Reproductive effects and duckling survivability following chronic dosing with tungsten-iron and tungsten-polymer shot in adult game-farm mallards

Tungsten-iron and tungsten-polymer shot were given conditional approval for waterfowl hunting by the U.S. Fish and Wildlife Service based partly on the results of a 30-day acute toxicity trial utilizing mallards (Anas platyrhynchos). Final approval of the two tungsten-containing shot was contingent on the results of a 150-day study that assessed the health and reproductive effects of tungsten-iron and tungsten-polymer shot in adult mallards. Reproductive data are presented in this paper. Sixteen male and 16 female adult mallards were dosed orally with eight #4 steel shot (control), eight #4 tungsten-iron shot, or eight #4 tungsten-polymer shot on days 0, 30, 60, 90, and 120 of a 150-day trial (26 January 1998 to 25 June 1998). Reproductive performance was assessed during the last 90 days (day 61 to day 150) of the trial. There were no significant differences in egg production and fertility and hatchability of eggs from tungsten-iron- and tungsten-polymer-dosed ducks compared to control ducks. There was no evidence of differences in percent survivability and body weight of ducklings from tungsten-iron and tungsten-polymer mallards compared to ducklings from control ducks. Tungsten-iron or tungsten-polymer shot repeatedly administered to adult mallards during the 150 day trial did not adversely affect reproduction or their offspring.     

Health effects following chronic dosing with tungsten-iron and tungsten-polymer shot in adult game-farm mallards

Permanent approval of shot composed of tungsten-iron and tungsten-polymer for waterfowl hunting by the U.S. Fish and Wildlife Service was pending the results of the present study that examined the health and reproductive effects of the two shot types on mallards (Anas platyrhynchos) over a 150-day period. We collected data pertaining to the effects of tungsten-iron and tungsten-polymer shot on mortality, body weight, organ weight, tissue pathology, and shot erosion. Thirty-two bird groups (sexes equal) of adult mallards were dosed orally with eight #4 steel shot (control), eight #4 tungsten-iron shot, or eight #4 tungsten-polymer shot on days 0, 30, 60, 90, and 120 of a 150-day trial (26 January 1998 to 25 June 1998). An additional 12 mallards (sexes equal) were dosed orally with eight #4 lead shot (positive control) on day 0 of the study. All lead-dosed ducks died by day 25, whereas no ducks died in the other treatment groups. Significant liver hemosiderosis was present in all control and tungsten-iron-dosed males, in five of eight control and three of eight tungsten-iron-dosed females, and in one tungsten-polymer-dosed male examined. The rate of shot erosion was highest for tungsten-polymer shot (99%), followed by tungsten-iron (72%), and steel (55%) shot. Tungsten-iron or tungsten-polymer shot repeatedly administered to adult mallards did not have deleterious health effects during the 150-day trial based on mortality, body weights, organ weights, and histology of the liver and kidneys.     

Acute toxicity of lead, steel, and an iron-tungsten-nickel shot to mallard ducks (Anas platyrhynchos)

Twenty mallards (Anas platyrhynchos) of both sexes were dosed by oral gavage with Heavi-Shot (H-S; Environ-Metal, Inc., Sweet Home, Oregon, USA) pellets, 20 with steel shot, and 10 with lead (Pb) pellets, all of equal size. All pellets were fired from a shotgun into an absorbent material, retrieved, and weighed prior to introduction into the ducks. Birds were fed whole kernel corn and grit and observed for signs of toxicity for 30 days following dosing. Hevi-Shot pellets lost an average of 6.2% of their mass and steel shot pellets lost 57% of their mass in the birds' gizzards. Almost all (90%) of the Pb shot dosed birds died before the end of the study, while no mortality was observed in the steel or H-S dosed groups. Even though total food consumption differed between the H-S and steel shot groups, mean bird weight change was not different. There were no significant morphologic or histopathologic abnormalities of the liver and kidney in the H-S and steel shot groups. Results indicated that mallards dosed orally with eight No. 4 H-S pellets were not adversely affected over a 30-day period, and that H-S provides another environmentally safe nontoxic shot for use in waterfowl hunting.     

