Multiple Antimicrobial Resistances of Bacteria Isolated from Samples of Recreational Beach in ...

167 Pages·2015·3.16 MB·English

by  Maya Mohammed

Islamic University - Gaza –
ةزــــــغ ةيملاـــــسلإا ةــــعمالجا
Deanship of Graduate Studies ايلعلا تاساردلاو يملعلا ثحبلا نوؤش
Faculty of Science موـــــــــــــــــلعلا ةـــــــــــــــيلك
Environmental Sciences Master ةيئيبلا مولعلا يرتــــشجام جـــمانرب
Programme/Environmental Health ةـــــــــــــيئيبلا ةـــــــــــــــــحصلا

Multiple Antimicrobial Resistances of Bacteria
Isolated from Samples of Recreational
Beach in Gaza Strip
ةلوزعلما اييرتكبلا لبق نم ةيويلحا تاداضملل ةددعتلما ةمواقلما
ةيمامجتسلاا ةزغ ئطاوش نم تانيع نم

By
Fatema Mohammed Elfarra

Supervised by

Prof. Dr. Abdelraouf A. Elmanama Dr. Kamal Elnabris
Prof. of Microbiology Associate prof. of Marine Bio Environmental

A Thesis Submitted in Partial Fulfillment of the Requirement for the Degree
Master of Science in Environmental Sciences
The Islamic University – Gaza – Palestine

2015 AD- 1436 AH

ميحرلا نحمرلا للها مسب

In the Name of Allah

نْ َأوَ يَّ دَِلاوَ ىَلعَوَ يَّ َلعَ تَ مْ عَ ْنَأ يِتَّلا كَ َتمَعِْن رَكُ شْ َأ نْ َأ يِنعْزِوَْأ بِّ رَ}
ِ ِ ِ ِ ِ ِ ِ ِ ِ ِ
{نيحلاصَّ لا كَ داَبع يف كَ تمَحْرَب ينلْخدَْأوَ ُهاضَ رْ َت احً لاصَ لَ مَعْ َأ
)91 :ةيلآا لمنلا)

“O my Lord! So order me that I may be grateful for Thy
favors’ which Thou have bestowed on me and on my
parents, and that I may work the righteousness that will
please Thee: and admit me, by Thy Grace, to the ranks of
Thy Righteous Servants”

I
Dedication

I dedicate this study to the soul of my beloved
father, who always wished to see me standing here
today, to the kindest heart ever, to my mother for
her endless prayers for me to fulfill my ambition.

With my respect and love
Fatema El Farra

II
Acknowledgment

I fully praise Allah for all the blessings he grants us, I also deeply thank those ever
encouraged me so finally I had my dream turned to be a bright fact.

My special appreciation is for Prof. Dr. Abdelraouf Elmanama for his endless
patience, guidance, support and valuable pieces of advice, which deeply enriched my
research.

I can never forget Dr. Kamal Elnabris rich suggestions that paved the way for my
study to see the light.

Deepest thanks for the staff of Environmental Science- post graduate program in the
Environment and Earth Sciences Department at the Islamic University, for their
academic and scientific supervision throughout my study.

To all staff at the public health laboratory-Ministry of health for all the help in
doing the sample analysis and full access to their facilities that they provided me.
To the Environmental and Rural research center- IUG for assisting me in sampling.
This work would not have been possible without the extensive field Mr. Abdel
Kareem Abu Hatab for helping me in sampling.

Lastly, I offer my regards and respect to all those who ever supported me in any aspect
during the completion of my project.

