Acid-fast Intranuclear Inclusion Bodies Research Articles

Lead Poisoning in Whooper and Tundra Swans

Six weak whooper swans (Cygnus cygnus) and two weak tundra swans (Cygnus columbianus) were found at Swamp Miyajima (Hokkaido, Japan) in May 1998. Anorexia, depression, green watery feces, pale conjunctiva, and anemia were observed. Radiographs showed from six to 38 suspected lead pellets in the gizzard. Blood lead concentrations were 2.5-6.7 microg/g (mean+/-SD=4.6+/-1.14 microg/g) on day 1. After blood collection, the birds were treated with calcium disodium ethylenediaminetetraacetate (CaEDTA) given intravenously and force fed. Despite treatment, seven birds died the next day. Green, bile-stained livers and pale or green kidneys were observed on necropsy. Microscopically, bile pigment was widespread in the liver and acid-fast intranuclear inclusion bodies were observed in renal tubular epithelium. Lead concentrations in livers and kidneys were 14.0-30.4 microg/g and 30.2-122 microg/g wet weight, respectively. Only one bird survived and this whooper swan continued to be treated with CaEDTA and activated charcoal. No lead shot was observed in the proventriculus and gizzard by radiography on day 64 and the blood lead concentration decreased from 2.9 microg/g to 0.09 microg/g during that same period. After 4 mo of rehabilitation, the whooper swan was returned to the wild. Lead intoxication continues to be a problem at Swamp Miyajima.

Toxicity of lead-contaminated sediment to mute swans.

Most ecotoxicological risk assessments of wildlife emphasize contaminant exposure through ingestion of food and water. However, the role of incidental ingestion of sediment-bound contaminants has not been adequately appreciated in these assessments. This study evaluates the toxicological consequences of contamination of sediments with metals from hard-rock mining and smelting activities. Lead-contaminated sediments collected from the Coeur d'Alene River Basin in Idaho were combined with either a commercial avian maintenance diet or ground rice and fed to captive mute swans (Cygnus olor) for 6 weeks. Experimental treatments consisted of maintenance or rice diets containing 0, 12 (no rice group), or 24% highly contaminated (3,950 microg/g lead) sediment or 24% reference (9.7 microg/g lead) sediment. Although none of the swans died, the group fed a rice diet containing 24% lead-contaminated sediment were the most severely affected, experiencing a 24% decrease in mean body weight, including three birds that became emaciated. All birds in this treatment group had nephrosis; abnormally dark, viscous bile; and significant (p <or= 0.05) reductions in hematocrit and hemoglobin concentrations compared to their pretreatment levels. This group also had the greatest mean concentrations of lead in blood (3.2 microg/g), brain (2.2 microg/g), and liver (8.5 microg/g). These birds had significant (alpha = 0.05) increases in mean plasma alanine aminotransferase activity, cholesterol, and uric acid concentrations and decreased plasma triglyceride concentrations compared to all other treatment groups. After 14 days of exposure, mean protoporphyrin concentrations increased substantially, and mean delta-aminolevulinic acid dehydratase activity decreased by more than 95% in all groups fed diets containing highly contaminated sediments. All swans fed diets that contained 24% lead-contaminated sediment had renal acid-fast intranuclear inclusion bodies, which are diagnostic of lead poisoning in waterfowl. Body weight and hematocrit and hemoglobin concentrations in swans on control (no sediment) and reference (uncontaminated) sediment diets remained unchanged. These data provide evidence that mute swans consuming environmentally relevant concentrations of Coeur d'Alene River Basin sediment developed severe sublethal lead poisoning. Furthermore, toxic effects were more pronounced when the birds were fed lead-contaminated sediment combined with rice, which closely resembles the diet of swans in the wild.

