Assessing Potential Environmental Contamination by Baylisascaris procyonis Eggs from Infected Raccoons in Southern Texas.

Baylisascaris procyonis is a large ascarid of raccoons (Procyon lotor) and is a zoonotic threat. We documented the potential rate a raccoon population can contaminate their environment with B. procyonis eggs. We estimated the population size of raccoons using a 9 × 7 trapping grid of Havahart traps, identified locations of raccoon scats through systematic searches, and enumerated the distance B. procyonis eggs passively travel from site of origin upon scat decay. During an 8-week capture period, the raccoon population was estimated to be 19.6 ± 1.3 raccoons within the 63-ha study area (1 raccoon/3.2 ha). There were 781 defecation sites, of which 744 (95.3%) were isolated sites and 37 (4.7%) were latrine sites. Fifty-three (6.8%) defecation sites occurred in areas associated with human structures (commensal zone). Of the noncommensal sites, 9 (1.2%) and 719 (98.8%) sites were identified as latrine sites and isolated scats, respectively. More latrine sites were located within the commensal zone (p < 0.0001, [Formula: see text]) than proportionately available space. Twenty-five raccoon scats containing B. procyonis eggs were allowed to decay on level bare soil by way of simulated rain events, 13 were allowed to desiccate naturally in the environment, and 12 were allowed to desiccate and, subsequently, experience a simulated 1 cm rain event; eggs were found 49 ± 6, 28 ± 8, and 68 ± 8 cm from the initial scat location, respectively. We calculated that a single B. procyonis-infected raccoon could contaminate 0.03 ± 0.01 ha/year with B. procyonis eggs. Our findings indicate that B. procyonis represents a substantial risk to humans in areas where infected raccoons and humans co-occur.

Multiyear Survey of Coccidia, Cryptosporidia, Microsporidia, Histomona, and Hematozoa in Wild Quail in the Rolling Plains Ecoregion of Texas and Oklahoma, USA.

We developed nested PCR protocols and performed a multiyear survey on the prevalence of several protozoan parasites in wild northern bobwhite (Colinus virginianus) and scaled quail (Callipepla squamata) in the Rolling Plains ecoregion of Texas and Oklahoma (i.e. fecal pellets, bird intestines and blood smears collected between 2010 and 2013). Coccidia, cryptosporidia, and microsporidia were detected in 46.2%, 11.7%, and 44.0% of the samples (n = 687), whereas histomona and hematozoa were undetected. Coccidia consisted of one major and two minor Eimeria species. Cryptosporidia were represented by a major unknown Cryptosporidium species and Cryptosporidium baileyi. Detected microsporidia species were highly diverse, in which only 11% were native avian parasites including Encephalitozoon hellem and Encephalitozoon cuniculi, whereas 33% were closely related to species from insects (e.g. Antonospora, Liebermannia, and Sporanauta). This survey suggests that coccidia infections are a significant risk factor in the health of wild quail while cryptosporidia and microsporidia may be much less significant than coccidiosis. In addition, the presence of E. hellem and E. cuniculi (known to cause opportunistic infections in humans) suggests that wild quail could serve as a reservoir for human microsporidian pathogens, and individuals with compromised or weakened immunity should probably take precautions while directly handling wild quail.

Permeability and Viability of Baylisascaris procyonis Eggs in Southern Texas Soils.

Baylisascaris procyonis is a nematode whose definitive host is the raccoon ( Procyon lotor ). Adult parasites are not particularly pathogenic to raccoons; however, larvae in intermediate hosts can cause visceral, ocular, and neural larva migrans. Humans serve as dead-end hosts, and pathological responses are similar to those found in infected intermediate hosts. Infected raccoons expose intermediate hosts through their feces, which can contain millions of B. procyonis eggs. Our objective was to determine how the quantity and viability of B. procyonis eggs in soil changed over time within different soil texture, moisture, and sun exposure. To examine egg survival and movement through soil we placed 100 B. procyonis eggs on 100 squares in 48 boxes representing a full factorial treatment of soil texture, moisture, and sun level. We monitored egg percolation and survival for 2 yr, removing (at 0, 1, 3, 6, 12, 18, and 24 mo) 5 squares from each box and counting the number of viable eggs at each depth in the soil column. Dry soils were mainly impermeable; even after 2 yr, >60% of B. procyonis eggs remained on the surface of all soil textures. Wet soils were more permeable than dry soils, but even in wet sandy soils where the greatest egg movement occurred, it required 1 yr before 60% of eggs transitioned from the soil surface to the next soil depth. For all soil textures, moistures, sun exposures, and depths, >92% of B. procyonis eggs remained viable after 2 yr in the southern Texas environment. Therefore, high exposure risk exists for potential hosts because B. procyonis eggs remain viable on or near the soil surface for at least 2 yr.

