in vitro culture and isoenzyme analysis of giardia lamblia

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Fig. 1.2. Life-cycle of Giardia lamblia. Ingestion of cysts results in excystation in the duodenum and subsequent colonisation of upper small intestine. 9
1.3 Transmission 1.3.1 Human hosts The cyst is spread through the faecal-oral route. Different mechanisms of transmission have been documented such as direct person to person hand-mouth transfer. This is more common in children (Isaac-Renton et al., 1993) owing to lack of personal hygiene and faecal incontinence. Prevalence is higher in children than in adults in endemic areas (Adam, 1991; Flanagan, 1992). There is suggestive evidence that institutionalised persons and day care attendees are at higher risk of infection, for example Cody and colleagues (1994) isolated cysts from chairs and tables in day care centres. In such settings, transmission rates are expectedly very high, and can subsequently enhance the overall transmission of the parasite. To illustrate this, day care attenders were shown to spread Giardia infection to the community in a controlled study by Polis and co-workers (1986). In this study, data suggested that 47% of the children in the centre transmitted giardiasis to at least one household contact. Although humans of all ages are susceptible to infection with Giardia, the prevalence is higher in children (Farthing et al., 1986) and infection frequently occurs in immunocompromised individuals (Webster, 1980). Inadequate sanitation exacerbates dissemination of cysts to the environment. Surface water supplies are also known to be contaminated with cysts and infected humans replenish this reservoir. Furthermore, wild and domestic animals such as beavers, muskrats, dogs and cats have also been implicated in transmission but their potential as zoonotic reservoirs of giardiasis is still debatable (Erlandsen, 1994). Prolonged occurrence of viable cysts in fresh cool (4°C) water has resulted in Giardia being reported as the most common aetiologic agent of epidemic 10
Page 1 and 2: IN VITRO CULTURE AND ISOENZYME ANAL
Page 3 and 4: (j)edicated to my {ate parents :No
Page 5 and 6: the positive specimens by sucrose g
Page 7 and 8: Declaration This study represents o
Page 9 and 10: ACKNOWLEDGEMENTS The author wishes
Page 11 and 12: Table of Contents Page CHAPTER 1 -
Page 13 and 14: 4.2.1. Animal Sourcing and Care ...
Page 15 and 16: APPENDIX 8 List of enzyme systems u
Page 17 and 18: List of Tables Page Table 1.1 Commo
Page 19 and 20: List of Plates Page Plate 2.1 An eo
Page 21 and 22: ABBREVIATIONS AND UNITS OF MEASUREM
Page 23 and 24: agi/is in 1882. In 1914, Alexeieff
Page 25 and 26: unique about Giardia in humans, whi
Page 27 and 28: To-date, use of the different speci
Page 29: Differentiation into the cyst stage
Page 33 and 34: Owen, 1984) 1.3. 2 The Zoonosis of
Page 35 and 36: 1.4 Distribution The morbidity and
Page 37 and 38: children by Schutte et al. in 1981
Page 39 and 40: of (a) the enormous absorptive capa
Page 41 and 42: Ig M (100% of patients), IgG (70%),
Page 43 and 44: chymotrypsin) between different iso
Page 45 and 46: impact on the physical growth of 29
Page 47 and 48: Figure 1.3. Transmission Electron m
Page 49 and 50: among children between the ages of
Page 51 and 52: where immunocompromised mice have p
Page 53 and 54: up to 4 distinct genotypes within e
Page 55 and 56: in initiation of infection (excysta
Page 57 and 58: In order to address these and many
Page 59 and 60: multiple stool examinations in 4 of
Page 61 and 62: 2.2 MATERIALS AND METHODS 2.2.1 Spe
Page 63 and 64: Table 2.1. Semi-quantitative assess
Page 65 and 66: 2.2.3.2 The modified Zinc sulphate
Page 67 and 68: Preliminary work with five samples
Page 69 and 70: % Viability = unstained cysts/total
Page 71 and 72: Table 2.3. Prevalence of Giardia in
Page 73 and 74: Table 2. 4. Monthly summary of stoo
Page 75 and 76: spp (59%). Other parasites less fre
Page 77 and 78: Observations • A large proportion
Page 79 and 80: Plate 2.1. An eosin-stained prepara
Page 81 and 82:
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
Page 103 and 104:
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
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Table 5.2. Sensitivity to Ciproflox
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Table 5.3. Summary results of in vi
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Once established in culture for a l
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Plates 5.3 and 5.4 Confluent growth
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difficult to produce viable culture
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dominance of certain genotypes unde
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Cryopreservation and successful ret
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adjusted to 5°C per minute from am
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6.3 RESULTS Trophozoites have been
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Table 6.1. A longevity record of sa
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Diamond (1995) stated that an impor
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into two categories. One group cont
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technique to the first local (South
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for Giardia Iysates. These included
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4. To exclude the presence of bacte
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Plate 7.1 Banding pattern of 8 seri
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PGM activity was displayed by Giard
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Plate 7.6. Hexokinase Lane 1. Unino
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Plate 7.8. Phosphogluconate dehydro
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stably expressed between different
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techniques, which would allow diffe
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Belosevic M, Faubert GM, MacLean JD
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Farthing MJG, Mata L, Urrutia JJ, K
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Heidelberg pp.397-398. Hiatt RA, Ma
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patients with X-linked agammaglobul
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Press, Calgary. pp 57-58. Nash TE,
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of Entamoeba histolytica in a group
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Wolfe M. S.1979. Giardiasis. Sympos
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APPENDICES APPENDIX 1 Information t
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Unayo imvume yokunqaba ukungenela l
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Appendix 3 Excystation and culture
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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
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Appendix 10. Quantitative descripti
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in vitro culture and isoenzyme analysis of giardia lamblia

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