EFFECT OF DEPURATION AND SOAKING ON THE HEAVY METALS, NUTRIENT COMPOSITION AND MICROBIAL LOAD OF SHELLFISH HARVESTED FROM POLLUTED CREEK OF RIVERS STATE, NIGERIA

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ABSTRACT


Water, sediment and shellfish were collected from four locations (Slaughter, Aboluma, Okwujiagu and Oginiba) of the Azobie creek of River state Nigeria and were evaluated for microbiological, heavy metals, nutritional composition and physico-chemical parameters. Bacterial isolates consisting of  ( total  heterotrophic bacteria and hydrocarbon utilizing bacteria count)  and fungal count (total fungal count and hydrocarbon utilizing fungi count) were analysed from the shellfish and  its habitant (water and sediments). Data generated were subjected to analysis of variance (ANOVA) and mean were separated using fishers minitab 2017. The highest total coliform counts obtained from  Slaughter (control) had a value of 1.52 during wet season and 1.23x105 CFU/g in dry season and in Aboloma it was 1.53x105CFU/g for wet season and 1.12x10Cfu/g for dry season in periwinkle . These  decreased to an acceptable limit of 0.02 x10 5CFU/g at different time intervals of 24-96h of soaking and depuration. Total heterotrophic bacteria count in periwinkle had the highest count in control at Slaughter, Oginigba and Abuloma at a value of 0.48 and 0.33X105 CFU/g This was reduced to acceptable limit of 0.12 x 105 CFU/g. Similar trend occurred in oyster sample at the same locations. Total hydrocarbon utilizing bacteria and fungal count in oyster had the highest count of 9.80 during wet season and 8.90x10CFU/g for dry seasons at Slaughter and Abuloma at 9.17x10 CFU/g for wet season and 7.12x10CFU/g during dry season. Oginigba did not record any bacteria count. Total faecal  count was reduced at different time interval of 24-96h to an acceptable limit , while for depurated at different time intervals during dry season, it was not reduced. This could be as a result of the different depuration methods. The same trend was observed in periwinkle although it was not fast as in oyster. The different bacterial profiles are probably attributable to industrial activities of the sites. The moisture, protein, fat, crude fiber and ash content were the major constituents which had been considered in evaluating the nutritional value of these species. Soaking  shellfish had no effect on the proximate composition except in fat where it increased  the fat and protein content in the control. Physio chemical parameters varied among different locations. The levels of Cd ( 0.00-2.5ppm) recorded in these samples were higher than the maximum recommended limit of 2-0ppm in sea food. The value of Cr recorded was between 1.50 – 2.0 ppm and was found to be higher than the maximum recommended limits of 0.15-1.0ppm.The levels of Pb (0.001-2.50) recorded in this study was higher than the recommended limits of 2.0ppm  in some locations. The micro nutrient (K, Fe, and Zn) were above the permissible limits for human consumption as recorded by the Food and Agricultural Organization. Depuration had little effect on  contaminant while soaking had no effect at different locations and different time intervals. Depuration performed better than soaking during dry season than raining seasons. Oyster was faster than periwinkle in removing this contaminant during depuration than soaking in during dry seasons.







TABLE OF CONTENTS

                                                                                                                               

Title page                                                                                                                                                            ii

Declaration page                                                                                                                                              iii

Certification                                                                                                                                                       iv

Dedication                                                                                                                                                          v

Acknowledgements                                                                                                                                       vi

Table of contents                                                                                                                                             vii

List of Tables                                                                                                                                                      xiv

List of    figures                                                                                                                                 xiv

List of plates                                                                                                                                                       xxi

Abstract                                                                                                                                                               xxii                                                                                                                                                        

CHAPTER 1: INTRODUCTION                                                                                                      

1.1 Background of the study                                                                                                                       1

1.2          Statement of Problem                                                                                                                   5

1.3          Justification of the Study                                                                                                              6

