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USGS2015

Special considerations for specimen collections that may be involved in law enforcement cases

Causes of mortality in wildlife include natural conditions—such as the viral, bacterial, and fungal diseases discussed in other chapters of this manual—and human intervention. Direct human intervention in wildlife deaths may be associated with individual human actions, such as gunshot or poisonings, or with institutions, such as wind farms or mining operations. Mortality that can be directly attributed to humans may be considered a crime based on the animal(s) affected and (or) the agent used. Wildlife laws were established to protect our nation’s wildlife resources and assist in conserving healthy populations of native wildlife. A variety of laws pertain to wildlife, such as the Migratory Bird Treaty Act, which makes it illegal, except as permitted, to capture, kill, or possess a portion or all of any migratory bird, a member of the group that includes the vast majority of wild birds present in the United States. Similar to pathogens that can spread unchecked through a wild population, a person or an institution that illegally kills wildlife may continue this practice indefinitely or until the illegal activities are detected and addressed. Convicting the perpetrator of a crime, however, requires that certain practices and procedures be followed so that a solid legal case can be constructed. As with any crime, investigation of the event is led by law enforcement personnel who are well-versed in the applicable laws, oversee the collection of evidence, and pursue persons or entities of interest. These individuals are uniquely qualified and trained to maintain the integrity of evidence, evaluate suspects to effectively prosecute the case, and effect the penalties of the law. The investigation of direct crimes against humans may involve teams of 5–10 law enforcement agents and specialists—each with a certain focus such as blood spatter, fingerprints, or ballistics—in processing a crime scene. In contrast, investigations of crimes against wildlife generally involve markedly fewer wildlife law enforcement agents, and in many instances of unlawful wildlife death, only one law enforcement agent is available to process an entire crime scene and work on a case. Thus, the assistance of individuals familiar with the geographical area, the endemic animal populations, and the local ecology can be very important for building an effective case. The aims of this chapter are to 1) help biologists or field personnel recognize the signs that indicate a crime may have been committed, and 2) to give readers a basic understanding of how a wildlife crime scene is processed. This chapter is not intended to give readers the tools to investigate a crime scene alone, but will hopefully make them a valuable asset to law enforcement agents who respond to the scene. An effectively processed case, resulting in prosecution of a suspect, supports the law as a deterrent to future acts that could endanger the conservation of wildlife.

Techniques and Methods↗

USGS2019

Track tube construction and field protocol for small mammal surveys with emphasis on the endangered Pacific pocket mouse (Perognathus longimembris pacificus)

Track tubes are used to identify small animals by their tracks. Animals that are small enough to fit into the tubes walk over ink pads and onto cardstock paper to obtain bait within the tube, leaving their footprints. The tracking tubes described in this document are designed to be set on the ground with free access and exit at either end with additional design components for stability, durability, and efficiency. They are also designed to prevent dirt from getting onto the ink pads and to decrease the ability of birds and other mammals to pull out track cards or bait. We describe detailed methods for constructing, setting and checking track tubes, as well as measuring and identifying small mammal prints for a small mammal study. The protocols described are for monitoring the Pacific pocket mouse (PPM); however, this method can be applied to many small mammal species that have uniquely identifiable tracks in relation to co-occurring species. We have deployed track tubes for over 5 years on Marine Corps Base Camp Pendleton for PPM discovery efforts and to monitor the three extant PPM populations on Base. We have shown that nightly detection probability is similar to that of live-trapping, but the track tubes can be checked weekly or bi-monthly. We use this passive and economical method to assess timing of annual emergence and torpor, seasonal activity, and localized colonization and extinction events. Using this method, we can model occupancy dynamics in relation to habitat and disturbance covariates that directly inform management and support a monitoring and management feedback loop for this species.

California↗

USGS2000

Monthly variability and possible sources of nitrate in ground water beneath mixed agricultural land use, Suwannee and Lafayette Counties, Florida

In an area of mixed agricultural land use in Suwannee and Lafayette Counties of northern Florida, water samples were collected monthly from 14 wells tapping the Upper Floridan aquifer during July 1998 through June 1999 to assess hydrologic and land-use factors affecting the variability in nitrate concentrations in ground water. Unusually high amounts of rainfall in September and October 1998 (43.5 centimeters total for both months) resulted in an increase in water levels in all wells in October 1998. This was followed by unusually low amounts of rainfall during November 1998 through May 1999, when rainfall was 40.7 centimeters below 30-year mean monthly values. The presence of karst features (sinkholes, springs, solution conduits) and the highly permeable sands that overlie the Upper Floridan aquifer provide for rapid movement of water containing elevated nitrate concentrations to the aquifer. Nitrate was the dominant form of nitrogen in ground water collected at all sites and nitrate concentrations ranged from less than 0.02 to 22 milligrams per liter (mg/L), as nitrogen. Water samples from most wells showed substantial monthly or seasonal fluctuations in nitrate concentrations. Generally, water samples from wells with nitrate concentrations higher than 10 mg/L showed the greatest amount of monthly fluctuation. For example, water samples from six of eight wells had monthly nitrate concentrations that varied by at least 5 mg/L during the study period. Water from most wells with lower nitrate concentrations (less than 6 mg/L) also showed large monthly fluctuations. For instance, nitrate concentrations in water from four sites showed monthly variations of more than 50 percent. Large fluctuations in nitrate concentrations likely result from seasonal agricultural practices (fertilizer application and animal waste spreading) at a particular site. For example, an increase in nitrate concentrations observed in water samples from seven sites in February or March 1999 most likely results from application of synthetic fertilizers during the late winter months. Lower nitrate concentrations were detected in water samples from five of eight wells sampled during high-flow conditions for the Suwannee River in March 1998 compared to low-flow conditions in November 1998. Evidence for reduction of nitrate due to denitrification reactions was observed at one site (AC-1), as indicated by elevated concentrations of nitrogen gas and a corresponding increase in nitrogen isotope (d15N-NO3) values with a decrease in nitrate concentrations. Denitrification is unlikely at other sites based on the presence of dissolved oxygen concentrations greater than 2 mg/L in ground water and no observed trend between nitrate concentrations and values d15N-NO3 values. Nitrate was the dominant nitrogen species in most monthly rainfall samples; however, ammonium concentrations were similar or greater than nitrate during November and December 1998. During February through May 1999, both nitrate and ammonium concentrations were substantially higher in monthly rainfall samples collected at the study area compared to mean monthly concentrations at the Bradford Forest site located east of the study area, which is part of the National Atmospheric Deposition Program/National Trends Network. Also, higher nitrogen deposition rates in the study area compared to those at Bradford Forest could indicate that substantial amounts of ammonia are volatilized from fertilizers and animal wastes, released to the atmosphere, and incorporated as nitrate and ammonium in rainfall deposited in the middle Suwannee River Basin. Ground-water samples from most sites had d15N-NO3 values that indicated a mixture of inorganic and organic sources of nitrogen, which corresponded to multiple land uses where both synthetic fertilizers and manure are used on fields near these sites. Distinct d15N-NO3 signatures, however, were observed at some sites. For example, water samples from areas of row-crop farming as the dominant land use had d15N-NO3 values less than 4 per mil, indicating an inorganic nitrogen source such as synthetic fertilizer. In contrast, d15N-NO3 values greater than 9 per mil were found in water samples from three sites where manure from dairy and or poultry operations is spread on fields throughout the year.

Florida↗

USGS1990

Hydrology and land use in Grand Traverse County, Michigan

Glacial deposits are the sole source of ground-water supplies in Grand Traverse County. These deposits range in thickness from 100 to 900 feet and consist of till, outwash, and materials of lacustrine and eolian origin. In some areas, the deposits fill buried valleys that are 500 feet deep. Sedimentary rocks of Paleozoic age, which underlie the glacial deposits, are mostly shale and are not used for water supply. Of the glacial deposits, outwash and lacustrine sand are the most productive aquifers. Most domestic wells obtain water from sand and gravel at depths ranging from 50 to 150 feet and yield at least 20 gallons per minute. Irrigation, municipal, and industrial wells capable of yielding 250 gallons per minute or more are generally greater than 150 feet deep. At places in the county where moranial deposits contain large amounts of interbedded silt and clay, wells are generally deeper and yields are much lower. Areal variations in the chemical and physical characteristics of ground and surface water are related to land use and chemical inputs to the hydrologic system. Information on fertilizer application, septic-tank discharges, animal wastes, and precipitation indicate that 40 percent of nitrogen input is from precipitation, 6 percent from septic tanks, 14 percent from animal wastes, and 40 percent from fertilizers. Streams and lakes generally have a calcium bicarbonate-type water. The dissolved-solids concentration of streams ranged from 116 to 380 milligrams per liter, and that of lakes, from 47 to 170 milligrams per liter. Water of streams is hard to very hard; water of lakes ranges from soft to hard. The maximum total nitrogen concentration found in streams was 4.4 milligrams per liter. Water of lakes have low nitrogen concentrations; the median nitrate concentration is less than 0.01 milligrams per liter. Pesticides (Parathion and Simazine) were detected in low concentrations at six stream sites; 2,4-D was detected in low concentrations in water of two lakes. Relationships between land use and the yield of dissolved and suspended substances could not be established for most stream basins. Calcium and bicarbonate are the principal dissolved substances in ground water. Dissolved-solids concentrations ranged from 70 to 700 milligrams per liter; the countywide mean concentration is 230 milligrams per liter. The mean nitrate concentration is 1.3 milligrams per liter; about 1.6 percent of the county's ground water has nitrate concentrations that exceed the U.S. Environmental Protection Agency's maximum drinking water level of 10 milligrams per liter. An effect of fertilizer applications on ground-water quality is evident in some parts of the county.

