Lead Toxicity in Mute Swans

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MATERIALS AND METHODS Collection of samples Blood samples (5ml) were collected as part of a major study on lead poisoning in Mute swans at Cork Lough, Co.Cork, SW Ireland (see O'Halloran et al., in press). Manually restrained (unanaesthetised) birds were blood sampled from the brachial vein, or in the case of cygnets from the tarsal vein, using a 21 gauge 1 inch needle. All samples were taken each day between 10.00-11.30. Blood was placed in lithium heparin tubes. An aliquot was also placed in tubes containing dipotassium salt of ethylene diamine tetra-acetic acid (EDTA). For serial sampling, four live birds were caged for 15 hours (16.00-07.00) in pens, 4m X lm, for the blood sampling procedure and subsequently released. Food and water were freely available at all time. Sex was determined by cloacal examination (after Hochbaum, 1942) and age was determined by plumage characteristics and beak colour. birds with brown feathers were designated as immatures. Cygnets and any Details of moulting status were noted. In six lead poisoned birds, four Mute and two Whooper swans Cygnus cygnus (L), blood samples were obtained prior to death. Tissue samples (liver, kidney, pancreas, breast muscle, heart muscle and gizzard muscle) were taken at post-mortem from lead poisoned swans for lead analyses. Blood chemistry and tissue analyses. Haemoglobin (Hb) content was determined by converting all haem species to ·alkaline haematin by techniques described elsewhere (see O'Halloran, et al., 1987b). Haematocrit was measured following - 89 -
I centrifugation in microhaematocrit tubes for · 30 mins at 12,000 x G (determined stroboscopically) in a Hawksley microcentrifuge. Other blood chemical parameters were estimated as follows: Aliquots of blood specimens collected in lithium heparin tubes were spun in a microcentrifuge (10,000 X G) for 5 mins. The plasma was removed and placed in cuvettes for analysis. Analysis was carried out in a selective multichannel autoanalyser (Hitachi Model, 737). A scanning wavelength of 340 to 700 nm was set, using a Halogen lamp source and a 0.6 cm light path. Calibration was based on Serva validated reference serum and normal saline (0.9% W/V), was used as a blank. Protoporphyrin ( free red blood cell protoporphyrin) was determined following a double phase extraction technique (after Peter~ al., 1978). Protoporphyrin IX standards were obtained from Porphyrin Products (Logan, Utah, U.S.). Fluorescence was measured on a Farrand Mark 1 Spectrofluoremeter with a primary filter of 405nm and a secondary filter of 600nm using a Xenon lamp and lOnm slit widths. Lead and protoporphyrin values were related to haemoglobin content (after O'Halloran et al., in press). Lead levels in blood and tissues were estimated following acid digestion, using an SP192 Flameless Atomic ~bsorption Spectrophotometer (for details see O'Halloran and Duggan, 1984). Liver and kidney lead levels in excess of 12.50 ug/g and 31.25 ug/g wet matter were considered diagnostic of lead poisoning after Clarke and Clarke (1975). Statistics. Blood chemistry and haematological reference distributions were calculated using a reference value statistical package based on the - 90 -
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0004067t:.7 1~11111 /I llll II llll
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This thesis is dedicated to my moth
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ABSTRACT Lead toxicity in Mute swan
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- what are the best methods for the
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Benson, W.W., Brock, O.W., Gabrica,
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CHAPTER 1 DETERMINATION OF HAEMOGLO
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MATERIALS AND METHODS Blood was obt
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standards are now available, it is
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I ' ABSTRACT The use of blood lead
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developed and (c) information on bl
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MATERIALS AND METHODS Study sites.