1.3          Objectives of the Study                                                                                                7

1.3.1 General objective of the study                                                                                                       7

1.3.2 The specific objectives of the study                                                                                              7

CHAPTER 2

2.0       Review of Related literature                                                                                                          9

2.1       Sea Food:  Meaning and Component                                                                                         9

2.2          Sources, Distribution and Components of Shellfish                                                           9

2.3          Shellfish Nutritional Value and Food Uses                                                                             10

2.4          Food Uses and Industrial Application of Shell Fish                                                             15

2.5          Creeks of River State- Geographical location, Vegetation and Weather

Condition                                                                                                                                            15

2.6          Industries in the Creeks-Distribution, Raw Materials, Human Faeces        17

Discharge and Management in creek                                                                                   16 

2.7          Impact of Waste Discharge into the Water Bodies on Aquatic and Human             

life in the Creek                                                                                                                                19

2.8          Microbial Contaminants in the Water and their Impact on Human Health               19

2.9          Shellfish and Heavy Metal Absorption                                                                    22

2.10        Nutritional Composition of Seafood                                                                                        23

2.11        Reduction of risks Associated With Contaminated Shellfish                          24

2.12        Shellfish Contamination                                                                                                                24

2.13        Infectious Risk Related to Shell Fish Consumption                                                             25

2.14        Pathogenic Organisms Associated In Shellfish Disease Outbreaks                              27

2.15        Disease outbreaks associated with shellfish                                                                     31

2.16        Proximate Composition Of Shellfish                                                                                        31

2.17        Depuration and Relaying (Soaking)                                                                                          32          

2.18     Origin of Depuration                                                                                                                         34

2.19     Pollution limit for Shellfish Depuration and Relaying                                           36                      

2.20       Basic Principles of Depuration                                                                                     36

2.21       Quality of a Good Depuration Stand                                                                         37          

2.22         Heavy Metals                                                                                                                   37

2.23        Cadmium Toxicity                                                                                                                            42

2.24           Effects of Heavy Metals                                                                                                             43

 

2.25        Types Of Heavy Metals That Can Be Dangerous To The Body                       44

           

CHAPTER THREE

3.0          Materials and Methods                                                                                                 48

3.1          Shellfish                                                                                                                                               48

3.2          Study Area Description                                                                                                  48           

3.3          Water Sample Collection                                                                                                              49

3.4          Collection of Sediments                                                                                                                50

3.5          Shellfish Collection                                                                                                                          51

3.6          Stocking and Managing the Shellfish in the Soaking and Depuration Tanks             52

3.6          Determination of Heavy Metals in Soft tissue of Shellfish                                              57

3.7          Determination of Heavy Metals in Sediments                                                     57

3.8          Determination of Heavy Metals in Water                                                                              58

3.9          Analysis of Physical and Chemical Parameters of the Sampling Area Water

sample                                                                                                                                 59

3.9.1      Temperature                                                                                                                                     59

3.9.2      pH                                                                                                                                                          59

3.9.3      Salinity                                                                                                                                  60

3.9.4      Turbidity                                                                                                                                              60

3.9.5      Dissolved oxygen (DO)                                                                                                  60

3.10        Proximate Analysis                                                                                                                          62

3.10.1    Moisture content analysis:                                                                              62

3.10.2    Ash content analysis:                                                                                      62


3.10.3Fat content analysis:                                                                                       63

3.10.4    Crude protein analysis:                                                                                                  63

3.10.5    Carbohydrate analysis                                                                                                               64

3.10.6    Crude fibre analysis                                                                                                                        64

3.11        Microbiological Analysis                                                                                                                65

3.11.1    Sample preparation                                                                                                                        65

3.11.2    Total heterotrophic bacteria count                                                                                           65

3.11.3    Hydrocarbon utilizing bacteria count                                                                                    66

3.11.4    Total heterotrophic fungal count                                                                                              66

