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ISOLATION AND CHARACTERIZATION OF BACTERIA AND FUNGI ASSOCIATED WITH FRESH FROZEN FISH
ABSTRACT
Fish and fishery products have been documented as a major source of food borne pathogens of which bacteria and fungi play a major role. The fungal and bacterial organism contaminating apparently health and diseased (Scomber Scombrus) fish in Nasarawa, Nasarawa state was investigated. An invitro Assay was carried out to ascertain and identifies major bacterial and fungal contamination of frozen fish, which hitherto had constituted on important dieting intake of people of Nasarawa Nigeria. Fish samples collected were identified and the microbial load of the sample determined using spread plate method. Differentiation and characterization of various isolates were based on biochemical reactions and gram staining techniques. Frozen mackered fish (Scomber scombrus) was used for the study. The result revealed that staphylococcus aureus, Escherichia Coil, Salmonella Species and Shigella species are the bacteria associated with the contamination of the fish. Aspergillus, species Penicillin, species Rhizopus, species candida and mucor specie are the fungi associated with the contamination of the fish. The average number of colonies were 11, 21 and 45. The colony forming unit of O.5aiquote were 1.1 X i07, 2.1 x 106 and 4.5 x l0 isolation of fungi and Bacteria from Scomber Scombrus Meant for human consumption is alarming which can be source of fish borne infection. Attention should be paid to fish health in the community so as to conserve public health and safeguard the community health.
CHAPTER ONE
1.0 INTRODUCTION
Fish is one of the most important source of animal proteins available in the tropics and has been widely accepted as good source of protein and other elements for maintenance of healthy body (Al-jufalli and Opera, 2006). Fish is regarded as healthier meat due to its high content of long chain polysaturated fatty acids which are associated with improving health and preventing diseases of old age (yagoub & Ahmed, 2009).
The less developed countries capture 50% of the world harvest and a large proportion of the catch are consumed internally. In Nigeria, fish constitute 40% of the animal protein intake. Fish and fish products constitute an important part in the international trade, indicating increasing consumer interest in commodity. Generally, fish are good sources of vitamin B 12 and B6. It is also a good source of fluorine and iodine which are needed for the development of strong teeth and the prevention of goitre in man (youes & Bartram 2007). However, availability of these vital nutrients depends to a large extent on the methods of storage such as salting, roasting, drying, and freezing. Iced fish of different types are of great demands by the Nigerian consumers as a relatively cheaper source of animal protein. Fish and fish products are important, not only from the nutritional point of view but also as an item of international and foreign exchange carrier for a number of countries in the world (Yagoub, 2009).
Fish is a very perishable, high-protein food that typically contains a high level of free amino acids. Microbes rnetabolise these amino acids, producing ammonia, biogenic amines such as putrescine, histamine, cadaverine, organic acids, ketones and sulfur compounds (Emborg et al., 2005; Olatsdottir et al., 2005; Dalgard et al., 2006).
Microbiological quality is of importance to public health as it directly relates to spoilage of fish and becomes the cause of food poisoning. Microbial hazards causing infections and poor health are closely related to food safety concerned with animal proteins derived from marketed food- fish and fish products, meat and meat products. This creates a burning question for all consumers with a high risk commodity with regard to pathogenic bacteria & Fungi contaminations alarming to food safety challenge (Nilla et al., 2012). Refrigeration temperatures are also relevant because they are used by most households in Nigeria for temporary storage of fish (Oramadike et al., 201 0).Frozen state condition is also important since most fishes consumed in Nigeria are imported and usually come in frozen state (Oramadike et al., 2010).The quality of fish degrades due to a complete process in which physical, chemical and microbiological forms of deteriorations are implicated.
Therefore maintenance of quality is of utmost importance in the production and trade of fish and fish products. Although only a few infectious agents in fish are able to infect humans, some exceptions exists that may result in fatalities. However, the greatest risk to human health is due to the consumption of raw or insufficiently processed fish and fish products (Yagoub and Ahmed 2009).