Environmental Temperatures in Southern Texas, USA: Implications for Survival of Baylisascaris procyonis Eggs.

Baylisascaris procyonis is an intestinal nematode of raccoons ( Procyon lotor ). Within intermediate hosts, larvae can cause visceral, ocular, and neural migrans. Humans, especially children, have become infected after ingestion of larvated eggs at raccoon latrines. Eggs of B. procyonis have a thermal death point of 62 C. During 2012, we assessed the likelihood of thermal lethality on B. procyonis eggs in southern Texas. We recorded temperature every 30 min with data loggers placed on the ground in full sun and in the shade, buried 5 cm underground and the ground surface exposed to full sun or in shade, in attics with and without exhaust fans, in woodpiles, in sheds, in tree crevices, and in cars parked in the sun. Such locations represented common raccoon latrine sites or locations where B. procyonis eggs could be found. In addition, data loggers placed about 135 cm above the ground in full sun and in shade were used as controls to acquire ambient temperature. The hottest month was August (maximum 44 C), with 15 d that exceeded 38 C. However, only the car reached the lethal temperature limit, and only for 1 h. Southern Texas has one of the warmest climates in the contiguous US; however, it is unlikely that the southern Texas climate is hot enough to kill B. procyonis eggs.

Blood Parasites of Blue-winged Teal ( Anas discors ) from Two Migratory Corridors, in the Southern USA.

We collected 180 Blue-winged Teal ( Anas discors ) in September and October 2002 from Florida, US (n=100, representing the eastern migratory corridor) and the Louisiana-Texas, US, border (n=80, representing the western migratory corridor) and examined for blood parasites using thin heart-blood smears. Leucocytozoon simondi, Haemoproteus nettionis, and microfilariae were found in 16, 23, and 27 birds, respectively. Prevalence of L. simondi and H. nettionis did not vary by migratory corridor, but the prevalence of microfilariae was higher in the western corridor (23%) than the eastern corridor (9%). No differences in prevalence of L. simondi, H. nettionis, and microfilariae were observed by host age or sex. The mean density of L. simondi and H. nettionis averaged 1.5±0.3 and 2.3±0.4 (±SE per 3,000 erythrocytes), respectively. Ranked abundance models for main and interactive effects of corridor, age, and sex were not statistically significant for L. simondi or H. nettionis. Low prevalence and abundance of hematozoa in early autumn migrants reflects the likelihood of low exposure probabilities of Blue-winged Teal on the breeding grounds, compared to their congeners.

HELMINTH INFECTIONS IN NORTHERN BOBWHITES (COLINUS VIRGINIANUS) FROM A LEGACY LANDSCAPE IN TEXAS, USA.

The Northern Bobwhite ( Colinus virginianus ) has declined across its range. The primary cause of this decline is thought to be habitat loss and fragmentation. However, there is speculation that factors such as parasites may play a role. South Texas recently was designated a Legacy Landscape of National Significance for Northern Bobwhite Conservation and is a region with some of the highest bobwhite densities in the US. Limited studies on bobwhite parasites have been conducted in this crucial landscape. We documented helminth parasites infecting bobwhites in South Texas, identified those that are known to be pathogenic to quail, documented pathologic responses to infection, and evaluated infections related to host intrinsic and extrinsic factors. We examined 209 bobwhites and found nine species of helminths including two known to cause tissue damage in bobwhites: Tetrameres pattersoni and Oxyspirura petrowi. The cecal nematode Aulonocephalus pennula was numerically dominant and had the greatest prevalence, intensity, and abundance. Prevalence and abundance of A. pennula were significantly greater in adult than juvenile bobwhites, whereas host sex was not an important factor. Prevalence of A. pennula was significantly greater during the 2012-13 hunting season than the 2013-14 season. The abundance of A. pennula also was significantly greater in bobwhites with greater mass within each age cohort. This research provides insight regarding the factors that influence helminth infections in bobwhites from South Texas and highlights the importance of broad-scale surveys when assessing helminth infections across large regions.