Influence of diet on the hematology and serum biochemistry of zinc-intoxicated mallards

Changes in hematological and serum biochemistry parameters in female zinc (Zn)-dosed farm-raised mallards (Anas platyrhynchos) fed four different diets were examined. Sixty ducks received an average dose of 0.97 g of Zn in the form of eight, 3.30-mm diameter shot pellets containing 98% Zn and 2% tin, and another 60 ducks were sham-dosed as controls. Fifteen ducks from each of the two dosing groups were assigned to one of four dietary treatments: corn only, corn with soil, commercial duck ration only, or commercial duck ration with soil. Shot-pellet dissolution rates ranged from 7 mg/Zn/day to 27 mg/Zn/day. Regardless of diet, the Zn dose resulted in mortality; incoordination; paralysis and anorexia; decreased body, liver, pancreas, gonad, and gizzard weight; increased kidney weight; and macroscopic lesions. Zn-dosed ducks had a lower mean erythrocyte packed cell volume (PCV), higher mean reticulocyte count, and a greater number of individuals with immature and/or abnormal erythrocytes, than did control mallards. Mean total leucocyte counts were higher in Zn-dosed ducks than in controls. Zn-dosed ducks that had soil available had higher leucocyte counts than those without soil. Zn-dosed ducks were characterized by a marked heterophilia and relative lymphopenia. In Zn-dosed ducks, the mean lymphocyte count was highest in those provided a commercial duck ration, and lowest in those fed corn. In control ducks, the mean lymphocyte count was highest in ducks fed corn, and lowest in those provided soil along with a commercial duck ration. Zn-dosed mallards had higher serum aspartate aminotransferase and amylase levels, and lower alkaline phosphatase activities than control ducks. Serum phosphorus and uric acid concentrations were higher, and calcium, glucose, and total protein levels lower, in Zn-dosed ducks than in control ducks. Diet did affect serum calcium, phosphorus, total protein, and uric acid concentrations. Differences in erythrocyte and leucocyte parameters, serum enzyme activities, and metabolite concentrations were associated with dose and diet effects. Diets high in protein and other organic matter and calcium and phosphorus did not prevent or substantially alleviate Zn toxicosis in farm-raised mallard ducks.     

Comparative toxicity of lead shot in black ducks (Anas rubripes) and mallards (Anas platyrhynchos)

In winter, pen-reared and wild black ducks (Anas rubripes), and game farm and wild mallards (Anas platyrhynchos), maintained on pelleted feed, were sham-dosed or given one number 4 lead shot. After 14 days, dosed birds were redosed with two or four additional lead shot. This dosing regimen also was repeated in summer using pen-reared black ducks and game farm mallards. Based upon mortality, overt intoxication, weight change, delta-aminolevulinic acid dehydratase activity and protoporphyrin concentration, black ducks and mallards were found to be equally tolerant to lead shot. However, captive wild ducks were more sensitive than their domesticated counterparts, as evidenced by greater mortality and weight loss following lead shot administration. This difference may be related to stress associated with captivity and unnatural diet.     

Effects of ingested lead on antibody production in mallards (Anas platyrhynchos)

We examined the effects of lead ingestion on in vitro and in vivo indices of immune function in mallards (Anas platyrhynchos). Twenty-four mallard drakes were randomly divided into three groups (I, II, III), then assigned to treatment or control subgroups (n = 4). On day 0, all treatment birds were dosed orally with one number 4 lead shot (mean = 0.2 g). We challenged all individuals in each group with washed sheep red blood cells (SRBC) injected intraperitoneally on days 0 (Group I), 7 (Group II) or 14 (Group III), and collected blood for analyses 7 and 8 days after SRBC challenge. We measured and compared blood lead concentrations, in vitro lymphocyte transformation responses to phytohemagglutinin A and lipopolysaccharide, and hemagglutination titers to SRBC. Mean blood lead concentrations were elevated (P less than or equal to 0.04) in treatment birds at each sampling period. Large individual variability in lymphocyte stimulation responses precluded further analysis of those data. Hemagglutination titers to SRBC were lower (P less than 0.0001) in lead-poisoned ducks than in controls, suggesting that ingested lead may have immunosuppressive effects on mallards.     