Pathological Study of Lead Poisoning in Whooper Swans (Cygnus cygnus) in Japan

During spring 1989, thirty-three whooper swans (Cygnus cygnus) died at Lake Miyajima in Hokkaido, Japan; 15 were examined. The birds were diagnosed as having subacute lead poisoning due to ingestion of spent lead shot. The main gross findings were bile-stained liver, edematous or gelatinous bone marrow, bile-stained lining with hyperkeratosis and lead pellets in the gizzard, and proventricular impaction. Histopathologically, there was lead-hemolytic jaundice of the liver, hemosiderosis in the liver and spleen, and hypoplasia of the bone marrow with increased numbers of polychromatic erythroblasts. Acid-fast intranuclear inclusion bodies were seen in kidneys of seven swans. Under electron microscopy, inclusion bodies had frayed contours and consisted of high-electron-dense fine granules. The lead concentration of the liver ranged from 5.5 to 44.3 mg/kg wet weight. It was suggested that these changes resulted from excess breakdown of erythrocytes, inhibition of heme synthesis, and impaired erythropoiesis caused by lead shot.

Detection and cellular localization of lead by electron probe analysis in the diagnosis of suspected lead poisoning in rhesus monkeys.

Lead poisoning of unknown source was diagnosed histologically in 2 rhesus monkeys (Macaca mulatta) by finding acid-fast intranuclear inclusion bodies in the epithelial cells of renal cortical tubules. The presence of lead in the inclusions was determined by scanning electron microscopy/energy dispersive x-ray analysis using sections from paraffin embedded tissues. This observation indicates the usefulness of this technique for the detection and cellular localization of lead in tissues, even from archival material.

Paint chip poisoning of Laysan albatross at Midway Atoll.

Epizootic mortality occurred in Laysan albatross (Diomedea immutabilis) fledgings at Midway Atoll in 1983. Heavy metal toxicity from ingestion of weathered paint chips was one of the causes. Sick albatrosses were unable to retract their wings, causing a "droop-wing" appearance. Five normal and 12 droop-winged fledglings were captured, killed, and examined. Paint chips found in the proventriculus of the affected fledglings contained up to 144,000 ppm lead. Blood, liver, and kidney concentrations of lead in affected birds were higher than in normal fledglings, and acid-fast intranuclear inclusion bodies were present in the kidneys. Degenerative lesions were present in the myelin of some brachial nerves. Weathered paint samples collected from 12 buildings contained up to 247,250 ppm lead and 101 ppm mercury. Lead poisoning was diagnosed in 10 of the droop-winged albatrosses and was one of the causes of morbidity. Mercury toxicosis and plastic impaction were other possible causes.

Metal contamination in wildlife living near two zinc smelters

Wildlife in an oak forest on Blue Mountain was studied 10 km upwind (Bake Oven Knob site) and 2 km downwind (Palmerton site) of two zinc smelters in eastern Pennsylvania, USA. Previous studies at sites near these smelters had shown changes in populations of soil microflora, lichens, green plants and litter-inhabiting arthropods. The 02 soil litter horizon at Palmerton was heavily contaminated with Pb (2700 mg kg −1), Zn (24 000 mg kg −1), and Cd (710 mg kg −1), and to a lesser extent with Cu (440 mg kg −1). Various kinds of invertebrates (earthworms, slugs and millipedes) that feed on soil litter or soil organic matter were rare at, or absent from, the Palmerton site. Those collected at Bake Oven Knob tended to have much higher concentrations of metals than did other invertebrates. Frogs, toads and salamanders were very rare at, or absent from, the Palmerton site, but were present at Bake Oven Knob and at other sites on Blue Mountain farther from the smelters. Metal concentrations (dry wt) in different organisms from Palmerton were compared. Concentrations of Pb were highest in shrews (110 mg kg −1), followed by songbirds (56 mg kg −1), leaves (21 mg kg −1), mice (17 mg kg −1), carrion insects (14 mg kg −1), berries (4·0 mg kg −1), moths (4·3 mg kg −1) and fungi (3·7 mg kg −1). Concentrations of Cd, in contrast, were highest in carrion insects (25 mg kg −1), followed by fungi (9·8 mg kg −1), leaves (8·1 mg kg −1), shrews (7·3 mg kg −1), moths (4·9 mg kg −1), mice (2·6 mg kg −1), songbirds (2·5 mg kg −1) and berries (1·2 mg kg −1). Concentrations of Zn and Cu tended to be highest in the same organisms that had the highest concentrations of Cd. Only a small proportion of the metals in the soil became incorporated into plant foliage, and much of the metal contamination detected in the biota probably came from aerial deposition. The mice from both sites seemed to be healthy. Shrews had higher concentrations of metals than did mice, and one shrew showed evidence of Pb poisoning; its red blood cell ALAD activity was greatly reduced, one kidney contained acid-fast intranuclear inclusion bodies, and the other kidney had 280 mg kg −1 Pb (wet wt). The livers from two cuckoos from Palmerton had elevated Pb concentrations (18 and 25 mg kg −1, wet wt). Most of the songbirds seemed to be healthy.