3.11.5    Hydrocarbon utilizing fungi count                                                                                             66

3.11.6    Vibrio count                                                                                                                                       67

3.11.7    Faecal coliform count                                                                                                                 67

3.11.8    Isolation and purification                                                                                                              67

3.11.9    Identification of microorganisms                                                                                               67

3.12        Identification and characterization of the isolates                                                             69

3.12.1    Bio chemical test                                                                                                                              69

3.12.2    Catalase Test                                                                                                                                     70

3.12.3    Coagulase test                                                                                                                                  70

3.12.3    Motility in dole urease (MIU) agar.                                                                                          70

3.12.4    Preparation of MIU agar                                                                                                               71

3.12.5    Klinger Iron Agar (KIA)                                                                                                   71

3.12.6    Preparation of KIA                                                                                                                           72

3.12.7    Inoculation                                                                                                                                         72

3.12.8    Preparation of simmons citrate                                                                                 72

3.12.9    Oxidase                                                                                                                                                72

3.12.10 Indole                                                                                                                                                    73

3.12.11 Methyl red                                                                                                                                          73

3.12.12 Voges Proskauer                                                                                                                              73

3.12.13 Urea utilization                                                                                                                  74

3.12. 14 Citrate                                                                                                                                  74

3.13        Molecular Analysis                                                                                                                          75

CHAPTER 4: RESULT AND DISCUSSION

4.1          Effect of Soaking and Depuration Time on Bacterial load of Oyster

Harvested From Different locations of the Creek at Different Seasons of

The Year                                                                                                                                              80

4.2          Effect of Different Time Intervals of Soaking and Depuration at

Different Locations on Total Heterotrophic Bacteria (THBC) X105cfu/G

Of Periwinkle Sample at Different Season                                                                            83

4.3          Effects of Different Time Intervals of Soaking and Depuration on

Hydrocarbon Utilizing Bacteria (HUBC) x105Cfu/g at Different location in

Oyster Sample at Different season x105Cfu/g                                                     86

4.4          Effects Of Different of Soaking and Depuration Time on Total Fungal

counts (Tfc) X105cfu/G of Periwinkle Sample Harvested from Different

location at Different Season                                                                                                     89

4.5          Effect of different Time Interval of Soaking And Depuration on Total

Fungal counts (Tfc) X105cfu/G Of Oyster Sample Harvested from

Different location at Different Season                                                                    93            

4.6          Effect of Different Time Interval of Soaking and Depuration Methods at

Different locations on Hydrocarbon Utilizing Fcounts (HUFC)

X105cfu/G Of Periwinkle at Different Season                                                       97

4.7          Effects of Different Time Interval of Soaking and Depuration at Different

locations on Hydrocarbon Utilizing Fungal Counts (HUFC) x105cfu/g of

Oyster sample at Different Season x105CFU/g                                                    100           

4.8          Effect of Different Time Interval of Soaking and Depuration in Faecal

Coliform Count of Oyster Sample Harvested at Different Locations at Different

Season x105cfu/g                                                                                                                             104

4.9          Effect of Different Time Interval of Soaking and Depuration Methods in

Faecal Coliform Count of Oyster Sample Harvested from Different Locations at

Different Season x105cfu/g                                                                                                         107

4.9.1      Total load of different microbial contaminants on sediment and water Body

from different locations                                                                                                                110

4.10        Organisms Isolated                                                                                                                         122

4.11        Result and Discussion of Proximate Composition of Shellfish                      

Harvested at Different Location During Different Season                                               131

4.12        Physicochemical Parameter Of Water                                                                     150

4.13        Heavy Metal Concentration of Shellfish Harvested from Different

Locations at Different Seasons                                                                                   166

4.13.1    Effect of different time intervals of soaking and depuration on six heavy               166

metals for periwinkle at Abuloma location                                                                           

4.13.2    Effect of different time intervals of soaking and depuration on six heavy              

metals for periwinkle at slaughter location                                                                           170

