in vitro culture and isoenzyme analysis of giardia lamblia

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Gilman RH, Brown KH, Visvesvara GS, Mondal G, Greenberg, Sack RB, Brandt F, Khan MU. 1985. Epidemiology and serology of Giardia lamblia in a developing country: Bangladesh. Transactions of the Royal Society of Tropical Medicine and Hygiene. 79,469-473. Gilman RH, Marquis GS, Miranda E, Vestegui M, Martinez H. 1988. Rapid reinfection by Giardia lamblia after treatment in a hyperendemic third world community. Lancet. i, 343-345. Gordts B, Hemelhof W, van Tilborgh K, Retore P, Cadranel S, Butzler JP. 1985. Evaluation of a new method for routine in vitro cultivation of Giardia lamblia from human dodenal fluid. Journal of Clinical Microbiology. 22, (5) 702-704. Gordts B, Retore P, Cadranel S, Hemelhof W, Rahman M, Butzler JP. 1984. Routine culture of Giardia lamblia trophozoites from human duodenal aspirates. The Lancet 2:137-138 . Hall A. 1994. Giardia infections: Epidemiology and nutritional consequences. in Thompson RCA, Reynoldson JA, Lymbery AJ. (eds).Giardia: From molecules to disease. Wallingford, UK; CAB International.pp 251- 2291. Hamilton HO and Jackson TFHG. 1990. Efficacy of a small water treatment method for inactivating Giardia cysts in raw water. Water SA. 16, (3) 201-204. Ha'rris H, Hopkinson OA. 1976. Handbook Of Enzyme Electrophoresis in Human Genetics. North Holland. Hass J, Bucken EW. 1967. Zum Krnakheeiitwert der Lambien Infektion. Deutshe Medizzinissche Wochensssscrift 92, 1869-1871. (Abstract). Hautus MA, Kortbeek LM, Vetter JC, Laarman JJ. 1988. In vitro excystation and subsequent axenic growth of Giardia lamblia. Transactions of The Royal Society Of Tropical Medicine And Hygiene. 823, (6): 858-861 . Hegner RW. 1925. Excystation in Giardia lamblia. American Journal of Hygiene. 5, 250-257. Hegner, RW. 1926 (a). Animal infections with the trophozoites of intestinal protozoa and their bearing on the functions of cysts. American Journal of Hygiene 6, 593-601. Hegner RW.1926(b). The biology of host-parasite relationships among protozoa living in man. Quartely Reviews of Biology. 1 , 393-418. Hegner R. 1927. Excystation in vitro of human intestinal protozoa. Science. 65, 577-578. Heinz M. 1988. Parasitology In Focus. Facts and Trends. Springler-Verlag; Berlin, 159
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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
Page 129 and 130: were evaluated. 5,2,1,2 Biosate Bio
Page 131 and 132: Table 5.1 Outlines a relative semi-
Page 133 and 134: 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: Farthing MJG, Mata L, Urrutia JJ, K
Page 183 and 184: patients with X-linked agammaglobul
Page 185 and 186: Press, Calgary. pp 57-58. Nash TE,
Page 187 and 188: of Entamoeba histolytica in a group
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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