EYEWORMS (OXYSPIRURA PETROWI) IN NORTHERN BOBWHITES (COLINUS VIRGINIANUS) FROM THE ROLLING PLAINS ECOREGION OF TEXAS AND OKLAHOMA, 2011-13.

The Northern Bobwhite ( Colinus virginianus ) has been steadily declining throughout much of its historic range for decades. The Rolling Plains ecoregion of Texas and western Oklahoma, historically rich with wild Northern Bobwhites and one of the last remaining quail strongholds, also has a declining population. During August and October in 2011-13, 348 Northern Bobwhites from the Rolling Plains were examined for eyeworms (Oxyspirura petrowi). Of these 348 Northern Bobwhites, 144 (41.4%) were infected with 1,018 total eyeworms. Eyeworm abundance (mean±SE) was 2.9±0.4 (range 0-64), with an intensity (mean±SE) of 7.1±0.6. Eyeworm prevalence was significantly higher in adult Northern Bobwhites (58.7%) than in juveniles (35.4%). Recent research suggests that eyeworms have the potential to cause cellular tissue damage to the eye, but it is unknown how these worms affect host survivability. This study further expands the regional distribution of O. petrowi in Northern Bobwhites in the Rolling Plains ecoregion and assesses the prevalence and abundance of infection across host age, host sex, and year. Further research is warranted on the life history of O. petrowi and assessing the impacts of eyeworms on their definitive host at individual and population levels.

Shot Ingestion by Wintering Female Northern Pintails (Anas acuta) in the Texas Coastal Plain, 2012-14.

Historically, lead poisoning through lead shot ingestion was one of the largest health issues affecting waterfowl in North America. Lead shot was banned for use in waterfowl hunting in the US in 1991 and was banned in Canada in 1997. However, biologists need to understand how, and if, lead shot remaining in the environment will continue to impact waterfowl. Our goal was to estimate lead and nontoxic shot consumption by female Northern Pintails (Anas acuta) wintering along the Texas coast. We found shot or metal fragments (or both) in the gizzards of 39 (17%) of 227 female Northern Pintails collected along the Texas coast. Of these, lead shot was found in seven gizzards, steel shot was found in 24 gizzards, and other metal and fragments were found in 20 gizzards. Some females consumed multiple shot types. Overall, shot (lead and nontoxic combined) ingestion rates were similar to those found prior to the lead shot ban in Texas (14%) and Louisiana (17%); however, lead shot ingestion rates were considerably lower, suggesting that it is becoming less available over time. All Northern Pintails that had lead shot in their gizzards were collected from coastal habitats. While it seems that lead shot ingestion by Northern Pintails has decreased since the ban was put in place, monitoring lead shot ingestion rates from different regions will provide insight into its availability in different habitats and under various environmental conditions.

Pathological Response of Northern Bobwhites to Oxyspirura petrowi Infections.

The effects of Oxyspirura petrowi infections in northern bobwhites ( Colinus virginianus ) are not well understood. While studies have reported O. petrowi infections, none has histopathologically examined the eye surface and intraorbital glands to assess cellular-level impacts associated with infection. This study is the first to document the histopathology associated with O. petrowi infections. Oxyspirura petrowi occurred on the eye surface as well as in the conjunctiva, lacrimal ducts, lacrimal glands, and Harderian glands. Histopathology showed infections of O. petrowi caused cellular damage to these tissues, scarring and interstitial keratitis of the cornea, and acinar atrophy of the Harderian gland.

Evidence of an Oxyspirura petrowi epizootic in northern bobwhites (Colinus virginianus), Texas, USA.

We captured 36 Northern Bobwhites (Colinus virginianus) in Mitchell County, Texas in June-September 2013, and examined them for the eyeworm Oxyspirura petrowi. We recovered 334 eyeworms from 28 of 29 adult bobwhites (97%); infections ranged from 1-40 worms and mean (± SD) abundance of 11.9 ± 13.0. Three of seven juveniles were infected, and those infected had one eyeworm each. Prevalence of eyeworms was similar among months. However, mean abundance of eyeworms peaked in July and August (3.3 ± 2.1, 13.5 ± 15.0, and 16.9 ± 15.5), and decreased in September (6.3 ± 3.0). We suggest that several previous studies may underreport prevalence and abundance because in those studies only the eye surface and nictitating membrane were examined, and not eye-associated tissue, ducts, glands, or sinuses.

Gene discovery, evolutionary affinity and molecular detection of Oxyspirura petrowi, an eye worm parasite of game birds.

BACKGROUND: Oxyspirura petrowi appears to be emerging as a nematode parasite that could negatively impact Northern Bobwhite quail individuals and populations within Texas and other regions of the United States. Despite this eye worm's potential importance in the conservation of wild quail, little is known about the general biology and genome composition of O. petrowi. To fill the knowledge gap, we performed a small scale random genome sequence survey, sequenced its 18S rRNA and the intergenic region between the 18S and 28S rRNA genes, studied its phylogenetic affinity, and developed a PCR protocol for the detection of this eye worm. RESULTS: We have generated ~240 kb of genome sequence data derived from 348 clones by a random genome survey of an O. petrowi genomic library. The eye worm genome is AT-rich (i.e., 62.2% AT-content), and contains a high number of microsatellite sequences. The discovered genes encode a wide-range of proteins including hypothetical proteins, enzymes, nematode-specific proteins. Phylogenetic analysis based on 18S rRNA sequences indicate that the Spiruroidea is paraphyletic, in which Oxyspirura and its closely related species are sisters to the filarial nematodes. We have also developed a PCR protocol based on the ITS2 sequence that allows sensitive and specific detection of eye worm DNA in feces. Using this newly developed protocol, we have determined that ~28% to 33% of the fecal samples collected from Northern Bobwhites and Scaled Quail in Texas in the spring of 2013 are O. petrowi positive. CONCLUSIONS: The O. petrowi genome is rich in microsatellite sequences that may be used in future genotyping and molecular fingerprinting analysis. This eye worm is evolutionarily close to the filarial nematodes, implying that therapeutic strategies for filariasis such as Loa loa would be referential in developing treatments for the Thelazoidea parasites. Our qPCR-based survey has confirmed that O. petrowi infection is of potential concern to quail managers in Texas.

Acute effects of aflatoxin on northern bobwhites (Colinus virginianus).

Aflatoxin is a widely occurring and harmful mycotoxin produced by strains of Aspergillus spp. growing on vegetable matter. We investigated the concentration of aflatoxin needed to impair normal physiologic responses and induce acute morbidity and mortality in Northern Bobwhites (Colinus virginianus). Ten wild-caught adult bobwhites (five males and five females) from southern Texas were randomly assigned to each treatment group (0, 100, 500, 1,000, and 2,000 parts per billion (ppb) aflatoxin; n=50). We orally administered 100 µL of aflatoxin, derived from Aspergillus flavus, once per week for 4 wk and monitored bird mass, daily feed consumption, liver histology, and blood chemistries. An in vitro white blood cell proliferation test was conducted using spleen tissue to determine the effect of aflatoxin on the immune system. There was no mortality in the control groups, whereas mortalities occurred in all treatment groups except in the 100 ppb aflatoxin treatment. Immunosuppression, reduction in gamma-globulin, glucose, and gamma-glutamyltransferase blood levels, and abnormal liver histology were observed in aflatoxin-exposed quail. Blood chemistry indicated cellular damage to the liver and kidneys. We concluded that short-term, acute doses of aflatoxin as low as 100 ppb can be detrimental to the health of Northern Bobwhites.

Assessing helminth community structure and patterns in gizzard helminths of blue-winged teal (Anas discors).