A Comparative Study of Lead Nephrotoxicity and Tissue Deposition Between Rats, Mice, and Pigeons Exposed to Drinking Water Containing 2, 20, or 200 ppm Lead

White Carnaew pigeons were exposed to drinking water containing various concentrations of lead to study the relationship between exposure and tissue deposition and the relationship between the concentrations of lead in kidney and morphologic alterations. The concentration of lead in the blood was the only tissue which showed a linear increase with respect to the concentration of lead in the drinking water. The concentration of lead in the liver, kidney, brain, heart, aorta, and pancreas increased in response to lead in the drinking water; however, these increases were not linear with respect to lead exposure. Similar results were obtained for rats and mice exposed via the drinking water to 2, 20, or 200 ppm lead; however, the concentration of lead in tissues was substantially greater in pigeons. Kidneys from rats and mice appeared normal following the exposure of 200 ppm lead for 4 months, whereas, in kidneys from pigeons exposed to 200 ppm for a similar period of time there was tubular degeneration, fibrosis, and acid fast intranuclear inclusion bodies. These pathoanatomic alterations may be related to the concentration lead found in the kidney in pigeons. The concentration of lead in the kidney was 3,930 μg gram dry weight for pigeons but only 40 and 6 μg for mice and rats respectively. These studies suggest that the pigeon is more responsive to ingested lead than the rat or mouse. The renal lesions induced by chronic lead exposure are similar in pigeons and man suggesting that the pigeon may be useful animal model to study the yet unanswered questions related to chronic nephritis following lead poisoning.

Lead poisoning in captive wild animals.

Lead poisoning was diagnosed post-mortem in 34 simian primates, 11 parrots, and 3 Australian fruit bats at the National Zoological Park. Diagnoses were made by the finding of acid-fast intranuclear inclusion bodies in renal epithelia or hepatocytes and, in most cases, by finding excess lead in samples of liver. The estimated prevalence of lead intoxication among autopsied primates and parrots was 44% and 50% respectively. Leaded paint was found in many animal enclosures at this zoo and it was available to all the lead-poisoned animals in this study. The finding of renal intranuclear inclusion bodies in animals at several zoos, scattered reports of lead intoxication of animals dwelling in various zoos, the occurrence of leaded paint in many zoos and the high incidence of lead poisoning at this zoo, indicated that lead poisoning of zoo animals is much more common than was previously thought.

Relative Toxicity of Lead and Five Proposed Substitute Shot Types to Pen-Reared Mallards

