Evaluation of Bacterial Population in the Saliva of Drug Addicts

Background: The human mouth harbors over 700 microbial genera. The large number of them is normal bacteria; few of them are opportunistic pathogens. Objective: The study was aimed to evaluate the bacterial population of drug addicted persons and compare it to that of a normal healthy person. Methods: Five different samples of human saliva; alcohol drunken person, oral moist snuff user, cigarette, hashish and shisha smoker were investigated the involvement of bacteria in them. One sample was collected from normal non addict person. The bacterial population in drug addictive persons were compared that on normal non addictive person. Results: The bacteria observed were: Gram negative cocci, mono cocci, streptococci, vibrio, gram negative rod, gram positive staphylococcus, gram positive streptococci and gram positive rod. Conclusion: The order of bacterial population was; alcohol drunken person<oral moist snuff user ˂ cigarette smoker ˂ hashish smoker ˂ shisha smoker˂ normal non addictive person. From the findings in this study it is concluded that all these drugs have some antibacterial activities. DOI : 10.14302/issn.2473-1005.jdoi-18-2115 Corresponding Author: Muhammad Zahoor, Department of Chemistry, University of Malakand ChakdaraDir (Lower), Pakistan, P.O Box: 18000. Email: mohammadzahoorus@yahoo.com


Background
Saliva is a watery fluid, which contains about 99 percent water.A large difference is present in the concentration of dissolved organic and inorganic solid in the saliva of two individuals as well as in the single individual.Human Saliva performs many biologic functions that are very important in maintaining oral health [1].
The human mouth harbors over 700 microbial genera [2,3].The large number of them is normal bacteria; few of them are opportunistic pathogens, which are important for the growth of mouth microbial contagious diseases for example as dental caries and periodontitis [4].These salivary bacteria reproduce the oral micro biota composition and can serve as a sign of the health and disease condition of oral cavity.For example, the bacteria present in the saliva counts have mostly been used for caries risk assessment [5][6][7].
There are nearly ten times as many bacterial cells in the human flora as there are human cells in the body.A number of them are present on the skin and as gut flora [8].The human mouth provides an ideal environment for the existence and growth of bacteria.It provides a source of water and nutrients, as well as an average temperature [9].Resident bacteria of the saliva adhere to teeth and gums to resist mechanical flushing from the mouth to stomach where they are destroyed by HCl [10].
The Saliva of a healthy man contains the bacteria like Streptococcus, Prevotella and Veillonella.Among them Streptococcus is the main species that accounted for 7 different species.Among the seven Streptococcus species, S. salivarius appeared as the most common species.Streptococcus and Prevotella are present in saliva of human from the ages of 32 to 35.
While the saliva of those human whose ages are from five to sixty five containing the genera Rothia.Saliva of young children and older human shows higher bacterial diversity than that of young adults [11].Anaerobic bacteria present in the oral cavity are Actinomyces, Arachnia, Bacteroides, Bifidobacterium, Eubacterium, Fusobacterium, Lactobacillus, Leptotrichia, Peptococcus, Peptostreptococcus, Propionibacterium, Selenomonas, Treponema, and Veillonella [11].
Streptococcus faecal is present in saliva is gram-positive cocci, while Eikenella and enterobacteriaceae are gram-negative bacilli, actinomyces and lactobacillus are gram-positive bacilli, veillonella is gram-negative cocci.
The aim of this study was evaluate the bacterial population of drug addicted persons and compare it to that of a normal healthy person.• The saliva of wine drunken.

All
• The saliva of oral moist snuff user.
• The saliva of hashish smoker.
• The saliva of cigarette smoker.
• The saliva of shisha smoker.
• The saliva of normal, non-addict person (control).
The saliva samples of each category volunteers were mixed and inoculated to sterilized nutrient agar Different colonies of bacteria were observed visually and were identified by using Gram staining.The bacterial populations in the selected samples were determined using hemocytometer.

Results
Bacterial strains present in selected mixed saliva samples were observed under microscope.The   From the findings in this study it is concluded that all these drugs have some antibacterial activities.However the already reported health problem associated with the use of these drugs should be kept in mind while using these substances.Also the bacteria present in the human saliva played an important role in human body and in the oral cavity and by using, drinking and smoking different types of addictive substances the types as well as the numbers and population of the bacteria are affected.Therefore it should be necessary and will be fruit full if these addictive substances are avoided and thus chemistry of saliva, especially the population of bacteria would not be affected.
experiments were carried out according to the Scientific Procedures Issue-1 of Animal Bylaws-2008 approved by the legal bodies of the University of Malakand Khyber Pakhtoonkhwa, Pakistan.The Ethical Committee of the Department of Biochemistry granted approval for conducting this study under the said protocols (Procedures Issue-1 of Animal Bylaws-2008).Approval of the collection procedure was taken from University Review Board for the protection of human research prior from starting collection of saliva samples from volunteers.The sample of human saliva used in this study were collected from volunteers using different addictive substances like wine, oral moist snuff, hashish, shisha and cigarette.Five samples were collected from normal non addict persons (control) also.A total of thirty different saliva samples were evaluated for the involvement of bacteria present in them.The samples were categorized into six groups.In each category there were five volunteers of nearly same age with difference of ±6 months.The samples were collected in sterilized glass vails and collection was made between 10 to 11 O'clock and inoculation to agar plates were made within two hours.The amount of alcohol etc in the saliva samples were not determined at the time of experiments.The details of which are given as follow: