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Scriptaid and 5-aza-2'deoxycytidine enhanced expression of pluripotent genes and in vitro developmental competence in interspecies Black-footed cat cloned embryos

Somatic cell nuclear transfer offers the possibility of preserving endangered species including the black-footed cat, which is threatened with extinction. The effectiveness and efficiency of somatic cell nuclear transfer (SCNT) depends on a variety of factors, but 'inappropriate epigenetic reprogramming of the transplanted nucleus is the primary cause of the developmental failure of cloned embryos. Abnormal epigenetic events such as DNA methylation and histone modifications during SCNT perturb the expression of imprinted and pluripotent-related genes that, consequently, may result in foetal and neonatal abnormalities. We have demonstrated that pregnancies can be established after transfer of black-footed cat cloned embryos into domestic cat recipients, but none of the implanted embryos developed to term and the foetal failure has been associated to aberrant reprogramming in cloned embryos. There is growing evidence that modifying the epigenetic pattern of the chromatin template of both donor cells and reconstructed embryos with a combination of inhibitors of histone deacetylases and DNA methyltransferases results in enhanced gene reactivation and improved in vitro and in vivo developmental competence. Epigenetic modifications of the chromatin template of black-footed cat donor cells and reconstructed embryos with epigenetic-modifying compounds enhanced in vitro development, and regulated the expression of pluripotent genes, but these epigenetic modifications did not improve in vivo developmental competence.

Reproduction in Domestic Animals

Organochlorine pollutants and population status of least terns in South Carolina

Least Tern nesting colonies on the Cape Romain National Wildlife Refuge and surrounding areas in South Carolina were studied from 1971 through 1975. We located 10 colonies including 6 on the Refuge and 4 on nearby coastal islands. The number of nests in each colony ranged from several up to 220. Least Terns began reproductive activity in April, and the egg-laying period ranged from May to July. The earliest hatching record was 6 June. Reproductive success in most colonies seemed poor. Tidal flooding of eggs, predation of eggs and young, and disturbance by domestic animals and man were responsible for most failures. Residues of DDE, PCB's, and other organochlorine pollutants in the eggs were low and posed no identifiable threat to the Least Terns. DDE residues in eggs declined from 0.63 μg/g in 1972 to 0.33 μg/g in 1975. In contrast, PCB residue trends were erratic; mean residue values were 0.40 μg/g in 1972, 1.08 μg/g in 1974, and 0.62 μg/g in 1975. Eggshell thickness means for 1972, 1974, and 1975 were 2 to 7% lower than the pre-1947 mean; but the differences between means were not statistically significant. There is no evidence of a decline in Least Tern populations in South Carolina over the past 30 years such as observed in many other parts of the range of the species. A number of the current nesting islands seem secure from adverse environmental perturbations, although several colonies are on islands that are in danger of extensive development.

South Carolina

Toxicological effects assessment for wildlife in the 21st Century: Review of current methods and recommendations for a path forward

Model species (e.g., granivorous gamebirds, waterfowl, passerines, domesticated rodents) have been used for decades in guideline laboratory tests to generate survival, growth and reproductive data for prospective Ecological Risk Assessments (ERAs) for birds and mammals, while officially adopted risk assessment schemes for amphibians and reptiles do not exist. There are recognized shortcomings of current in vivo methods as well as uncertainty around the extent to which species with different life histories (e.g., terrestrial amphibians, reptiles, bats) than these commonly used models are protected by existing ERA frameworks. Approaches other than validating additional animal models for testing are being developed, but incorporation of such new approach methodologies (NAMs) into risk assessment frameworks will require robust validations against in vivo responses. This takes time, and the ability to extrapolate findings from non-animal studies to organism- and population-level effects in terrestrial wildlife remains weak. Failure to adequately anticipate and predict hazards could have economic and potentially even legal consequences for regulators and product registrants. In order to be able to use fewer animals or replace them altogether in the long-term, vertebrate use and whole organism data will be needed to provide data for NAMs validation in the short term. Therefore, it is worth investing resources for potential updates to existing standard test guidelines used in the laboratory as well as addressing the need for clear guidance on conduct of field studies. Herein we review the potential for improving standard in vivo test methods and for advancing the use of field studies in wildlife risk assessment, as these tools will be needed into the foreseeable future.

Integrated Environmental Assessment and Management

Disease emergence in birds: Challenges for the twenty-first century

The paper by Hartup et al. (2001) on House Finch ( Carpodacus mexicanus ) conjunctivitis is an example of the rapid geographic spread that can result from disease emergence in naïve populations. That event was neither novel nor transient relative to its occurrence or effects. Disease emergence and reemergence are hallmarks of the latter part of the twentieth century (Center for Disease Control 1994, Levins et al. 1994, DaSilva and Laccarino 1999, Gratz 1999). Current examples involving domestic animals include the problems in Europe with bovine spongiform encephalopathy (BSE, or “mad cow disease”) (Brown 2001) and foot-and-mouth disease (FMD) (Kitching 1999). Human health has been affected by diseases caused by an array of viruses (Morse 1993, Nichol et al. 1993, Murphy and Nathanson 1994), bacteria (Dennis 1998, DaSilva and Laccarino 1999), rickettsia (Walker and Dumier 1996, Azad et al. 1997), protozoans (Tuerrant 1997, Saini et al. 2000), and metazoan parasites (Hildreth et al. 1991, Gubler 1998), as well as other causes. Acquired immune deficiency syndrome (AIDS) has received the most notoriety of those diseases (Hahn et al. 2000, Schwartlander et al. 2000). A similar pattern exists on a global scale for free-ranging wildlife populations (Table 1) (Friend 1994, 1995; Epstein et al. 1998, Daszak et al. 2000). However, in comparison to disease emergence affecting humans and domestic animals, response to emerging diseases of wildlife is generally superficial. We present concepts and data to support our contention that failure to adequately address disease emergence in free-ranging wildlife is resulting in a diminished capability to achieve and sustain desired geographic distributions and population abundance for species of wild birds, including some threatened and endangered avifauna. For clarity, we define disease and disease emergence in the context of our use of those terms because they are the focus of our comments. Disease is any departure from health (Guralnik 1982); that is, dysfunction contributing to physiological, physical, reproductive, behavioral, or other impairment that reduces the probability of survival of individuals. If enough individuals are affected, the collective effects can reduce the sustainability of the population. Although disease can result from exposure to a wide variety of physical, chemical, and biological agents and other conditions, we focus this paper on microbes and parasites and to overt mortality caused by them. Thus, disease effects presented only represent the proverbial “tip of the iceberg” relative to the challenges wild avifauna face from disease. Our perspective of disease emergence expands the earlier definitions of emerging diseases by others (Centers for Disease Control and Prevention 1994, Morse 1995) to include all species. Our comments are defined by the context of disease occurrences that have increased within the past three decades, or threaten to increase in the near future relative to populations affected, geographic distribution, or magnitude of effects.

The Auk

Infectious and parasitic diseases of the gray wolf and their potential effects on wolf populations in North America

Numerous infections and parasitic diseases have been reported for the gray wolf, including more than 10 viral, bacterial, and mycotic disease and more than 70 species of helminths and ectoparasites. However, few studies have documented the role of diseases in population dynamics. Disease can affect wolf populations directly by causing mortality or indirectly by affecting physiological and homeostatic processes, thriftiness, reproduction, behavior, or social structure. In addition, wolves are hosts to diseases that can affect prey species, thus affecting wolf populations indirectly by reducing prey abundance or increasing vulnerability to predation. Diseases such as canine distemper and infectious canine hepatitis are enzootic in wolf populations, whereas rabies occurs in wolves primarily as a result of transmission from other species such as artic and red foxes. Contact between wolves and domestic pets and livestock may affect the composition of diseases in wolves and their effects on wolf populations. Dogs were suspected of introducing lice and canine parovirus to several wolf populations. THe potential for disease to affect wolf populations and other wild and domestic animals should be considered in wolf management plans, particularly in plans for reintroduction of wolves to area within their former range.

Conference Paper

An assessment of current wolf Canis lupus domestication hypotheses based on wolf ecology and behaviour

