As a result, airborne bacteria and fungi were easily carried away with the dust and allowed for the growth of airborne bacteria and fungi.¹² As a comparison, the laundry room had the least number of airborne bacteria and fungi, namely 13 CFU/m³ where this room was rarely visited by laboratory personnel and did not have a lot of furniture.

The measurements in 15 rooms at the Clinical Microbiology Laboratory Dr. Soetomo Regional General Hospital Surabaya showed that there were three rooms that still met the maximum relative humidity standards, namely the Blood Laboratory with a relative humidity of 52%, Biomolecular 1 with a relative humidity of 53% and a Mycology laboratory with a relative humidity of 59%. Meanwhile, the other 12 rooms did not meet the optimum criteria for relative humidity, i.e. the relative humidity in the urine laboratory was 66%, the relative humidity in the pus laboratory was 63%, the relative humidity in the sputum laboratory was 73%, the relative humidity in the TB laboratory was 68%, the relative humidity in the biomolecular laboratory 2 was 65%, the laundry room obtained a relative humidity of 62%, the media room obtained a relative humidity of 66%, the identification room obtained a relative humidity of 65%, the meeting room obtained a relative humidity of 71%, the reading room obtained a relative humidity of 73%, the administration room obtained a relative humidity of 72% and the hallway obtained a relative humidity of 71%, where according to the Regulation of the Minister of Health regarding Hospital Environmental Health No. 7 in 2019, the humidity in the laboratory is 40-60%.⁷

This study showed that there is no correlation between relative humidity and the number of airborne bacteria and fungi, which is the same as the results of research by Sajaddi in 2016 and Hiwar in 2020, which also reported that there is no relationship between relative humidity and the number of microbes in the air.^10,11 Furthermore, research by Vindrahapsari in 2016 found that there is no relationship between the number of airborne bacteria and relative humidity.¹² Research by Gumiyarna in 2021 also found that in government rehabilitation therapy rooms, there is no correlation between the number of airborne microbes and room temperature or humidity.¹³ Research by Vindrahapsari Kamilia in 2016 regarding the factors related to the number of airborne microorganisms in private university classrooms in Jakarta which found a correlation between the physical quality of the rooms such as humidity and the presence of bacteria in the rooms is another study that does not support this research.²¹ Other research which was conducted by Syahrul in 2018 to figure out the correlation between the physical environment and the quantity of germs in Haji Makassar General Hospital rooms concluded that room humidity is directly related to the quantity of germs. In addition, Research conducted by Arief in 2019 stated that humidity and temperature support the growth of bacteria.^22,23

The range of temperature in the Surabaya Clinical Microbiology laboratory rooms which was reached (24.4-29.4°C) was very possible for the viability of the Staphylococcus aureus bacteria to be found, because these bacteria can grow at temperatures of 8-50°C and had an optimal temperature for growth at 37°C.²⁴ Staphylococcus aureus is the most common opportunistic pathogen in humans. In addition, this bacterium is associated with a wide variety of diseases, ranging from mild skin infections to serious conditions that can turn fatal. Folliculitis, furuncles, boils, impetigo, mastitis and wound infections can be caused by this bacterium. There are also a number of additional infections, including toxic shock syndrome, necrotizing fasciitis, sepsis, septic thrombophlebitis, bacteraemia, pneumonia, endocarditis, osteomyelitis, meningitis, urinary tract infection and septic thrombophlebitis.²⁵

In this study, Aspergillus species was also identified. Depending on the species, colonies of Aspergillus species can have different morphology. The majority of species begins as white colonies which turn immediately into green, yellow, orange, black or brown. Colonies develop from downy to velvety in 3-5 days. While some have even colors, some have color patterns that form concentric circles. For microscopic morphology, the hyphae are skeletal and branch at an angle of 45°C in the tissue samples. From the hyphae, long conidiophores arise from the foot cells and end in growing vesicles. Some environmental species produce hyphae that are spiral or zigzag in shape. All can produce vesicles, which can be club-shaped, round, or semi-circular. Phialides, which can be found individually or in pairs, joined end to end covering the vesicles. Conidia, which form chains, come from the phialides. The round conidia may have a smooth or bumpy surface.²⁶ Distal lateral subungual onychomycosis, proximal subungual onychomycosis, otomycosis and cutaneous aspergillosis are a group of superficial and cutaneous mycoses that can be caused by Aspergillus species. Most Aspergillus skin infections are nosocomial in nature, especially in infants and patients with compromised immune systems, as well as after medical procedures such as surgery, catheter insertion or after occlusive dressings on burn victims. However, trauma and skin damage are also significant risk factors for the development of superficial and cutaneous Aspergillosis symptoms.²⁷