A comparison of two methods to establish the prevalence of lead shot ingestion in mallards (Anas platyrhynchos) from The Netherlands

Two collection methods for screening the mallard (Anas platyrhynchos) population in the Netherlands for the ingestion of spent lead shot were compared. One method consisted of examination of gizzards from mallards shot by hunters (n = 2,859) and the other method consisted of examination of gizzards from mallards caught in duck traps (n = 865). The 95% confidence interval of lead shot ingestion in the mallard population estimated by the first method was 1.7 to 2.9% and by the second method 1.1 to 3.1%. These values were not significantly different. From the numbers of lead pellets embedded in the gizzard wall in hunter-killed and trapped mallards it was estimated that at least 22 to 68% of the trapped ducks had been hit by lead shot previously, but survived. Furthermore, this study shows that it is reasonable to assume that a substantial part of the pellets which are identified (in this study and other studies) as ingested, may well have been shot into the gizzard lumen at some time before the birds were actually killed. To avoid lead poisoning in mallards and in raptors depredating waterfowl hit by lead shot, a change to steel shot is advocated.     

Genetic structure of Eurasian and North American mallard ducks based on mtDNA data

To elucidate the origin and genetic structure of the domesticated duck in Eurasia and North America, we sequenced 114 duck D-loop sequences and retrieved 489 D-loop sequences from GenBank. In total, 603 ducks including 50 duck breeds/populations from eight countries (China, France, Russia, India, Kazakhstan, Mongolia, Thailand and USA) were used in this study. One hundred and thirty-four haplotypes and 81 variable sites were detected. H49 was the predominant haplotype, which was considered to be the same dominant haplotype found in the previous studies, and was found in 309 birds. The smallest values for both genetic differentiation index (F(ST), 0.04156) and the number of the net nucleotide substitutions between two populations (D(A), 0.00018) were observed between Eurasian domestic ducks and Eurasian mallards. No geography, breed or population clusters were observed in the Eurasian domestic ducks and mallards. Five haplotypes were shared by USA mallards and Eurasian domestic duck/Eurasian mallards. Only one haplotype (H49) was shared by Eurasian domestic ducks and China spot-billed ducks. By combining phylogenetic analyses, haplotype network profile, genetic distances and shared haplotypes, we can draw two major conclusions: (i) Eurasian and North American mallards show a clear geographic distribution pattern; (ii) Eurasian domestic ducks are derived from the Eurasian mallards, not from the spot-billed ducks.     

Experimental inoculation of mallard ducks (Anas platyrhynchos platyrhynchos) with Borrelia burgdorferi

Birds have been incriminated as disseminaters of Borrelia burgdorferi and have the potential to spread the organism over a wide geographic range. Borrelia burgdorferi has been isolated from the liver and blood of passerine birds and from Ixodes dammini removed from passerines. The objective of this study was to determine if waterfowl, specifically mallards (Anas platyrhynchos platyrhynchos), were susceptible to infection with B. burgdorferi. Eight ducks were inoculated with B. burgdorferi; four orally and four intravenously (i.v.) and two ducks were inoculated with phosphate buffered saline as controls. All eight inoculated birds became infected and developed antibodies to B. burgdorferi. The spirochete was isolated from cloacal material from an orally infected duck on day 22 postinoculation (PI) and from an i.v. infected bird on day 29 PI, from the blood of an i.v. infected bird on day 7 PI, and from the kidney of an orally infected bird. Borrelia burgdorferi was detected by indirect immunofluorescence using the B. burgdorferi specific monoclonal antibody H5332 in kidneys of three orally infected birds and one i.v. infected bird and from the mesentery of one orally infected bird. These findings show that mallard ducks are susceptible to infection by B. burgdorferi and that they can be infected orally and shed the organism in the droppings. Thus, mallards could disseminate B. burgdorferi over long distances without the need of an arthropod vector.     