The dog was the first domesticated animal. Its derivation from grey wolves Canis lupus is important to the study of mammalian domestication, and wolf domestication is an active area of investigation. Recent popular books have promoted a hypothesis that wolves domesticated themselves as opposed to the earliest hypothesis that featured pup collection, adoption, and artificial selection. Continuing research has produced a greater understanding of wolf ecology and behaviour, including new insights into the wolf’s interaction with humans. Several characteristics make the wolf conducive to domestication: its sociality, catholic diet, excellent individual and cultural memory, inbreeding tolerance, varied personalities, and adaptable lifestyle. The wolf’s fear of humans is the main impediment and that alone is a factor strongly disfavouring the self-selection hypothesis. However, collecting young pups from dens and raising them would foster their socialising with humans as pack members. Neither hypothesis explains how wolves undergoing domestication were separated reproductively from their wild relatives, an important condition for domestication. We combine information from the literature with information from our own research on wild wolves, archaeology, and canid morphology. We explain how pup collection and deliberate or incidental selection and encouragement to breed with similarly raised wolves could keep incipient dogs separated reproductively from wild relatives. The key is humans regularly feeding the wolves and keeping only those able to live harmoniously with humans. Well-fed, human-dependent wolves would remain near their food supply and in the company of humans, thus increasing their bonds to humans and vice versa . Outbreeding with wild wolves would thus be avoided. Generation after generation of these human-fed, raised, and selected wolves would become increasingly dependent on humans and shaped by them. The pup-adoption hypothesis presented here is more in keeping with basic wolf ecology and behaviour than the self-domestication hypothesis.

Current Biology

Compounds of emerging concern detected in water samples from potable water and wastewater treatment plants and detected in water and bed-sediment samples from sites on the Trinity River, Dallas, Texas, 2009–13

The population in the Dallas-Fort Worth metropolitan area in northern Texas is rapidly growing, resulting in a rapid increase in the demand for potable water and an increase in the discharge of wastewater treatment plant effluent. An assessment of compounds of emerging concern (CECs) in samples collected at potable water and wastewater treatment plants in Dallas and downstream from Dallas in the Trinity River was completed by the U.S. Geological Survey in cooperation with the City of Dallas, Dallas Water Utilities. CECs are synthetic or naturally occurring chemicals that are not commonly monitored in the environment but can enter the environment and cause known or suspected adverse ecological or human health effects. CECs can enter the environment through nonpoint sources (for example, runoff) and point sources (for example, concentrated animal feeding operations and treated-effluent discharge from wastewater treatment plants), which can increase concentrations of CECs especially in highly populated areas. CECs include pharmaceuticals (prescription and nonprescription), steroidal hormones, stanols, sterols, detergents and detergent metabolites (hereinafter referred to as “detergents”), personal-use products, pesticides, polycyclic aromatic hydrocarbons (PAHs), flame retardants, plasticizers, and other organic compounds used in everyday domestic, agricultural, and industrial applications. The release of CECs to the environment went largely unrecognized until relatively recently. Increased loading of certain CECs to the environment, combined with advancements in laboratory analysis methods that resulted in appreciably lower detection levels, brought greater attention to the release of CECs. In addition, synthesis of new chemicals or changes in use and disposal of existing chemicals can create new sources of CECs. Some CECs are endocrine disrupting compounds (EDCs), which can elicit adverse effects on development, behavior, and reproduction of wildlife and can cause dysfunction of human and wildlife endocrine (hormone) systems. Results of studies in the United States and Europe indicate that CECs, their metabolites, and industrial, agricultural, and household wastewater products are present in the aquatic environment, water treatment plants, and septic systems. CECs, especially pharmaceuticals, are of interest because of their persistence, widespread use, and potential to cause adverse effects in humans and nontargeted organisms. There is also concern that some CECs and EDCs resist degradation of water treatment processes at potable water treatment plants (PWTPs) and wastewater treatment plants (WWTPs) and that treated-effluent discharge could contain compounds that negatively affect biota living in receiving waters. Therefore, CECs and EDCs are more likely to be detected in environmental samples collected near areas of high population density where treated effluent from WWTPs can contribute substantially to receiving waters. The U.S. Geological Survey, in cooperation with the City of Dallas, Dallas Water Utilities, evaluated the occurrence and concentrations of selected CECs in samples collected at PWTPs and WWTPs in Dallas and downstream from the Dallas-Fort Worth metropolitan area in the Trinity River, Texas, from August 2009 to December 2013. Water samples were collected at three PWTP sites, two WWTP sites, and five study sites on the Trinity River; all sites where samples were collected were in or downstream from Dallas. These water samples were analyzed for 120 CECs, including human-health pharmaceuticals (prescription and nonprescription), antibiotics, steroidal hormones, stanols, sterols, detergents, personal-use products (flavors and fragrances), pesticides and repellents, industrial wastewater compounds, disinfection compounds, PAHs, flame retardants, and plasticizers. Additionally, bed-sediment samples were collected at each of the five Trinity River sites. The bed-sediment samples were analyzed for 57 CECs. In general, the water treatment processes at PWTPs and WWTPs were effective at reducing detections and concentrations of CECs to undetectable levels or transforming the compounds into degradates that were not analyzed. There were 14 and 73 CECs detected in raw water and in untreated-influent water at PWTPs and WWTPs, respectively. Of these, 11 of the 14 CECs detected in raw-water samples and 44 of the 73 CECs detected in untreated-influent samples were not detected in finished water or in treated-effluent water samples, respectively, indicating that these compounds were removed or degraded to compounds that were not analyzed. Some CECs, however, are resistant to degradation and were detected in untreated and treated water at PWTPs and at WWTPs. The three CECs detected at PWTPs in raw-water and finished-water samples were tris(dichloroisopropyl)phosphate, benzophenone, and methyl salicylate. At WWTPs, 29 CECs were detected, including carbamazepine, sulfamethoxazole, 4-androstene-3,17- dione , 3-beta-coprostanol, acetyl-hexamethyl-tetrahydronaphthalene (AHTN), hexahydro-hexamethyl-cyclopenta-benzopyran (HHCB), 1,4-dichlorobenzene, tribromomethane, benzophenone, and tris(dichloroisopropyl)phosphate, in untreated and treated water, indicating that treatment processes likely did not remove or degrade these compounds. Of the 23 CECs detected in stream-water samples collected at 5 sites on the Trinity River in or near Dallas, 10 CECs (carbamazepine, sulfamethoxazole, caffeine, 3-beta-coprostanol, cholesterol, HHCB, benzophenone, triethyl citrate, tributyl phosphate, and tris(dichloroisopropyl)phosphate) were detected at all 5 sites. The 10 CECs detected in water samples collected at all 5 sites on the Trinity River were also detected in treated-effluent water at WWTPs. Eleven of the 57 targeted CECs were detected in bed-sediment samples collected at study sites on the Trinity River. Of these 11 CECs, only 2 (beta-sitosterol and cholesterol) were detected in bed-sediment samples at all 5 sites on the Trinity River. Nine of these 11 CECs were not detected in any water-column sample, likely because of the strong hydrophobic characteristics of these compounds. Results from water treatment plants indicate that the water treatment process is less effective for removing or degrading compounds that are engineered to be resistant to degradation. These results also indicate the presence of CECs and EDCs at locations upstream from PWTPs in Dallas. Results from Trinity River main-stem sites indicate that some compounds are naturally attenuated during transport, but a few are persistent throughout the study reach. Many CECs and EDCs are hydrophobic and were only detected in bed sediment, indicating multiple pathways through which CECs can persist in the environment. In general, concentrations of CECs in the Dallas-Fort Worth metropolitan area were similar to those found in metropolitan areas nationwide.