Helminth community structure (prevalence and abundance) and patterns (associations and distributions) of gizzard worms were examined in blue-winged teal (Anas discors) migrating through south Texas to their wintering grounds (n = 50) and as they migrated back to their breeding grounds (n = 50). Three species of nematodes (Amidostomum acutum, Epomidiostomum uncinatum, and Streptocara crassicauda) and 1 species of cestode (Gastrotaenia cygni) were recovered, representing 1,164 helminth individuals. Amidostomum acutum was the most prevalent (92%) and abundant (817 individuals; 70% of total worms), while G. cygni was the least prevalent (16%) and abundant (23 individuals; 2%). Ninety-eight blue-winged teal were infected with 1 to 4 species. Amidostomum acutum and E. uncinatum were the most frequently (45%) observed association of 2 or more species, followed by A. acutum, E. uncinatum, and S. crassicauda (19%), and A. acutum and S. crassicauda (14%). Significant positive correlations were mainly between A. acutum and E. uncinatum, whereas negative correlations were found between each of these 2 species and S. crassicauda. Prevalence and abundance of A. acutum and S. crassicauda were not influenced by season, host age, or host sex. However, E. uncinatum varied by season and host age. The percent similarity index indicated a relatively consistent component community pattern of high similarity irrespective of season, host age, and sex. The same helminth species were found in both fall and spring, indicating that other helminth species were not being acquired on the wintering grounds.

Helminth community composition, structure, and pattern in six dove species (Columbiformes: Columbidae) of South Texas.

The helminth community composition and structure of 6 species of columbids residing in south Texas are reported and compared herein. Sixty individuals of the following species, rock pigeons (Columba livia [RP]), mourning doves (Zenaida macroura [MD]), Eurasian collared-doves (Streptopelia decaocto [ECD]), white-winged doves (Zenaida asiatica [WWD]), and common ground doves (Columbina passerina [CGD]), and 48 Inca doves (Columbina inca [ID]) were collected during the summer of 2006 and examined for helminths. Twelve helminth species were found (9 nematodes and 3 cestodes), representing 486 individuals. Nematodes numerically dominated the component community in all host species. Overall, helminth prevalence was similar among host sex within all dove species. However, prevalence of Skrjabinia bonini and Hymenolepis sp. in RPs was significantly different among host age groups (P = 0.01, P = 0.0002, respectively). Likewise, prevalence of Killigrewia delafondi was higher (P = 0.0001) in adult WWDs. Based on percent similarity and Jaccard's coefficient of community indices, helminth component communities were dissimilar, and the number of shared helminth species varied among host species. Data from this study suggest that the environment surrounding preferred host habitat and foraging strategies of each host species is the driving force behind helminth component communities. This study emphasizes the importance of examining co-occurring hosts at both local and regional scales to elucidate helminth community structure and patterns.

Hematozoa and a new haemoproteid species from Cathartidae (New World vulture) in South Carolina.

A survey of turkey vultures (Cathartes aura) in South Carolina revealed the presence of a novel haemoproteid. Haemoproteus catharti n. sp. is described from the slides developed from the blood of this vulture. The new species is a thick, halteridial form with complete margins, and is considered distinct from the other recognized haemoproteids from the diurnal raptors in shape, pigment number, parasite outline, and host-family specificity. While reviewing blood films, a series of unusual immature schizonts of Plasmodium sp. were also observed, and these are illustrated, along with another distinct haemoproteid from old slides of poor quality produced from turkey vulture blood.

Ingestion of lead and nontoxic shot by Green-winged Teal (Anas crecca) and Northern Shovelers (Anas clypeata) from the mid-Gulf Coast of Texas, USA.

Ninety-eight Green-winged Teal (Anas crecca) and 84 Northern Shoveler (Anas clypeata) gizzards were examined for ingested shot. One Northern Shoveler had lead and three had steel shot; 24 teal and 17 shovelers had composite nontoxic shot or nonlead metal fragments. Prevalence of ingested lead appears minimal and consistent with other studies conducted after lead-shot bans.

Helminth Community Dynamics in Populations of Blue-Winged Teal (Anas discors) Using Two Distinct Migratory Corridors.

