in vitro culture and isoenzyme analysis of giardia lamblia

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Phillips RE, Boreham PFL, Shepherd RW. 1984. Cryopreservation of viable Giardia intestinalis trophozoites. Transactions of the Royal Society of Tropical Medicine and Hygiene. 78: 604-606. Polis MA, Tuazon CU, Ailing OW, Talmanis E. 1986. Transmission of Giardia lamblia from a day care center to the community. American Journal of Public Health. 76: (9) 1142 -1144. Proctor EM, Isaac-Renton JL, Boyd J, Wong 0, Bowie WR. 1989. Isoenzyme analysis of human and animal isolates of Giardia duodenalis from British Columbia, Canada. American Journal ofTropical Medicine and Hygiene.41:(4) 411-415. I Rabbani GH, Islam A. 1994. Giardiasis in humans: Populations most at risk and prospects for control for control.lrL Thompson RCA, Reynoldson JA, Lymbery AJ. (eds). Giardia: From Molecules to Disease. Wallingford, UK; CAB International. pp 217-249. Rendtorff RC.1954. The experimental transmission of human intestinal protozoan parasites. 11. Giardia lamblia cysts given in capsules. American Journal of Hygiene. 59: 209-220. Rice EW and Schaefer FW111. 1981. Improved in vitro excystation procedure for Giardia lamblia cysts. Journal of Clinical Microbiology. 14: (6) 709-710. RitChie LS. 1948. Ether sedimentation technique for routine stool examination. Bulletin of the Us Army Medical Department. 8: 326. Ritchie LS, Pan C, Hunter GW.1952. A comparison of the zinc sulfate and the MGL (formalin-ether) technics. Journal of Parasitology 38:(4) Suppl. 16. Roberts-Thompson IC. 1993. Genetic studies of human and murine giardiasis. Clinical Infectious Diseases. 16 (SuppI.2): S98-104. Roberts-Thompson IC, Mitchel GF, Anders RF, Tait BD, Kerlin P, Kerr-Grant A, Cavanagh P. 1980. Genetic studies in human and murine giardiasis. Gut. 21: 3970- 401. Roberts-Thompson IC, Stevens OP, Mahmoud AF, Warren KS. 1976. Giardiasis in the mouse: An animal model. Gastroenterology. 71 :57 -61. Ryder RW, Reeves WC, Sack RB. 1985. Risk factors for fatal childhood 'diarrhoea: a case control study from two remote Panamanian islands. American Journal of Epidemiology. 121: (4) 605-610. Saha TK, Ghosh TK.1977. Invasion of small intestinal mucosa by Giardia lamblia in man. Gastroenterology. 72: 402-405. Sargeaunt PG, Wi"iams JE, Jackson TFHG, Simjee AE. 1982. A zymodeme study 165
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IN VITRO CULTURE AND ISOENZYME ANAL
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(j)edicated to my {ate parents :No
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the positive specimens by sucrose g
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Declaration This study represents o
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ACKNOWLEDGEMENTS The author wishes
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Table of Contents Page CHAPTER 1 -
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4.2.1. Animal Sourcing and Care ...