III
Abstract
The aim of the present study is a document the occurrence of pathogenic bacteria in
seawater, beach sand and mussel samples collected from five locations along the coast
of the Gaza strip and to determine the extent that these bacteria possess multi-drug
resistance. Pathogens were isolated and identified by microbiological and biochemical
methods and their susceptibility was determined against different antimicrobial agents
by using the agar disc diffusion method and according to NCCLS (National Committee
for Clinical and Laboratory Standard) guidelines. In most locations, cultivable fecal
indicator bacteria such as fecal coliform (FC) and fecal streptococci (FS) were more
abundant in the sand (an average of 846.6 cfu/100 g) than in the sea water (579.5
cfu/100ml) and the number of FS in sand samples were higher than that of FC.
Population density of both FC and FS was generally higher at Khan Yunis, South of
Wadi Gaza outlet and North of Wadi Gaza outlet. Mussels were found to accumulate
more FS than FC. The results also indicated that all sites comply with mandatory
standards for water quality in the directive, and only the northern side of Wadi Gaza
outlet do not achieve the more stringent guideline values for both FC and FS in water.
Conventional biochemical tests identified 411 isolates belong to four bacterial groups of
Enterobacteriaceae (42.4%), Enterococci (17.6%), Pseudomonas aeruginosa (23.7%)
and Staphylococci (16.3%). Enterobacteriaceae, Enterococci, Pseudomonas aeruginosa
and Staphylococci isolates were tested for their susceptibility to 10, 6, 8 and 10
antimicrobials respectively. The incidence of multiple resistances for the four bacterial
groups from the different sample types (mussels, seawater and beach sand) and from the
different sampling locations are presented. Amongst the Enterobacteriaceae isolates,
94.3% were resistant to at least one antimicrobial agent and 80.5% exhibited
simultaneous resistance to more than one agent i.e., they are Multiple Antimicrobial
Resistance (MAR). It also found that 100% of Enterococci isolates were resistant at
least to one antimicrobial and 89.4% were MAR. For Pseudomonas aeruginosa isolates,
68.5% were resistant at least one agent and 43.8% were MAR. Staphylococci isolates
showed that 98.4% were resistant to at least one agent and 88.6% were MAR. The
greatest frequency of resistant in all Enterobacteriaceae isolates was found against
Tetracycline (67.9%) and Amoxicillin-clavulanic (56%). For the Enterococci, the
highest resistance was against Vancomycin (84.8%) and Ampicillin (80.3%).
IV
Pseudomonas aeruginosa highest resistant was against Piperacillin (55.1%). The
highest resistance of Staphylococci isolates was against Penicillin )91.8%(,
Vancomycin )68.9%( and Oxacillin )62.3%(. All the calculated values of MAR index
were higher than 0.2 demonstrating that the area of study is considered a high risk
source of contamination environment where antimicrobial agents use are common. In
conclusion, the results of the present study shows the widespread occurrence of
antibiotic resistant bacteria in all types of samples tested and from all locations along
the coast of the Gaza strip and their presence may become a potential human health
hazard. Consequently efforts should be exerted to minimize the discharge of untreated
or partially treated sewage from both domestic and hospitals sources which represent
the prime source of antimicrobial agents in the marine environment in this area.

Keywords: Antimicrobial resistance, multiple antimicrobial resistances, Enterobacteriaceae,
Gaza beach, Enterococci, Staphylococci, Pseudomonas aeruginosa