According to the Centre For Food Safety And Applied Nutrition in Washington (2001),most fish related food borne illness are traced to Salmonella, Staphylococciisspp, Escherichia spp, Murcor spp penicillin spp, candida spp. Rhizopus and aspergillus. (Yagoub and Ahmed 2009). Spoilage bacteria differ somewhat for freshwater aid marine fish and for temperate and tropical water fish, storage and processing conditions also affect the predominant species on iced fish under aerobic conditions (Hozbor ci’ al., 2006). Packing under carbon dioxide and low concentration of sodium chloride favors the growth lactic acid bacteria and Photobacterium phosphoreuni. Microorganisms are found mostly on the skin, gills, operculum and intestines of life and newly caught fish. Fish contamination can be linked to raw material, personnel, processing tools such as forklifts through leakage, openings in buildings and pets. Some pathogens may even become established in the processing plants from niches where they can survive for a long period of time. The quality of our fish is of major concern to the food processors, consumers and public health authorities and provisions of safe, wholesome and acceptable fish and fish products to consumers and control of microorganisms is essential to meet these objectives.
The potentials of seafood to harbor microbial athogens and causing subsequent illness is well documented for both developed and developing countries (Younes and Bartram, 2007; Wright et cii., 2004). Water related diseases continue to be one of the major health problems globally (Adebayo et al., 2012). Okonko et al., (2009), reported that both bacteria and fungi are common flora of iced fish and fish related products during packaging. Majority of iced fish are sold in the open markets. Arannilewa et at., (2005) noted that protein decreased with increasing duration of frozen storage with fresh samples not frozen having higher protein content. Disadvantages such as product dehydration, rancidity, drip loss and product bleaching have an overall effect on the quality of frozen food. In spite of some disadvantages associated with frozen storage, freezing is .accepted as the effective way of preserving fish (Arannilewa e tal.,2005). The effect of physical conditions and chemical agents on the growth of microorganisms in fish has been investigated and well documented. In recent time, modern biotechnology have introduced new techniques that detect early fish contamination, improve taste, modify the quality of the fish, prolong the shelf life and also impact disease resistance to the fish Oramadike et a!., 2010).
Majority of the iced fish marketed in Nasarawa state are sold in open markets and so are exposed to dust and contaminated air, encouraging the growth of microorganisms which might facilitate increased transmission and prevalence of pathogens. Moreover poor personal hygiene, such as neglecting to wash hands after handling contaminated materials encourage contamination of iced fish with pathogenic organisms. This study is to assess the Microbial Load of fish sold at Nasarawa market in Nasarawa state. Also to isolate/identify the bacteria & fungi responsible for the contamination of frozen fish retailed at Nasarawa market
Bacteria
Bacteria are group of micro organism all of which lack a distinct nuclear membrane (hence are considered more primitive than animal and plant cells) and most of which have a cell wall of unique composition. Most bacteria are unicellular, the cells may be spherical (cocci) rod-shaped (bacillus) spiral (spirillum) comma-shaped (vibrio) or cork screw shaped (Geirocheate). Generally they range in size between 0.5 and 5nlm.
Fungi
Fungi can be single celled or very complex multicellular organisms. They are found in just about any habitat but most live on the land mainly in soil or plant material rather than in sea or fresh water. A very small number of fungi cause disease in animals
1.1 Statement of Problem
Fish is mostly spoiled by bacteria and fungi microbiological quality is of importance to public health as it directly relates to spoilage of fish and becomes the cause of food poisoning. Microbial hazard causing infection and poor health are closely related to food safety concerned with animal protein delivered from marketed fish and fish product meat and meat product. This creates a burning question for all consumers with a high risk commodity with regards to pathogenic bacteria continuous alarming to food safety challenge. The isolation and characterization of these spoilage organism will go a long way to profer solution no. mater how little to this problem.
1.2 Justification
The study will help in the acquisition of the knowledge of fungi and bacteria associated or involved with the spoilage of fish, factors that enhance spoilage of fish and the risk involved in the consumption of spoiled fish.
1.3 Scope of The Study
The project work is limited to the isolation and characterization of bacteria and fungi associated with fresh fish sold around Nasarawa Local Government. This will help in creasing the health awareness of the public on the damages of eating uncooked or under cooked fish. The project work would have covered the whole of Nasarawa State but it was not possible due to financial and time constraint
1.4 Aim of The Study
To isolate and identify micro-organism associated with contamination of frozen fresh fish
1.5 Objective of The Study
The objective of the study are:
i. To determine the microbial Load of the Fish
ii. To identify the type of bacteria and fungi associated with frozen fish commonly consumed by people in this locality
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