4.13.3    Effects of different time intervals of soaking and depuration on six         

heavy metals for Periwinkles at Oginigba location                                                             172

4.13.4    Effect of different time intervals of soaking and depuration on six (6) heavy

Metal for Okwujiagu location                                                                                                     175

4.13.5    Effect of different time intervals of soaking and depuration on the level of

six heavy metals in oysters at different season for Slaughter location      180

4.13.6    The effect of different time   intervals of  soaking  and depuration on

six heavy metals of oyster at different season Aboluma location.                  183

4.13.7    The effects of different time intervals of soaking and depuration on six

heavy metal on oyster at different seasons for Okwujiagu.                                          187

4.13.8    Effects of different time intervals of soaking and depuration of oysters on

six heavy metal at Oginigba location different seasons                                   190

4.13.9 Heavy metal concentration in water and sediment                                                              196

 

CHAPTER 5: CONCLUSION AND RECOMMENDATIONS

5.1               Conclusion                                                                                                                                          198

5.2               Recommendation                                                                                                                            199

                References                                                                                                                                         201            

                Appendices                                                                                                                                        222      






  


LIST OF TABLES

                                                                                                                                                                                 

2.1          Vitamin contents of some shellfish meat                                                                              13

2.3          Mineral contents of some shellfish meat                                                                              14

2.4          Potential enteric and water-based microbial pathogens                                 20

2.5          Seafood borne illnesses associated with bacterial pathogens                                  21

2.6          Proximate composition of some shellfish species from Indian waters      33

4.1          Effect of different time interval of soaking and depuration at different

locations on total heterotrophic bacterial count (THBC) x105 CFU/g in

oyster sample at different Seasons                                                                                         82

4.2          Effect of different time intervals of soaking and depuration At different

locations on total heterotrophic bacteria (THBC) x105cfu/g

in periwinkle                                                                                                                                      85

4.3          Effects of different time intervals of soaking and depuration on

hydrocarbon utilizing bacteria (HUBC) x105Cfu/g at different location in

oyster sample at different season x105cfu/g                                                                  88

4.4          Effects of different time interval of soaking and depuration on total fungal

counts (TFC) x105cfu/g at different location in periwinkle sample at

different Season                                                                                                                              92

4.5          Effect of different time interval of soaking and depuration at different

locations on total fungal counts (TFC) x105cfu/g of oyster sample at

different Season                                                                                                                              96

4.6          Effect of different time interval of soaking and depuration methods at

different locations on hydrocarbon utilizing fungal counts (HUFC) x

105cfu/g in periwinkle at different Season                                                                            103

4.7          Effects of different time interval of soaking and depuration at different

locations on hydrocarbon utilizing fungal counts (HUFC) x105cfu/g in

oyster sample at different season x105cfu/g                                                                     106

4.8          Effect of different time interval of soaking and depuration methods in

faecal coliform count of oyster sample harvested from different location at

Different Season x105cfu/g                                                                                                             109

4.9          Effect of Different Time Interval of Soaking and Depuration Methods at

Different Locations on Faecal Coliform in Periwinkle Sample at Different

Season x105cfu/g                                                                                                                            

4.10        Biometric data of bacteria isolates                                                                                            128

4.11        Effect of different time intervals of soaking and depuration On Six          

Heavy metals for periwinkle at Abuloma Location                                                             133

4.12        Effect of different time intervals of soaking and depuration on six heavy

metals for periwinkle at slaughter location                                                                           134

4.13        Effect of different time intervals of soaking and depuration On six

heavy metals for periwinkle at Oginigba Location                                                              136

4.14        Effect of different time intervals of soaking and depuration of periwinkle

on Six (6) heavy metal for Okwujiagu Location                                                    137