Texas

Context-dependent survival, fecundity and predicted population-level consequences of brucellosis in African buffalo

Chronic infections may have negative impacts on wildlife populations, yet their effects are difficult to detect in the absence of long-term population monitoring. Brucella abortus , the bacteria responsible for bovine brucellosis, causes chronic infections and abortions in wild and domestic ungulates, but its impact on population dynamics is not well understood. We report infection patterns and fitness correlates of bovine brucellosis in African buffalo based on (1) 7 years of cross-sectional disease surveys and (2) a 4-year longitudinal study in Kruger National Park (KNP), South Africa. We then used a matrix population model to translate these observed patterns into predicted population-level effects. Annual brucellosis seroprevalence ranged from 8&middot;7% (95% CI = 1&middot;8&ndash;15&middot;6) to 47&middot;6% (95% CI = 35&middot;1&ndash;60&middot;1) increased with age until adulthood (>6) and varied by location within KNP. Animals were on average in worse condition after testing positive for brucellosis ( F = &minus;5&middot;074, P < 0&middot;0001), and infection was associated with a 2&middot;0 (95% CI = 1&middot;1&ndash;3&middot;7) fold increase in mortality (&chi; 2 = 2&middot;039, P = 0&middot;036). Buffalo in low body condition were associated with lower reproductive success ( F = 2&middot;683, P = 0&middot;034), but there was no association between brucellosis and pregnancy or being observed with a calf. For the range of body condition scores observed in the population, the model-predicted growth rate was &lambda; = 1&middot;11 (95% CI = 1&middot;02&ndash;1&middot;21) in herds without brucellosis and &lambda; = 1&middot;00 (95% CI = 0&middot;85&ndash;1&middot;16) when brucellosis seroprevalence was 30%. Our results suggest that brucellosis infection can potentially result in reduced population growth rates, but because these effects varied with demographic and environmental conditions, they may remain unseen without intensive, longitudinal monitoring.