A Comparison of Hybridization between Mottled Ducks (<Emphasis Type="Italic">Anas fulvigula</Emphasis>)and Mallards (<Emphasis Type="Italic">A. platyrhynchos</Emphasis>) in Florida and South Carolina using Microsatellite DNA Analysis

Interspecific hybridization has been implicated in population declines for some waterfowl species within the mallard complex, and hybridization with mallards (Anas platyrhynchos) is currently considered the largest threat to mottled ducks (A. fulvigula), one North American member of that complex. We assessed genetic variation among 225 mottled ducks and mallards using five microsatellite loci, and detected significant overall differences between these species within two geographic areas. We characterized hybridization in Florida, where mottled ducks are endemic and mallards are beginning to appear on the breeding grounds, and in South Carolina, where mottled ducks were introduced outside their native range. We used Bayesian genetic mixture analysis in an attempt to distinguish between these closely related species. In Florida, we detected two distinct genetic groups, and 10.9% of our samples from Florida mottled ducks were inferred to have been hybrids. In contrast only 3.4% of Florida mallards were inferred to have been hybrids, suggesting asymmetric hybridization. Populations from different geographic areas within Florida exhibited hybridization rates ranging from 0% to 24%. These data indicate a genetic component would be appropriate in actively managing interspecific hybridization in Florida mottled ducks. In contrast, South Carolina mottled ducks and mallards cannot be differentiated.     

Influenza Virus in a Natural Host,the Mallard: Experimental Infection Data

Wild waterfowl, particularly dabbling ducks such as mallards (Anas platyrhynchos), are considered the main reservoir of low-pathogenic avian influenza viruses (LPAIVs). They carry viruses that may evolve and become highly pathogenic for poultry or zoonotic. Understanding the ecology of LPAIVs in these natural hosts is therefore essential. We assessed the clinical response, viral shedding and antibody production of juvenile mallards after intra-esophageal inoculation of two LPAIV subtypes previously isolated from wild congeners. Six ducks, equipped with data loggers that continually monitored body temperature, heart rate and activity, were successively inoculated with an H7N7 LPAI isolate (day 0), the same H7N7 isolate again (day 21) and an H5N2 LPAI isolate (day 35). After the first H7N7 inoculation, the ducks remained alert with no modification of heart rate or activity. However, body temperature transiently increased in four individuals, suggesting that LPAIV strains may have minor clinical effects on their natural hosts. The excretion patterns observed after both re-inoculations differed strongly from those observed after the primary H7N7 inoculation, suggesting that not only homosubtypic but also heterosubtypic immunity exist. Our study suggests that LPAI infection has minor clinically measurable effects on mallards and that mallard ducks are able to mount immunological responses protective against heterologous infections. Because the transmission dynamics of LPAIVs in wild populations is greatly influenced by individual susceptibility and herd immunity, these findings are of high importance. Our study also shows the relevance of using telemetry to monitor disease in animals.     

Sex reversal and aromatase in chicken.