4.15        Effect of different time intervals of soaking and depuration on the level of

six (6) heavy metals in myster at different season for slaughter location 139

4.16        Effect of different time intervals of soaking and depuration on six heavy

metals at different season Aboluma Location                                                                140                                                             

4.17        Effect of different time intervals of soaking and depuration on six heavy

metal on oyster at different seasons for Okwujiagu.                                                       142 

4.18        Effect of different time intervals of soaking and depuration on six heavy

 metal at Oginigba location different Seasons.                                                         143 

4.19        Heavy metal analysis in water and sediments                                                           145








LIST OF FIGURES


3.1          Map of the sampling stations along the Azonie Creek in River State          49

4.1          Total hetrotrophic bacteria count in sediment and water body from

different locations at different seasons                                                                 111        

4.2          Hydrocarbon utilizing bacteria in sediment and water body from different

locations at different seasons                                                                                                 113

4.3          Total fungal counts in sediment and water from different locations at

different seasons.                                                                                                                           115

4.4          HUF counts in sediment and water from different location at different 117

4.5          Total vibro parahaemolyticus  counts in sediment and water  from

different locations at different seasons                                                                 119        

4.6          Total Faecal coliform counts in sediment and water from different

locations at different seasons.                                                                                                   121

4.7          Escherichia Coli Strain                                                                                                    124

4.8          Vibrio Stylophorae Strain                                                                                                             125

4.9          Candida albican strain                                                                                                   125

4.10        Meiothermus grandicius Strain                                                                                  127

4.11        Biometric data of bacteria count.

                                                                                                                                                  130


4.11    The moisture content of soaked and depurated shellfish for different

                   locations at different time intervals for raining  seasons                              133

4.12        The moisture content of soaked and depurated shellfish for different

locations at different time intervals for dry seasons                                                              134

4.13        The fat content of soaked and depurated shellfish for different locations at

different time intervals during raining seasons                                                   136

4.14        The fat content of soaked and depurated shellfish for different locations at

different time intervals during dry seasons                                                         137                                                                                                                                               

4.15        The protein content of soaked and depurated shellfish for different

location at different time intervals during raining seasons                                                   139

                                                                                                                        

4.16        The protein content of soaked and depurated shellfish for different

locations at different time intervals during dry seasons                                           140                                                                                                                                                   

4.17        The Ash content of soaked and depurated shellfish for different location at

different time intervals during raining seasons                                                       142                                                                                                                                                            

4.18        The ash content of soaked and depurated shellfish for different locations at

different time intervals during dry seasons                                                             143

                                                                                                                                                                                                                                  

4.19        The crude fibre content of soaked and depurated shellfish for different

locations at different time intervals during raining seasons                                    145                                                                                       

4.20        The Crude fibre content of soaked and depurated shellfish for different

locations at different time intervals during dry season                                         146

                                                                                                                                                                                                        

4.21        The carbohydrate content of soaked and depurated shellfish for different

location at different time intervals during raining seasons                                    148

                                                                                                                                                                                                          

4.22        The Carbohydrate content of soaked and depurated shellfish for different

locations at different time intervals during dry seasons                                       149                                                                                                                                                                            

4.23        Variation in temperature of the water body at different locations in Azobie

creek during different month and seasons of the year                                           151

4.24        Variation in Dissolved oxygen of the water body at different locations in

Azobie creek during different month and seasons of the year                             154

4.25        Variation in pH of the water body at different locations in Azobie creek

during different month and seasons of the year                                                                         156

4.26        Variation in Nitrite content of the water body at different locations in

Azobie creek during different month and seasons of the year.                            158

4.27        Variation in Nitrate of the water body at different locations in Azobie creek

during different month and seasons of the year.                                                                       159

4.28        Variation in salinity content of the water body at different locations in

Azobie creek during different month and seasons of the year                         161

4.29        Variation in Amonia content of the water body at different locations in

Azobie creek during different month and seasons of the year                        163