Kruger National Park

Toxicity of Zectran insecticide to several wildlife species

Zectran® (4-dimethylamino-3,5-xylyl methycarbamate), all insecticide proposed for control of spruce budworm, Choristoneura fumiferana (Clemens), was tested against 16 species of pen-reared or captivity-acclimated animals to determine its toxicity and potential hazard to wildlife. The acute oral LD 60 values in mg/kg were: lesser sandhill crane, Grus canadensis canadensis L.., 1.0-4.5; Canada goose, Branta canadensis L.., 2.64; mourning dove, Zenaidura macroura L.., 2.83; mallard duck, Anas platyrhyndura L.., 3.0; coturnix quail, Coturnix coturnix japonica (Teminck & Schlep get), 3.21; mallard duckling 4.2; ring-necked pheasant, Phasianus colchicus L.., 4.5; house finch, Garpodacus mexicanus (Müller), 4.76; chukar partridge, Alectoris graeca (Meisner), 5.24; domestic pigeon, Columba livia (Gmelin), 6.47; sharp-tailed grouse, Pedioecetes phasianellus L.., 10.0; albino Sprague-Dawley rat, Rattus noruegicis (Fischer), 14.1; domestic goat, Capra aegagnzs L., 15.0-30.0; mule deer, Odocoileus hemionus (Rafinesque), 20.0-30.0; house (English) sparrow, Passer domesticus 1.., 50.4; and bullfrog, Rana catesbeiana (Shaw), 283.0-800.0. In general, birds were more susceptible to single oral doses than mammals and reacted more quickly to intoxication and were more quickly affected. In both groups, the first easily recognized symptom of Zectran poisoning was an increase in the respiratory rate. Deaths appeared to occur from respiratory failure following tachypnea and Cheyne-Stokes respiration. Zectran did not show any marked cumulative effect in subacute oral tests with mallards, chukars, and mule deer. The animals were able to tolerate up to about 40% of a single oral LD 50 dose per day for 30 days, and there was some evidence of a low degree of acquired resistance. Zectran applied dermally to albino New Zealand rabbits, Oryctolagus cuniculus (Lilljeborg), at 2000 mg/kg produced no discernible effects. The reproduction of chukars surviving repeated daily doses of Zectran was similar to that of controls. Low application rates, rapid degradation, low dermal toxicity, and the low degree of cumulative toxic action indicate that little hazard to wildlife should result under field conditions, even though Zectran is highly toxic in acute oral tests.

Journal of Economic Entomology

Survival of whirling disease resistant rainbow trout fry in the wild: A comparison of two strains

Introduced pathogens can affect fish populations, and three main factors affect disease occurrence: the environment, host, and pathogen. Manipulating at least one of these factors is necessary for controlling disease. Myxobolus cerebralis , the parasite responsible for salmonid whirling disease, became established in Colorado during the 1990s and caused significant declines in wild Rainbow Trout Oncorhynchus mykiss populations. Attempts to re-establish Rainbow Trout have focused on manipulating salmonid host resistance. A Rainbow Trout strain known as GR × CRR was developed for stocking in Colorado by crossing a whirling-disease-resistant strain known as the German Rainbow Trout (GR) with the Colorado River Rainbow Trout (CRR). The GR × CRR fish exhibit resistance similar to that shown by GR, and survival and reproduction were expected to be similar to those of CRR. One disadvantage of stocking GR × CRR is that outcrossing and backcrossing could decrease resistance, and laboratory studies have indicated that this can occur. A potential disadvantage of stocking pure GR is lower survival due to domestication. To compare fry survival between the strains, a field experiment was conducted in 1.6-km reaches of nine Colorado streams. Each stream was stocked in August 2014 with 5,000 GR × CRR and 5,000 GR individuals. In October 2014, April 2015, and August 2015, apparent survival was assessed. Two laboratory predation experiments were also conducted. The field experiment revealed that short-term apparent survival was influenced by stream, and growth rate was influenced by strain and stream. However, after 12 months, there was no difference in apparent survival or growth rate between the GR and GR × CRR strains. Laboratory experiments showed that survival did not differ between the strains when confronted with Brown Trout Salmo trutta predation. Our results indicate that the GR strain is a viable option for stocking in streams where M. cerebralis is enzootic. Further evaluation is needed to determine whether GR fish will survive to maturity and reproduce.

Journal of Aquatic Animal Health