Aromatase inhibitors administered before sexual differentiation of the gonads can induce sex reversal in female chickens. To analyze the process of sex reversal, we have followed for several months the changes induced by Fadrozole, a nonsteroidal aromatase inhibitor, in gonadal aromatase activity and in morphology and structure of the female genital system. Fadrozole was injected into eggs on day four of incubation, and its effects were examined during the embryonic development and for eight months after hatching. In control females, aromatase activity in the right and the left gonad was high in the middle third of embryonic development, and then decreased up to hatching. After hatching, aromatase activity increased in the left ovary, in particular during folliculogenesis, whereas in the right regressing gonad, it continued to decrease to reach testicular levels at one month. In treated females, masculinization of the genital system was characterized by the maintenance of the right gonad and its differentiation into a testis, and by the differentiation of the left gonad into an ovotestis or a testis; however, in all individuals, the left Müllerian duct and the posterior part of the right Müllerian duct were maintained. In testes and ovotestes, aromatase activity was lower than in gonads of control females (except in the right gonad as of one month after hatching) but remained higher than in testes of control and treated males. Moreover, in ovotestes, aromatase activity was higher in parts displaying follicles than in parts devoid of follicles. The main structural changes in the gonads during sex reversal were partial (in ovotestes) or complete (in testes) degeneration of the cortex in the left gonad, and formation of an albuginea and differentiation of testicular cords/tubes in the two gonads. Testicular cords/tubes transdifferentiated from ovarian medullary cords and lacunae whose epithelium thickened and became Sertolian. Transdifferentiation occurred all along embryonic and postnatal development; thus, new testicular cords/tubes were continuously formed while others degenerated. The sex reversed gonads were also characterized by an abundant fibrous interstitial tissue and abnormal medullary condensations of lymphoid-like cells; in the persisting testicular cords/tubes, spermatogenesis was delayed and impaired. Related to aromatase activity, persistence of too high levels of estrogens can explain the presence of oviducts, gonadal abnormalities and infertility in sex reversed females.     

Cytochrome P-450 and parathion metabolism in the fetal and adult gonads of the horse.

Mixed function oxidase-catalyzed dearylation of parathion was determined in microsomal fractions of gonadal and liver tissues of fetal and adult horse. When parathion metabolism was calculated on a basis of milligram of protein, the activity was found to be similar between fetal and adult gonads but not liver. However, on a basis of nmoles of cytochrome P-450, parathion metabolism was similar between fetal and adult liver but not gonadal tissues; the metabolism being 20 times higher in the adult than the fetal gonad.     

Mottled ducks,feral mallards,and their hybrids in Florida

The nonmigratory and endemic Florida mottled duck (Anas fulvigula fulvigula) is facing conservation threats from the combined effects of urbanization and introgressive hybridization with feral mallards (Anas platyrhynchos) and mallard x mottled duck hybrids. In the past, the status of the Florida mottled duck population was assessed during annual aerial surveys and most brown ducks (mottled ducks, mallards, and hybrids of them) detected during the survey would have been mottled ducks. But the release of domesticated mallards for aesthetic purposes has led to increases in the prevalence of mallards-hybrids (mallards or mallard x mottled duck hybrids) throughout peninsular Florida, USA, and because it is impossible to differentiate among mottled ducks, female mallards, and hybrids during aerial surveys, helicopter surveys were halted in 2009 until state researchers could conduct a range-wide study to determine what proportion of brown ducks are mottled ducks versus mallards-hybrids. We used plumage keys and high-resolution photography to categorize brown ducks from 557 wetland grid points as either mottled ducks or mallards-hybrids. Of the 5,179 brown ducks categorized, 40.1% were mottled ducks and 59.9% were mallards-hybrids. We used logistic regression analysis to model the interactive effect of a site's latitude and level of urbanization (urban gradient value within a 2-km buffer) to generate a predictive raster surface (1-km resolution) of the study area with values corresponding to the probability that a brown duck observed within a cell is a pure mottled duck. Predicted values will be used as correction factors when estimating final mottled duck population abundance from brown-duck survey data. Additionally, the predictive raster surface will be used to identify wetlands where mottled ducks remain predominant so that these sites can be targeted for preservation. Overall, mallards-hybrids outnumbered mottled ducks throughout most of peninsular Florida, especially in more urbanized regions, and their current prevalence rate presents a serious conservation threat, via hybridization, to extant mottled duck populations.     