4.30        Variation in Chloride content of the water body at different locations in

Azobie creek during different month and seasons of the year                        165

 






LIST OF PLATES

                                                                                            

3.1          Water Sample Collection from Azobie creek at monthly interval.                               50

3.2          Collection of the Sediment from Azobie creek                                                    50

3.3          Collection of the Oyster from the root of mangrove in Azobie creek        51

3.4          Collection of the Periwinkle from the sediment                                                 51

3.5          Depuration Stand                                                                                                                            53

3.6          Oyster shell                                                                                                                                        53

3.7          Oyster without shell                                                                                                                       54

3.8          Periwinkle in shell                                                                                                                            54

3.9          Periwinkle in the soaking tank                                                                                    55

3.10        Periwinkle in the depuration tank                                                                                            55

3.11        Oyster in the depuration tank                                                                                                    56

3.12        Oyster in the soaking tank                                                                                                           56

3.13        ZR Fungal/Bacterial DNA Kit                                                                                         75








 

CHAPTER 1

1.0  INTRODUCTION


1.1 BACKGROUND OF THE STUDY

Environmental pollution is  a major problem facing many settlements in Rivers State, especially Azobie creek where there was  recent oil vandalization., repairs of vessels, human faces, and increased in domestic wastes .The water  and the shellfish quality from this creek had been affected due to increase in environmental pollution. The increase in human population also in this area has led to increase in rate of pollution and contamination of most shellfish (Obodaiet al., 2010).

During soaking and depuration of shellfish harvested from a polluted area, the shellfish is dipped  in clean water in a container (unpolluted water) to allow the shellfish to cleanse or purge themselves by continuation of their normal filter-feeding and digestive process. This is a natural cleansing process that can be performed in a controlled environment by immersion in tanks of clean seawater to allow sewage contaminants to be purged (le Guyaderet al., 2006).

Depuration unlike cooking allows bivalve shellfish to be marketed as a live or fresh product. This is very common with species such as oyster, clams, periwinkles and mussels which are traditionally eaten live or are slightly cooked prior to consumption .It is the trust of this study to investigate the effect of depuration and soaking of live shellfish on heavy metal content, nutrient composition and microbial load of shellfish harvested from polluted creek of Rivers state.

Most water bodies in Rivers State are polluted. They receive indiscriminate effluent discharge of oil spill from heavily industrialized and highly populated Port-Harcourt metropolis.

 

These are some of the reasons why pollution occur frequently in RiverState. These also occur as a result of geological weathering, industrial processing of ore, metals, petroleum exploitation, vessel repair facilities, industrial wastes, oil spill and humanfaces(Chaerunet al., 2004;Muniaine Mujika et al., 2002;Loisyet al., 2005;Lees, 2000) These can impact negatively on the aquatic organisms, water and sediment. This causes most residents to eat contaminated seafoods.Clean-up of oil pollution in Rivers state is not frequent, and inadequate,leaving many people to cope with ongoing impacts of pollution to   humans (Chaerunet al.,2004).

 

Some shellfish such as oyster and periwinckle (Ifon and Umoh,2007) have the ability to accumulate  contaminants such as heavy metals, micro-organisms and polycyclic aromatic hydrocarbons in the environment. The ingestion of these contaminants by these organisms will not only affect the productivity and reproductive compatibility of these organisms but also affect the health of humans who depend on these organisms for food (Chaerunet al., 2004:Ifon and Umoh,2007)

 

