ISOLATION AND IDENTIFICATION OF MICROBIAL PATHOGENS ASSOCIATED WITH THE EXTERNAL BODY SURFACE OF PERIPLANETA AMERICANA IN UMUAHIA, ABIA STATE.

                ABSTRACT

This study was carried out to isolate and identify the microbial pathogens on the external body surfaces of Periplaneta americana collected from toilet areas, residential/hostel areas and health facilities in Umuahia, Abia State. A total of 100 cockroaches from toilets (30), residential/hostel areas (46) and health facilities (24) in Umuahia were collected and their external body parts screened for bacterial, fungal and parasitological carriage. Escherichia coli (20.00%) and Klebsiella species (17.14%) were the most common bacteria in the cockroaches analyzed while, Staphylococcus aureus (21.74%) assumed rather high values in those gotten from health facilities. Aspergillus species (33.16%) was the most prevalent fungal group. Quantitative analysis of the total viable count and microbial loads of the bacterial and fungal isolates from the three sites revealed that the isolates obtained from toilet areas were high as compared to those from residential areas and health facilities. Parasitological investigations revealed the preponderance of Ascaris lumbricoides (32%) in all the sampled areas. Antibiotics susceptibility of all the bacterial organisms isolated showed that those obtained from health facilities were highly resistant to the antibiotics used as compared to those from toilet and residential/toilet areas. This result indicated that Periplaneta americana is a possible reservoir of several pathogens via their bodies and thus may spread multiple drug resistant species.

TABLE OF CONTENTS

TITLE PAGE i

CERTIFICATION ii

DEDICATION iii

ACKNOWLEDGEMENT iv

TABLE OF CONTENT v

LIST OF TABLES vi    

ABSTRACT vii

                                                      CHAPTER ONE

1.0       INTRODUCTION 1-3                                                                                                

1.1 BACKGROUND OF STUDY 3

1.2 RATIONALE OF STUDY 3

1.3 AIM AND OBJECTIVES OF THE STUDY 4

1.4 SIGNIFICANCE OF THE STUDY 4

1.5 LIMITATIONS OF THE STUDY 4

                                                     CHAPTER TWO

2.0 LITERATURE REVIEW 5

2.1 HABITAT 5

2.2 FEEDING 5

2.3 IDENTIFICATION AND CHARACTERIZATION 5-6

2.4 COCKROACHES AND DISEASE TRANSMISSION 6-8

                                                   CHAPTER THREE

3.0      MATERIALS AND METHODS 9

3.1      STUDY AREA 9

3.2      MATERIALS 9-10

3.3      CAPTURE AND COLLECTION OF COCKROACHES 10

3.4 PROCESSING OF COCKROACHES 11

3.5      MEDIA PREPARATION 11

3.6      IDENTIFICATION OF PARASITES 11

3.7 QUANTITATIVE ESTIMINATION OF BACTERIAL AND FUNGAL

            ISOLATES 12

3.8 ISOLATION AND IDENTIFICATION OF FUNGAL

            ISOLATES 12

3.9 ISOLATION AND IDENTIFICATION OF BACTERIAL

           ISOLATE 12-13

3.9.1    GRAM STAIN 13

3.9.2    CATALASE TEST 14

3.9.3 COAGULASE TEST 14

3.9.4 CITRATE TEST 14

3.9.5 MOTILITY, INDOLE, UREASE TEST 14-15

3.9.6 TRIPLE SUGAR IRON (TSI) TEST 15

3.10 ANTIBOTIC SUSCEPTIBILITY TESTING 16

3.11 DATA ANALYSIS 16

                                               CHAPTER FOUR

4.0 RESULT 17-30

 CHAPTER FIVE

5.0 DISCUSSION, CONCLUSSION AND RECOMMENDATION 31

5.1 DISCUSSION 31-33

5.2 CONCLUSION 33

5.3 RECOMMENDATION 33

REFFERENCES

                                                  LIST OF TABLES

                                        CHAPTER ONE

1.0 INTRODUCTION

Cockroaches are primitive highly successful winged insects. They are considered one of the most adaptable insect groups ever in human history (Etim et al., 2013). Evidence of their continued coexistence with people throughout history is testimony to how adaptable some cockroaches are to the habits of people (Burgress, 1981). They are born inebriates and omnivorous scavengers in nature and have been noted by scientists to have survived more than 300 million years (Zurek and Schal, 2004). Ancient fossils had the same appearance as today’s cockroaches: oval and flat with long legs and antennae.