V
Abstract in Arabic language
صخلملا
يبِسٚ شحجٌا ٖبٍِ ًف خٍثٚشىٌٍّا دادبضٌٍّ خِٚبمٌّا خضشٌّّا بٌشٍزىجٌا دٛجٚ كٍثٛر ىٌإ خساسذٌا ٖز٘ فذٙر
ٖز٘ نلازِا خجسد ذٌذحرٚ حضغ عبطل ًحبس يٛط ىٍػ غلاِٛ خسّخ ِٓ بٙؼّج ُر ًزٌا شحجٌا حٍث دبٍٕػٚ ئطبشٌا
خٍئبٍٍّىٌاٚ خٍجٌٍٛٛثٚشىٌٍّا قشطٌبث بٙفٌشؼرٚ خضشٌّّا بٌشٍزىجٌا يضػ ُر .خٌٚدلأٌ حدذؼزٌّا خِٚبمٌٍّ بٌشٍزىجٌا
بمفٚٚ صاشللأا ششٔ خمٌشط َاذخزسبث خٍثٚشىٌٍّا دادبضٌّا ِٓ خفٍزخِ خػّٛجٌّ بٙزِٚبمِ ذٌذحر ُر هٌزوٚ خٌٍٛحٌا
خٌصاشجٌا ٌْٛٛمٌا بٌشزىث ذٔبو غلاٌّٛا ُظؼِ ًف .(NCCLS( خٌشجخٌّاٚ خٌشٌشسٌا دبسبٍمٌٍ خٍٕطٌٛا خٕجٌٍا شٌٍبؼٌّ
cfu( شحجٌا ٖبٍِ ًف بِٕٙ )ُج100/ 846.6 cfu ظسٛزّث( يبِشٌا ًف حشفٚ شثوأ (FS) خٌصاشجٌا دبٌذمؼٌاٚ (FC)
ًف ىٍػأ FS ٚ FC ِٓ لاو خفبثو ذٔبوٚ .FC ِٓ ىٍػأ ًِشٌا دبٍٕػ ًفFS دذػ ْبوٚ (ًِ100/ 579.5
ًف خّواشزِ FSٚ FC ِٓ شثوأ داذػأ ىٍػ سٛثؼٌا ُر .حضغ يداٚ يبّشٚ حضغ يداٚ ةٕٛجٚ سٌٔٛ ْبخ كطبِٕ
اذػ بِ غلاٌّٛا غٍّج ْأٚ ،ٖبٌٍّا حدٛجٌ خٍِاضٌلإا شٌٍبؼٌّا غِ كفزر غلاٌّٛا غٍّج ْأ جئبزٌٕا دسبشأ بّو .شحجٌا حٍث
.FS ٚ FC ِٓ ًىٌ ٖبٌٍّا حدٛجٌ خِاشص شثولأا خٍٍٙجٛزٌا شٌٍبؼٌّا كمحر حضغ يداٚ ِٓ ًٌبّشٌا تٔبجٌا
،خٌٛؼِ خٌشٍزىث %42.4 ًٌبزٌبو ذٔبو دبػّٛجِ غثسأ ىٌإ ًّزٕر خٌضػ 411 ددذح خٌٍٛحٌا خٍئبٍٍّىٌا داسبجزخلاا
خٌٛؼٌّا بٌشٍزىجٌا خِٚبمِ سبجزخا ُر .خٌدٛمٕػ داسٛىِ %16.3ٚ خٌسبجٔص خفئاص %23.7 ،خٌٛؼِ داسٛىِ %17.6
خٍثٚشىٍِ دادبضِ 8 ي خٌسبجٔضٌا خفئاضٌا ، خٍثٚشىٍِ دادبضِ6 ي خٌٛؼٌّا داسٛىٌّا ،خٍثٚشىٍِ دادبضِ10 ي
خسّخٌا ِٓ خٌٚضؼٌّا خٍثٚشىٌٍّا دادبضٌّا خِٚبمِ جئبزٔ دشٙظأ .خٍثٚشىٍِ دادبضِ10 ي خٌدٛمٕؼٌا داسٛىٌّاٚ
٪80.5 ْأٚ ًللأا ىٍػ دبثٚشىٌٍّا دادبضِ ِٓ ذحاٌٛ خِٚبمِ خٌٛؼٌّا بٌشٍزىجٌا ِٓ ٪94.3 ْأ خفٍزخٌّا غلاِٛ