The mangrove oysters (Crassostergasar are filter feeders, capable of accumulating micro-organisms in high concentration. They filter water at a rate of 2 to 5 litres/hour and also absorbs biotic and abiotic contaminant in their habitat (Nunes and Parsons,1998). They are one of the popular edible bivalves in Niger Delta. They have been identified as important vehicles of food-borne diseases because of their ability to absorb bacteria and viruses despite their nutritional benefit. These species may be eaten raw or lightly cooked and this increases the risk of food borne diseases.Shellfish like oysters feed by filtering large volumes of water and sediments. If the water contains microbiological and chemical contaminants and natural toxins, these will concentrate in their gut. As people normally eat shellfish raw, slightly cooked and without removing the gut, they are  likely to become ill if the shellfish  is harvested from heavily contaminated areas (Adeyerne, 2002). Oysters could be used as  vectors  for indicator of water quality since they harbor micro-organisms used as indicators of water quality One group of water and sewage indicators is the enteric bacteria,-fecal coliforms, which naturally inhabit the intestines of humans and are therefore used as surrogates in water quality assay (Nester et al., 2004).The range of bacteria  commonly found in waste water, tissues  and over flows discharges in a contaminated environment include Salmonella spp, Yersinia enterocolitica, Shigella spp Enterococcus spp, Escherichia coli, Clostridium perfringens, Staphylofcoccus spp, and Campylobacter spp (Scott, 2002) When enteric bacterium (Escherichia coli) is excreted into the water, they die at a slower rate than pathogenic bacteria such as Salmonella and Shigella and their presence might indicate presence of other pathogens in the water (Madigan et al., 2000).Periwinkle Tympanotonus Fuscatus is the most common gastropod in the Niger Delta Nigeria and are used for delicacy by most communities. They survive in a salinity range of 0.5 to 27.5ppt. They inhabit in stagnant waters where the substratum is muddy and rich in nutrients (Jamabo and Abowei, 2010).Tympanotonus  Fuscatus var fuscatus and Crastora gassar has been with high level of contaminant, and is the one of interest in this study (Davis et al.,2006).

 Some heavy metals that are discharged into aquatic ecosystems are likely collected as particles and accumulate in sediments and water (WHO,2006). A large reservoir of the metals in sediments exist and act as  overlying water column (WHO, 2006) and  lead to adverse ecologic effects and  danger to human health. The release of these contaminants from sediments and water may not only result from re-suspension of particulates, but also from the activities of microorganisms on the sediment and also on sediment-water interface. The extent of the risks of heavy metals and microorganisms is difficult to assess because of the complexity of biochemical activities that alter availability of metals in water, tissues and sediments. This is  why there  is need for effective depuration systems, to ensure  that  contaminant are washed out from the tissue (Obodai et al., 2010).

 Depuration is a process by which shellfish are placed in a clean water environment for a period of time to allow purging of its contaminants (Annon, 2006). Most researches and publications focus primarily on depuration of seafood rather than fresh water animals.  Modern seafood depuration, are segregated physical tanks using treated sea water sterilized either by chlorine, ultraviolet or Ozone.

 Depuration helps in preventing water and food borne diseases such as typhoid fever and other illness associated with sewage– borne bacteria and other contaminant (le Guyer, 2006). Depuration has been in practice globally within the last century but Nigeria has not recorded any depuration facilities, although the rate of consumption of these shellfish is increasing yearly in most parts of Nigeria. In research publications and government regulations, depuration is more embraced as “public protection” rather than as public awareness. In this study, depuration is embraced as both public protection and public awareness in some countries. It is now known that the success of depuration  depends on understanding of the environment, good monitoring practice, levels of pollution in the harvested area, and quality of water and shellfish  (FAO, 2008). Although depuration confer a level of additional food safety assurance to shellfish harvested  from areas which may be  highly polluted, it will not render shellfish grown in heavily polluted waters a safe product.  Literature shows that depuration successfully reduces bacteria, heavy metals and viral load in moderately polluted area. Depuration is capable of removing many bacterial species including faecal coliform from shellfish (Kelly et al., 1991). The efficacy of depuration is influenced by water temperature, salinity and turbidity which must be optimized to maintain the health status of shellfish (Annon, 2006). The success of depuration varies with species of shellfish, initial pathogen load, length of exposure to pathogens and pathogen distribution within shellfish tissues. It also depends on other factors such as mode of contamination and also the environmental conditions