Of the over 4, 000 known species of cockroaches, only a dozen can be considered as pests. Syananthropic animal species, including cockroaches, live in most human habitations, especially where food is stored, processed, prepared, or served (Taha et al., 2014). They breed in buildings and share human food and shelter. Apart from human food, they also consume putrefied and decaying organic matter (Jeffery et al., 2012, Bennet, 2008 and Pavlik and Faklinham, 2009), and thus are easily, sited crawling in and/ or near broken several lines or refuse dump sites in major towns and cities around the world. During the day, these insects hide in isolated locations or in gaps of the walls, but become active at night when they can move about unnoticed (Milner and Pereira, 2007, Bonnefoy et al., 2008 and Lihoreau et al., 2012). The American cockroach, Periplaneta americana is by far the most serious and predominant metropolitan pest in Umuahia and  her environs judging by how often they are commonly sited in large numbers in  many households, kitchens/restaurants, offices, grocery stores, broken sewage and refuse disposal sites in and around Umuahia metropolis.

The medical importance of cockroaches is much greater than generally realized as they have been shown to harbor a number of pathogenic and non-pathogenic microorganisms (Fotedar et al., 1991). Cockroaches are known vectors of human entero-pathogens as recent research findings have provided substantial evidence of the isolation of various human pathogens from these insects (Akintola et al., 2013, Kausar et al., 2013, Tachbele et al., 2006 and Hossein et al., 2003). Cotton et al,. suggested that cockroaches are possible vectors of Klebsiella pneumonia in the hospital environment. In a related study of bronchial asthma by Tsai and Chen, 40% of patients in China were found to be allergic to German cockroaches (Tsai and Chen, 1999). Furthermore, a comparative analysis of pathogenic organisms in cockroaches from different socio-economic community settings in Edo State, Nigeria showed that organisms such as Bacillus spp, Escherichia  coli, Proteus mirabilis, Proteus vulgaris, Pseudomonas aeruginosa, Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Citrobacter freundi and Salmonella spp can be harbored on the body surface and alimentary tract of cockroaches gotten from the study area (Clement et al., 2014). In the same study, Clement et al., reported the isolation and identification of some fungal organisms such as Aspergillus spp, Mucor spp, Saccharomyces cerevisiae and a few parasites which included Ascaris lumbricoides, Trichuris trichiura, Coccidia and Entamoeba histolytica (Clement et al., 2014).

Indeed, numerous published papers recognized the association of cockroaches with several infectious diseases and spread of pathogenic and drug resistant microbes such as Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Klebsiella spp, Salmonella typhi and even some fungi, viruses, protozoa and Helminthes (Taha et al., 2014). However, direct evidence that indicates that cockroaches could be involved in the transmission of bacterial diseases is largely circumstantial as they are merely passive vectors which only carry bacteria that are already present in the environment (Akintola et al., 2013). Hence, their role in the direct transmission of infection has seldom been established and is somewhat uncertain (Tatfeng et al., 2005 and Marriott et al., 2006).

1.1 BACKGROUND OF STUDY

Cockroaches are a major health hazard in both urban and rural settings. Cockroaches are among the most common pests in public dwellings and health facilities worldwide. Their presence can raise safety concerns, especially as they may be carriers of pathogenic organisms. The American cockroach (Periplaneta americana) harbor and disseminate about 150 pathogenic and non-pathogenic microorganisms. They play important roles in the transmission of different diseases either mechanically or biologically in health centres hospitals, domestic dwellings e.t.c. around and within the Umuahia metropolis.

1.2 RATIONALE OF STUDY

Free wandering movements of cockroaches from one location to another coupled with their possible contribution to disease transmission as clearly seen by the association of cockroaches with several infectious diseases of man and animals, spread of pathogenic and drug resistant microbes prompted this study.

1.3 AIM AND OBJECTIVES OF THE STUDY

The aim of this study is to evaluate the prevalence of various pathogenic microbial flora associated with the external body surface of the American cockroaches (Periplaneta americana) from different locations within the Umuahia capital city. The objectives are;

v To isolate and identify the various microbial species (bacteria, fungi and parasites) carried on the external body surface of American cockroaches.

v To enumerate their fungal and bacterial loads as well to determine their parasite density.

v To ascertain the antibiotic susceptibility pattern of the bacterial isolates.

1.4 SIGNIFICANCE OF THE STUDY

This research work would be of great importance to both the Government and the general public most especially the Umuahia people and their neighbors by sensitizing and enlightening them on the inherent zoonotic and nosocomial effects/roles of the American cockroach (Periplaneta americana) in their homes, offices, restaurants, health facilities etc. with a view to improving or putting in place their infection control unit.

1.5 LIMITATIONS OF THE STUDY

The major stumbling block that sought to mar the effective carrying out of this study was that of finance and time. The study operated with a very small budget and lasted for only 4 weeks. Huge sums of money and a longer period of time were needed to fully study larger samples sizes. Also, the unavailability of typing sera which would have aided immensely in the determination of specific serotypes of E. coli with disease causing abilities posed a major setback, as this would have been very useful in our research.

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