ذٔبو خٌٛؼٌّا داسٛىٌّا دلاضػ ِٓ ٪100 ْأ ذجٚ بّو .دبثٚشىٌٍّا دادبضِ ِٓ شثوأ ٚأ ٍٕٓثلا خِٚبمِ ذٔبو
دلاضػ ِٓ ٪68.5 ذٔبوٚ .ًثٚشىٍِ دبضِ ِٓ شثولأ خِٚبمِ ٪89.4 ٚ ًللأا ىٍػ ذحاٚ ًثٚشىٍِ دبضٌّ خِٚبمِ
دشٙظأٚ .ًثٚشىٍِ دبضِ ِٓ شثولأ خِٚبمِ ٪43.8 ٚ ًللأا ىٍػ ذحاٚ ًثٚشىٍِ دبضٌّ خِٚبمِ خٌسبجٔضٌا خفئاضٌا
دار ذٔبو ٪88.6 ٚ ًللأا ىٍػ ذحاٚ ًثٚشىٍِ دبضٌّ خِٚبمِ ذٔبو خٌدٛمٕؼٌا داسٛىٌّا ِٓ %98.4 ْأ دلاضؼٌا
هٍٔلاٛفلاىٌا ٍٍٍٓسٍسوِٛأٚ )٪67.9( ٍٍٓىٍساشززٌا ذض خِٚبمِ ىٍػأ خٌٛؼٌّا بٌشٍزىجٌا دشٙظأٚ .حدذؼزِ خِٚبمِ
دشٙظأٚ .)٪80.3( ٍٍٓسٍجِلأاٚ )٪84.8( ٍٓسٍِٛىٔبف ذض خِٚبمِ ىٍػأ خٌٛؼٌّا داسٛىٌّا دشٙظأ .)٪56(
خِٚبمِ ىٍػأ خٌدٛمٕؼٌا داسٛىٌّا دلاضػ دشٙظأ بٍّٕث .)٪55.1( ٍٍٍٓساشٍجٍث ذض خِٚبمِ ىٍػأ خٌسبجٔضٌا خفئاضٌا
دذؼزٌّا خِٚبمٌّا ششؤِ ْبو ذمٌٚ .)٪62.3( ٍٍٍٓسبسوٚأٚ ،)٪68.9( ٍٓسٍِٛىٔبفٚ ،)٪91.8( ٍٍٓسٕجٌا ذض
غلاٌّٛا غٍّج ْأ ىٍػ يذٌ بِّ ب٘سبجزخا ُر ًزٌا غلاٌّٛا غٍّج ًف 0.2 ِٓ ىٍػأ )MARI( خٍثٚشىٌٍّا دادبضٌٍّ
.ٍُثاشجٌا دادبضٌّ غئبشٌا َاذخزسلاا تجسث خثٍِٛ شجزؼر خسٚسذٌّا
عاٛٔأ ًو ًف خٍثٚشىٌٍّا دادبضٌٍّ خِٚبمٌّا بٌشٍزىجٌٍ غساٚ سبشزٔا شٙظر خساسذٌا ٖز٘ جئبزٔ شٙظر ،َبزخٌا ًفٚ
ًىشٌ ْأ ٓىٌّّا ِٓ بٌشٍزىجٌا ٖز٘ دٛجٚ ْأٚ حضغ عبطل ًحبس يٛط ىٍػ غلاٌّٛا ًو ِٓ ب٘سبجزخا ُر ًزٌا دبٍٕؼٌا
شٍغٌا ًحصٌا فشصٌا ٖبٍِ فٌشصر ًٍٍمزٌ دٛٙجٌا يزث تجٌ ٗٔئف هٌر ىٍػ ءبٕثٚ .ْبسٔلإا خحص ىٍػ اشطخ
VI
خٍثٚشىٌٍّا دادبضٌٍّ ًسٍئشٌا سذصٌّا ًثّر ًزٌاٚ دبٍفشزسٌّاٚ خٍٍحٌّا سدبصٌّا ِٓ بٍئضج خجٌبؼٌّا ٚأ خجٌبؼٌّا
.خٌشحجٌا خئٍجٌا ًف
ئطبش ،خٌٛؼٌّا بٌشٍزىجٌا ،خٍثٚشىٌٍّا دادبضٌٍّ حدذؼزٌّا خِٚبمٌّاٚ ،خٍثٚشىٌٍّا دادبضٌّا خِٚبمِ :ةيحاتفم تاملك
.خٌسبجٔضٌا خفئاضٌا ،خٌدٛمٕؼٌا داسٛىٌّا ،خٌٛؼٌّا داسٛىٌّا ،حضغ

VII