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Swans were sexed by cloacal examina
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Red blood cell protoporphyrin .. Th
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RESULTS Blood Lead Levels Diel vari
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16.00 24.CO 2.00 7.00 - ~ ~--1_ J l
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for corrected lead values (ug /g Rb
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Table 2. Blood lead levels of indiv
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30 20 (A) n = 357 10 0-.99 1- 2- 3-
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Blood lead is very labile (Chamberl
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median value of lead in April is si
Page 43 and 44: elevated lead levels. (2) Haemoglob
Page 45 and 46: Chisolm, J.J. & Browne, D.R. (1975)
Page 47 and 48: Ohi, G., Seki, H., Akiyama, K. & Ya
Page 49 and 50: CHAPTER 3: SECTION 1 LEAD POISONING
Page 51 and 52: mammals' (Finley et al. 1976). This
Page 53 and 54: commute from areas up to 20km away
Page 55 and 56: TABLE 1: Causes of death in Irish s
Page 57 and 58: I 4. Coiled shot: malleable stainle
Page 59 and 60: CHAPTER 3: SECTION 2 LEAD POISONING
Page 61 and 62: I INTRODUCTION The importance of le
Page 63 and 64: 42,000 anglers visit the State each
Page 65 and 66: MATERIALS AND METHODS Study sites B
Page 67 and 68: 30KM f Fig.l Map of Ireland indicat
Page 69 and 70: lead. X-ray. They were subsequently
Page 71 and 72: RESULTS Blood levels Blood lead lev
Page 73 and 74: I Table.II Proportion of lead poiso
Page 75 and 76: I of the gizzard and oesophagus and
Page 77 and 78: Table IV. 11 (of 41) X-rayed swans
Page 79 and 80: Table V. Median water lead levels (
Page 81 and 82: of pellets had occured. What is sig
Page 83 and 84: irds by Bellrose (1959) in the U.S.
Page 85 and 86: I the pellets as they moved in shor
Page 87 and 88: i Chamberlain, A.G. (1985). Predict
Page 89 and 90: Ong, C.N. & Lee, W.R. (1980). Inter
Page 91 and 92: SUMMARY Reference levels of some bi
Page 93: I adults, immatures, females and ma
Page 97 and 98: RESULTS All blood samples included
Page 99 and 100: Table 1. Mean (± s.e.) maximum and
Page 101 and 102: Table 3. Haematological and biochem
Page 103 and 104: Table 5. Haemoglobin, haematocrit a
Page 105 and 106: Table 6. Age, sex, blood lead level
Page 107 and 108: DISCUSSION The normal range of haem
Page 109 and 110: considered when using protoporphyri
Page 111 and 112: I the blood constituents, reflectin
Page 113 and 114: REFERENCES Bates, F.Y., Barnes, D.M
Page 115 and 116: i Lucas, A.M. and Jamroz, C. (1961)
Page 117 and 118: CHAPTER 5 TISSUE LEAD LEVELS AND HA
Page 119 and 120: INTRODUCTION Lead poisoning through
Page 121 and 122: I r MATERIALS AND METHODS Collectio
Page 123 and 124: I RESULTS Tissue lead levels Tissue
Page 125 and 126: Table 1. Tissue lead levels (ug/ g)
Page 127 and 128: Table 3 Age, sex, blood lead level
Page 129 and 130: DISCUSSION Tissue lead levels have
Page 131 and 132: which had collided had raised blood
Page 133 and 134: REFERENCES Anders, E., Dietz, D.D.,
Page 135 and 136: O'Halloran, J., Myers, A.A. and Dug
Page 137 and 138: many others for helping to catch sw
Page 139 and 140: APPENDIX 1 LEAD LEVELS IN MUTE SWAN
Page 141 and 142: intense fishing in summer and large
Page 143 and 144: area of 4m 2 , were taken. The samp
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levels of the birds on the Lee also
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i oesophagus, proventriculus and gi
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in blood. In general, healthy birds
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toxic lea? l~vels. The post-mortem
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London area. Part 2 Biological Effe
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Prelin1inary results of ringing Mut
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Ringed Mute S1rans ?f the. 61 I r~~
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Acknowledgments Ringed Mute S1ra11s
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Lead Toxicity in Mute Swans

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