1.2THE STATEMENT OF PROBLEM

Shellfish which constitute major source of protein to rural communities of River State is harvested from the stream, rivers and sea in this area. These water bodies are continually contaminated with highly polluted industrial waste heavily loaded with oil spill, heavy metals, human faeces, agricultural waste and several hydrocarbon polymers. The shellfish harvested from these water bodies are consumed without any treatment to reducethese contaminants. Consequently cases of food infection and intoxification relating to pathogenic microorganisms and heavy metals contaminants have been reported severally among these people. The major source of these food associated diseases could be from the sea food that they consume heavily. It became necessary to investigate the level and types of contaminants in the water bodies and the shellfish and the effect of depuration and soaking in reducing these contaminants in harvested    shellfish (periwinkle and oysters). This approach could be simple and affordable than checking the effluents in the water bodies within the depuration plan.


1.3       JUSTIFICATION OF THE STUDY

Shellfish, particularly Periwinkles and oysters are one of the major foods used for special delicacy in Rivers State. They are the cheapest and principal source of animal protein for the low-income group in the state. Periwinkles and oysters are of commercial value because most people use it in soups and as a delicacy. Most resort to shellfish as a healthy alternative to meat today because they are cheaper, provides high quality proteins and good profile of fatty acids and easily digestible mineral salts (Andrew, 2001).

Unfortunately, shellfish grow naturally in coastal waters and are exposed toseries of contaminants including pathogenic enteric micro-organisms, cyclic hydrocarbons and toxic heavy metals. Thus microbial quality and levels of heavy metals and polycyclic hydrocarbon content of the inhabiting water body determine safety of the shellfish (Offen et al., 2009). River state is highly industrialized in Nigeria with discharge of oil spill, human faeces, heavy metals contaminants, cyclic hydrocarbons, mining residues, other mineral pollutants in the water bodies where the fishes inhabit (Koopmans et al., 2002). Several cases of food poisoning and untimely death of eating contaminated shell fish have been reported in this area (Davis et al., 2006; Chaerun et al., 2004; Jackson and Ogburum, 1996; Otsu, 1999; Bahir et al., 2012)

Many communities in River State are affected and people in these communities are reluctant and scared to eat locally caught shellfish. The people have no idea of any method to decontaminant their shellfish before consumption .Again, they sell their fish to buyers  in the same level of contamination. Depuration and soaking is one of the simplest  cost effective approach that processors can use to make shellfish safe for consumers .The people are unaware of this approach. Depuration in this study could be used to produce natural safe shellfish to ensure consumer safety and sustainable supply of high quality flesh protein to the common man in Rivers state and its environment. This will improve food security.


1.3       OBJECTIVE OF THE STUDY

1.3.1 General objective of the study

The main objective is to investigate the effect of different time intervals of soaking and depuration on microbial load, heavy metal, nutritional composition of shellfish harvested from polluted creek in rivers state at different seasons


      1.3.2        The specific objectives of the study are to:

      1.      To construct depuration system

      2.      To  determine the concentration of heavy metal present in the harvested periwinkle and oyster

      3.      To evaluate the microbial quality and proximate composition of the harvested periwinkle and oyster

      4.      To determine the effect of depuration and soaking time on heavy metals load on periwinkle and oyster harvested at different location from a polluted creeks of Rivers state, Nigeria at different seasons.

      5.      To determine the effect of soaking and depuration time on nutritional composition of periwinkle and oyster harvested at different location from a polluted creeks of  Rivers state, Nigeria at different seasons

    6.      To determine the effect of depuration and soaking time on microbial quality of periwinkle and oyster harvested from different location from a polluted creeks of Rivers state, Nigeria at different seasons .

    7.      To evaluate the variation in physical and chemical parameters of the shellfish habitat at different location of the Azobie Creek.



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