Metal (Fe, Zn, Mn and Cu) levels in the Antarctic fish Notothenia coriiceps

During the 1994/1995 Antarctic summer 20 Antarctic cod (Notothenia coriiceps) of both sexes were collected at King George Island, South Shetland Islands, Antarctica, to determine the levels of Fe, Zn, Mn and Cu in muscle, liver, gonad and kidney. Metal levels were assessed using the inductively coupled plasma-atomic emission spectrometry (ICP-AES) technique. The levels of Fe, Zn, Mn and Cu observed in both sexes of N. coriiceps were significantly lower in muscle tissue than in liver, kidney and gonad; the livers of males and gonads of females had the highest levels. The differences between the sexes were significant for Zn and Mn in gonads and for Cu in liver. Despite the modest size-range of the specimens, the Cu levels in liver, muscle and kidney were significantly inversely correlated with female mass (P < 0.05) as was gonad Cu level and male body length. Fe mainly presented the highest concentrations and Cu and Mn the lowest ones. Mean muscle metal levels are similar to those of hake, cod, herring and mackerel; however, the level of Fe was slightly higher. Accumulation may occur for Fe, Zn and Mn in liver, kidney and gonad, which have higher levels than those of most prey. Received: 21 October 1997 / Accepted: 1 February 1998     

Inheritance patterns of enzymes and serum proteins of mallard-black duck hybrids

From 1974 to 1976, a breeding program was used to produce hybrids of black ducks and mallards for the evaluation of inheritance patterns of serum proteins and serum, liver and muscle enzymes. In addition to the crosses designed to produce hybrids, a series of matings in 1975 and 1976 were designed to evaluate inheritance patterns of a hybrid with either a black duck or mallard. At the F₁ level, hybrids were easily distinguished using serum proteins. However, once a hybrid was crossed back to either a mallard or black duck, only 12–23% of the progeny were distinguishable from black ducks or mallards using serum proteins and 23–39% using esterases. Muscle, serum and liver enzymes were similar between the two species.     

A Century of Hybridization: Decreasing Genetic Distance Between American Black Ducks and Mallards

American black ducks (Anas rubripes) and mallards (A. platyrhynchos) are morphologically and behaviorally similar species that were primarily allopatric prior to European colonization of North America. Subsequent sympatry has resulted in hybridization, and recent molecular analyses of mallards and black ducks failed to identify two distinct taxa, either due to horizontal gene flow, homoplasy, or shared ancestry. We analyzed microsatellite markers in modern and museum specimens to determine if the inter-relatedness of mallards and black ducks was an ancestral or recent character. Gst, a measure of genetic differentiation, decreased from 0.146 for mallards and black ducks living before 1940, to 0.008 for birds taken in 1998. This is a significant reduction in genetic differentiation, and represents a breakdown in species integrity most likely due to hybridization. Using modern specimens, we observed that despite a lower incidence of sympatry, northern black ducks are now no more distinct from mallards than their southern conspecifics.     

Enzyme activities in plasma, liver and kidney of black ducks and mallards

Activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST), creatine phosphokinase (CPK), alkaline phosphatase (ALP), and lactate dehydrogenase (LDH) were measured in plasma, liver, and kidney, and gamma-glutamyl transferase (GGT) was measured in liver and kidney of black ducks (Anas rubripes). Activities of ALT, AST, GGT, and ornithine carbamyl transferase (OCT) were assayed in plasma, liver, and kidney of game-farm mallards (Anas platyrhynchos). Appreciable OCT and AST activity occurred in both liver and kidney. Activities of ALT, CPK, ALP and GGT were higher in kidney, while LDH was higher in liver, GGT was detected in plasma from one of four mallards.     

Effects of lead shot ingestion on selected cells of the mallard immune system

The immunologic effects of lead were measured in game-farm mallards (Anas platyrhynchos) that ingested lead shot while foraging naturally, mallards intubated with lead shot, and unexposed controls. Circulating white blood cells (WBC) declined significantly in male mallards exposed to lead by either natural ingestion or intubation, but not females. Spleen plaque-forming cell (SPFC) counts were significantly lower in mallards intubated with lead pellets compared to controls. Declines in WBC and SPFC means with increasing tissue lead concentrations provide further evidence that lead exposure reduced immunologic cell numbers. Hormonal activity and diet may have influenced the immunologic effects of lead exposure in this study.