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APPENDIX 8 List of enzyme systems u
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List of Tables Page Table 1.1 Commo
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List of Plates Page Plate 2.1 An eo
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ABBREVIATIONS AND UNITS OF MEASUREM
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agi/is in 1882. In 1914, Alexeieff
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unique about Giardia in humans, whi
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To-date, use of the different speci
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Differentiation into the cyst stage
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1.3 Transmission 1.3.1 Human hosts
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Owen, 1984) 1.3. 2 The Zoonosis of
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1.4 Distribution The morbidity and
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children by Schutte et al. in 1981
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of (a) the enormous absorptive capa
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Ig M (100% of patients), IgG (70%),
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chymotrypsin) between different iso
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impact on the physical growth of 29
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Figure 1.3. Transmission Electron m
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among children between the ages of
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where immunocompromised mice have p
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up to 4 distinct genotypes within e
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in initiation of infection (excysta
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In order to address these and many
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multiple stool examinations in 4 of
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2.2 MATERIALS AND METHODS 2.2.1 Spe
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Table 2.1. Semi-quantitative assess
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2.2.3.2 The modified Zinc sulphate
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Preliminary work with five samples
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% Viability = unstained cysts/total
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Table 2.3. Prevalence of Giardia in
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Table 2. 4. Monthly summary of stoo
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spp (59%). Other parasites less fre
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Observations • A large proportion
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Plate 2.1. An eosin-stained prepara
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2.4 DISCUSSION In terms of yield, i
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examination of wet preparations. Th
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ecognised (on unstained preparation
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cultivation in vitro (Kasprzak & Ma
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3.2 MATERIALS AND METHODS Three exc
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3.2.3 The modified Acid Pepsin meth
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• After 15 minutes incubation: Ma
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Plate 3.2 (b) A completely excysted
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Twenty-four batches of cysts (71 %
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3.3.3 The modified Acid Pepsin excy
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3.4 DISCUSSION Since our source of
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an unreliable indicator of viabilit
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propagating Giardia for subsequent
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4.2 MATERIALS AND METHODS 4.2.1. An
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A litter comprising of 7 C57BU6 mic
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for attached trophozoites and maint
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days pi. However, accurate quantifi
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Table 4.4. Summary of the inoculati
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not be infected in comparison with
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A proportionately higher percentage
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These findings strongly suggest tha
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CHAPTERS IN VITRO CULTURE OF GIARDI
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Diamond et al.'s (1978) medium. He
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Majewska, 1985) United States (Nash
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were evaluated. 5,2,1,2 Biosate Bio
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Table 5.1 Outlines a relative semi-
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dislodged by chilling the culture t
Page 135 and 136: Table 5.2. Sensitivity to Ciproflox
Page 137 and 138: Table 5.3. Summary results of in vi
Page 139 and 140: Once established in culture for a l
Page 141 and 142: Plates 5.3 and 5.4 Confluent growth
Page 143 and 144: difficult to produce viable culture
Page 145 and 146: dominance of certain genotypes unde
Page 147 and 148: Cryopreservation and successful ret
Page 149 and 150: adjusted to 5°C per minute from am
Page 151 and 152: 6.3 RESULTS Trophozoites have been
Page 153 and 154: Table 6.1. A longevity record of sa
Page 155 and 156: Diamond (1995) stated that an impor
Page 157 and 158: into two categories. One group cont
Page 159 and 160: technique to the first local (South
Page 161 and 162: for Giardia Iysates. These included
Page 163 and 164: 4. To exclude the presence of bacte
Page 165 and 166: Plate 7.1 Banding pattern of 8 seri
Page 167 and 168: PGM activity was displayed by Giard
Page 169 and 170: Plate 7.6. Hexokinase Lane 1. Unino
Page 171 and 172: Plate 7.8. Phosphogluconate dehydro
Page 173 and 174: stably expressed between different
Page 175 and 176: techniques, which would allow diffe
Page 177 and 178: Belosevic M, Faubert GM, MacLean JD
Page 179 and 180: Farthing MJG, Mata L, Urrutia JJ, K
Page 181 and 182: Heidelberg pp.397-398. Hiatt RA, Ma
Page 183 and 184: patients with X-linked agammaglobul
Page 185: Press, Calgary. pp 57-58. Nash TE,
Page 189 and 190: Wolfe M. S.1979. Giardiasis. Sympos
Page 191 and 192: APPENDICES APPENDIX 1 Information t
Page 193 and 194: Unayo imvume yokunqaba ukungenela l
Page 195 and 196: Appendix 3 Excystation and culture
Page 197 and 198: Appendix 4 Results of excystation a
Page 199 and 200: Appendix 5 Results of in vitro (aci
Page 201 and 202: Key to Appendix 5 *Replicated inocu
Page 203 and 204: Appendix 7 Preparation of culture m
Page 205 and 206: Appendix 8 List of enzyme systems u
Page 207 and 208: Appendix 9 Appendix (9a) Preparatio
Page 209: Appendix 10. Quantitative descripti
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