The influence of spatially distinct host subpopulations on helminth community structure and pattern was examined in a migratory avian host species. Forty helminth species represented by 24,082 individuals were collected from 184 blue-winged teal (Anas discors; BWT) from 2 primary migratory corridors in Florida (eastern migratory corridor; EMC) and Louisiana and Texas (western migratory corridor; WMC). Mean species richness was greater in BWT from the WMC (x̅±SE = 10.2 ± 0.3 species) than the EMC (8.6 ± 0.2). The helminth community from the WMC had higher abundances of 6 common/intermediate species. Corridor helminth communities were similar in species composition but less similar when incorporating abundances of those species. Overlapping distributions of phylogenetically related host species that share generalist helminth species across ecologically similar habitats seem to mitigate the isolating mechanisms that are necessary for the distinct coevolutionary pathways to develop between adjacent corridors.

Lead concentrations of white-winged doves, Zenaida asiatica L., collected in the Lower Rio Grande Valley of Texas, USA.

White-winged dove (Zenaida asiatica L.) exposure to environmentally accessible lead is a management concern. Detectable concentrations of lead were found in 142 (66%) of 214 white-winged doves collected in 2002 and 2003 at two wildlife management units in South Texas. Mean +/- SD lead concentration was 0.70 +/- 1.10 microg/g dry weight and ranged from ND to 11.01 microg/g dw. Lead concentrations were significantly lower in the 2002 samples, but similar between management units, age, and gender. White-winged doves in South Texas are being exposed to lead in the environment with some reaching subclinical to toxic levels.

Environmental contaminants in white-winged doves (Zenaida asiatica asiatica) from the Lower Rio Grande Valley of Texas, USA.

Our study determined if white-winged doves (WWDs; Zenaida asiatica asiatica) breeding in locations where environmentally persistent contaminants occur in the Lower Rio Grande Valley of Texas have levels of contaminants that can hinder reproduction. During summer 2003, 70 (32 males, 38 females) adult WWDs were collected from 8 at-risk sites. Liver tissues were analyzed for ten major and trace elements and muscle tissues were analyzed for 20 organochlorine compounds, 21 polychlorinated biphenyl congeners, and toxaphene. Samples were compared to ten (five male, five female) captive-raised control WWDs. Arsenic, chromium, and lead were not detected or were below the detection limit, whereas copper, zinc, selenium, mercury, nickel, cadmium, and silver were detected in 70 (100%), 70 (100%), 52 (74%), 21 (30%), 4 (6%), 1 (1%), and 1 (1%) WWDs, respectively. None of the detected elements had significant sampling site, bird gender, or Site x Gender interactions, and each occurred at background levels. Toxaphene and polychlorinated biphenyls were not detected, whereas DDE, dieldrin, gamma-BHC, and chlordane were detected in 13 (19%), 5 (7%), 2 (3%), and 1 (1%) field-collected WWDs, respectively, but all concentrations occurred at background levels. Concentrations of elements and compounds in field-collected WWDs did not significantly differ from those of the controls. Correlations between leukocyte counts and detectable contaminants generally indicated no correlation between each percentage of leukocyte type present and levels of selenium, copper, zinc, mercury, or DDE detected. One sample that had 0.072 mug/g dieldrin exhibited a low lymphocyte percentage (66%) out of only five samples. Comparisons could not be made with gamma-BHC, chlordane, or the other elements that had a low number of samples with detectable levels. Overall, concentrations of elements and organochlorines found in WWDs were below levels known to impair reproduction and survival in birds.

Blood parasites in wood storks (Mycteria americana) from the southeastern United States.

Wood storks (Mycteria americana) are endangered throughout their breeding range in the United States. Because of this, researchers have had little opportunity to thoroughly examine the continental wood stork population for endoparasites. The blood protozoan Haemoproteus crumenium has been identified from several populations of wood storks in North America. However, there have been no reports of wood storks being infected with species of Plasmodium or Leucocytozoon. During 2003, 42 nestling wood storks were captured at sites in Georgia and 27 free-ranging wood storks were captured at postbreeding dispersal sites in Mississippi and Louisiana. Two thin blood smears were made from each bird, and they were examined for parasites. Haemoproteus crumenium was found in 5 wood storks (7.2%), whereas Leucocytozoon sp., Plasmodium sp., and microfilariae were not observed. Mean intensity of H. crumenium was 4.4 +/- 0.7 (SE) per 2,000 erythrocytes counted. All infected wood storks were subadult or adult and were from postbreeding dispersal sites in Mississippi.