A 30-day toxicity test was made to determine the relative toxicity of lead, a tin-lead alloy, zinc, nickel, teflon-coated steel, and tin, all in shot form, to pen-reared mallard drakes. All of the 15 ducks dosed with lead died. Twenty-seven percent of 15 dosed with alloy, and 20 percent of 15 dosed with zinc also died. Ten of the remaining zinc-dosed ducks showed signs of distress, including losses of muscular control and body weight. There were no deaths among 15 ducks dosed with nickel, 15 dosed with teflon-coated steel, and 15 dosed with tin. Seventy-three percent of those dosed with nickel shot eliminated all shot before the end of the 30-day period. Acid-fast intranuclear inclusion bodies were present in the kidneys of mallards dosed with commercial lead shot, or with tin-lead alloy shot, but not in the kidneys of birds given nickel, tin, or teflon-coated steel shot. Atypical, pale, acid-fast bodies were found in kidneys of 1 of 15 birds dosed with zinc. An iron-containing pigment, which stained positive with the Prussian blue technique, was present in variable amounts in almost all livers. Zinc-dosed ducks that died or were killed while still showing signs of zinc intoxication had higher iron levels in the liver than ducks that had recovered from zinc intoxication. Mortality of waterfowl due to lead poisoning has been recognized for many years. Despite several research studies (Green and Dowdell 1936, Dowdell and Green 1937, and Jordan and Bellrose 1950) no satisfactory substitute for the presently used commercial lead shot has been developed. In 1965 the Patuxent Wildlife Research Center began a research project aimed at the development of a non-toxic shot. In 1966 the Sporting Arms and Ammunition Manufacturers' Institute and the Patuxent Wildlife Research Center undertook this study as a cooperative research project. Metals or alloys proposed as possible substitutes for lead are first tested for their acute toxicitx to pen-raised mallards. Candidate materials found to be non-toxic are then tested for possible effects on reproduction, for ballistical properties, and for killing power. The results of the first group of candidate materials tested were reported by Irby et al. 1967, and Locke et al. 1967. 1 Present address: Department of Forestry and Wildlife Management, University of Massachusetts, Amherst. This paper reports the results of the acute toxicity tests for another group of candidate materials, and is part of a series of studies designed to develop a non-toxic shotgun pellet. We acknowledge the assistance of John L. Sincock, Ralph Andrews, Larry T. Young, and Robert G. Heath, all of the Pa uxent Wildlife Research Center. MATERIALS AND METHODS The birds tested were 18-month-old mallard drakes (Anas platyrhynchos) reared at the Patuxent Wildlife Research Center. One hundred and five birds were included in the experiment. For 9 months prior to the study they were kept together in a single pen and fed a mixture of commercial duck pellets and mixed grains. During the study they were kept in groups of five in 21 pens 30 x 15 x 6 ft high and offered cracked corn, quartz, and oystershell grit ad libitum. Each pen contained a feeding trough and a watering trough at ground level, with flowing water continually maintained at a depth of 10 inches.

Histopathology of Mallards Dosed With Lead and Selected Substitute Shot

The histopathological response of male game farm mallards fed lead, three types of plastic-coated lead, two lead-magnesium alloys, iron, copper, zinc-coated iron, and molybdenum-coated iron shot was studied. Mallards fed lead, plastic-coated lead, or lead-magnesium alloy shot developed a similar pathological response, including the formation of acid-fast intranuclear inclusion bodies in the kidneys. Birds fed iron or molybdenum-coated iron shot developed hemosiderosis of the liver. Two of four mallards fed zinc-coated iron shot also developed hemosiderosis of the liver. No lesions were found in mallards fed copper shot.

Lead Poisoning in a Sample of Maryland Mourning Doves

A sick mourning dove (Zenaidura macroura) collected in Maryland with 2 lead shot in the gizzard showed acid-fast intranuclear inclusion bodies in the kidney tubular cells. The liver and the tibia contained 72 ppm and 187 ppm lead (wet weight) respectively. Four gizzards from 62 doves killed by hunters contained lead shot. The lead content of 43 dove livers ranged from 0.4-14.0 ppm (wet weight); 40 of these doves were collected by hunters, and the other 3 were dying of trichomoniasis.

Acid-Fast Intranuclear Inclusion Bodies in the Kidneys of Mallards Fed Lead Shot

Acid-fast intranuclear inclusion bodies were found in the cells of the proximal convoluted tubules of the kidneys of mallards fed one, two, three or eight number 6 lead shot and maintained on cracked or whole corn and on grain-duck pellet diets. No acid-fast inclusion bodies were found in mallards fed one or three lead shot but maintained on a duck pellet ration. Dietary factors may be responsible for the failure of mallards fed a duck pellet ration to develop lead inclusion bodies when treated with one or three lead shot. The authors suggest these inclusion bodies can be used as presumptive evidence for lead intoxication in mallards.