Isolation and diagnosis of bacteria and fungi from some areas of tikrit and some villages

2.1. The Aim of this Study

The purpose of this study was to evaluate the amount of fungi and bacteria that are causing air pollution in some locations of Tikrit, including the Al-Alam region, Al-Bu Ajil, Al-Buhyazaa, and the town of Al-Karaat [8].

3.1. Material and Methods

[9] The biology labs at Tikrit University in Iraq's College of Science conducted the current study. The purpose of the study was to evaluate the amount of bacteria and fungi that cause air pollution in a few different areas of Tikrit, including the Al-Alam area, Al-Bu Ajil, Al-Buhyazaa, and the village of Al-Karaat. Samples for the study were first collected in November and January of 2021–2022.

3.2. Macconkey Agar

To prepare this medium, dissolve 51.5 g of MacConkey agar in 1 liter of distilled water. Transfer the mixture into a sterilizing container. Distribute the mixture into petri dishes and store in the refrigerator at 4 oC until needed [1].

3.3. Nutrinet Agar

To prepare this media, dissolve 28 grams in one liter of distilled water, put it in a sterilizing container, pour it into sterile Petri dishes, and store it in the refrigerator until needed [1].

3.4. Eosin Methylene Blue

As directed by the manufacturer, 35.96 g of this medium are dissolved in one liter of distilled water to prepare it. It is then transferred into a sterilizing container, distributed among plates, and kept in the refrigerator until needed [10].

3.5. Potato Dextrose Agar

This medium is made by dissolving 39 grams of it in one liter of distilled water, then putting it in an antibiotic to sterilize it and an antibiotic such as an anti-tetracycline with the medium. After that, the dishes are distributed, the incubator is used to make sure there is no contamination, and the media is refrigerated until needed [1].

3.6. Sterilization

All culture media is sterilized by autoclave at a temperature of 121°C \ 15 bar for 15 minutes [11].

3.7. Sample Collection

In order to determine the kind and quantity of pollutants present in the atmosphere of the study areas, Nutrient Agar, Potato Agar, and Potato Dextrose Agar media were distributed throughout the villages of Al-Karaat, Tikrit, Al-Alam, Al-Bu Ajil, and Al-Buhyazaa. The petri dish was then closed and placed in the incubator after being left open for 15 minutes [12]. The number of colonies that grew on the aforementioned dishes was calculated by counting them and recording the growth [13].

3.8. Samples Culture

After the isolates from the study areas were gathered, they were cultivated on eosin methylene blue agar, MacConkeys agar, and incubated at 37 ºC for a full day. Following this, the first growth observations were recorded and used for diagnosis [14].

3.9. Cultural Characteristics

On Nutrient Agar, MacConkeys Agar, Eosin Methylene Blue, and Potato Dextrose Agar, the features of the developing colonies were observed, including their size, shape, color, and capacity for differential growth as well as their ability to ferment. Shape, color, size, ability to grow on one medium without needing another, and fermentation were among the attributes noted [15].

3.10. Microscopic Characteristics

The bacteria underwent gram stain staining, followed by a microscope examination to document their features, including size, type (positive or negative), and how their cells were arranged in a chain or doubled up [16]. Meanwhile, the fungi were dyed blue with lactophenol to observe their external structures, including thalus and clipboard spores, and how their cells were arranged in single or doubled up groups (Alhayfat).

4.1. Total Number of Colonies

4.2. Total Number of Fungi

Table 1 displays the findings of the current study. It shows that a variety of bacteria, including lactobacilli and klebsiella, grew most in November in Albu Ajil. This was attributed to the favorable environmental conditions, which included a temperature range of 12 to 43 C° and humidity fluctuations [17]. These species also increased in the spring and autumn because of the presence of virulent factors, such as toxins, capsules, and antibiotic resistance, which aid in their survival, spread, and infection [7].

The results showed that the months with the lowest growth ratio were December and November, when the temperature dropped due to the cold weather. Nonetheless, certain anaerobic bacteria were able to withstand inappropriate environmental conditions [8]. Because the formation of lactic acid lowers pH, which acts as an inhibiting substance, these bacteria are also important in the production of natural preservatives and foods [9].

However, the presence of farm animals in these villages is what caused the bacteria to spread throughout the Al-Bu Ajil and Al-Buhayaz area. They were spread through contaminated soil, milk products, and direct human contact [10]. Some of them, like Enterococcus, can grow in both rich and poor environments with oxygen because they can withstand high temperatures between 10 and 45 degrees Celsius, a pH range of 4.6 to 9.9, and high sodium chloride concentrations [3].

[11], [12] The results of the current study were displayed in Table 2, where it was found that the predominant fungus strain is related to the genera Penicillium and Rhizobus. Arthrographis is responsible for blue mold disease, which is the most common fruit disease that affects apples.

It also infects a variety of hosts, including pears, tomatoes, corn, and rice, with Albu Ajil and Al- Buhayaz having the highest ratio of growth due to the area's agricultural nature and humidity availability [13]. These fungi are abundant in agricultural areas because some of them can harm citrus fruits [14].

Because Rhizopus fungi are heterotrophic fungi, they feed on human and plant parasites and can be found in a variety of organic materials, including fruits and vegetables [8]. Arthrographis is a type of fungus that grows in wood, soil, marine sediments, and agricultural environments. Due to the abundance of animals and air pollution from dust and high humidity, it was the cause of infectious animal disease. The areas with the largest percentage of fungal growth were Albu Ajil and Al-Buhyazaa, and the areas with the lowest percentage were Tikrit, Alam, and Al-Karaat [18].