Michigan↗

USGS1995

Hydrology and the effects of selected agricultural best-management practices in the Bald Eagle Creek watershed, York County, Pennsylvania, prior to and during nutrient management: Water-quality study for the Chesapeake Bay Program

The U.S. Geological Survey, in cooperation with the Susquehanna River Basin Commission and the Pennsylvania Department of Environmental Resources, conducted a study as part of the U.S. Environmental Protection Agency's Chesapeake Bay Program to determine the effects of nutrient management of surface-water quality by reducing animal units in a 0.43-square-mile agricultural watershed in York County. The study was conducted primarily from October 1985 through September 1990 prior to and during the implementation of nutrient-management practices designed to reduce nutrient and sediment discharges. Intermittent sampling continued until August 1991. The Bald Eagle Creek Basin is underlain by schist and quartzite. About 87 percent of the watershed is cropland and pasture. Nearly 33 percent of the cropland was planted in corn prior to nutrient management, whereas 22 percent of the cropland was planted in corn during the nutrient-management phase. The animal population was reduced by 49 percent during nutrient management. Average annual applications of nitrogen and phosphorus from manure to cropland were reduced by 3,940 pounds (39 percent) and 910 pounds (46 percent), respectively, during nutrient management. A total of 94,560 pounds of nitrogen (538 pounds per acre) and 26,400 pounds of phosphorus (150 pounds per acre) were applied to the cropland as commercial fertilizer and manure during the 5-year study. Core samples from the top 4 feet of soil were collected prior to and during nutrient management and analyzed from concentrations of nitrogen and phosphorus. The average amount of nitrate nitrogen in the soil ranged from 36 to 135 pounds per acre, and soluble phosphorus ranged from 0.39 to 2.5 pounds per acre, prior to nutrient management. During nutrient management, nitrate nitrogen in the soil ranged from 21 to 291 pounds per acre and soluble phosphorus ranged from 0.73 to 1.7 pounds per acre. Precipitation was about 18 percent below normal and streamflow was about 35 percent below normal prior to nutrient management, whereas precipitation was 4 percent above normal and streamflow was 3 percent below normal during the first 2 years of nutrient management. Eighty-four percent of the 20.44 inches of streamflow was base flow prior to nutrient management and 54 percent of the 31.14 inches of streamflow was base flow during the first 2 years of the nutrient-management phase. About 31 percent of the measured precipitation during the first 4 years of the study was discharged as surface water; the remaining 69 percent was removed as evapotranspiration or remained in ground-water storage. Median concentrations of total nitrogen and dissolved nitrate plus nitrite in base flow increased from 4.9 and 4.1 milligrams per liter as nitrogen, respectively, prior to nutrient management to 5.8 and 5.0 milligrams per liter, respectively, during nutrient management. Median concentrations of ammonia nitrogen and organic nitrogen did not change significantly in base flow. Median concentrations of total and dissolved phosphorus in base flow did not change significantly and were 0.05 and 0.03 milligrams per liter as phosphorus, respectively, prior to the management phase, and 0.05 and 0.04 milligrams per liter, respectively, during the management phase.

Pennsylvania↗

USGS1994

Nitrate in ground water and spring water near four dairy farms in North Florida, 1990-93

Concentrations of nitrate and other selected water- quality characteristics were analyzed periodically for two years in water from 51 monitoring wells installed at four farms and in water discharging from three nearby springs along the Suwannee River in Lafayette and Suwannee Counties to examine the quality of ground water at these farms and the transport of nutrients in ground water to the nearby spring-fed Suwannee River: Ground water from shallow wells, which were completed in the top ten feet of the saturated zone in a surficial sandy aquifer and in the karstic Upper Floridan aquifer generally had the highest concentrations of nitrate, ranging from <.02 to 130 mg/L as nitrogen. Nitrate concentrations commonly exceeded the primary drinking water standard of 10 mg/L for nitrate as nitrogen in water from shallow wells, which tapped the top ten feet of the uppermost aquifers near waste-disposal areas such as wastewater lagoons and defoliated, intensive-use areas near milking barns. Upgradient from waste-disposal areas, concentrations of nitrate in ground water were commonly less than 1 mg/L as nitrogen. Water samples from deep wells (screened 20 feet deeper than shallow wells in these aquifers) generally had lower concentrations of nitrate (ranging from <0.02 to 84 mg/L) than water from shallow wells. Water samples from the three monitored springs (Blue, Telford, and Convict Springs) had nitrate concentrations ranging from 1.5 to 6.5 mg/L as nitrogen, which were higher than those typically occurring in water from upgradient wells at the monitored dairy farms or from back- ground wells sampled in the region. Analyses of nitrogen isotope ratios in nitrate indicated that leachate from animal wastes was the principal source of nitrate in ground water adjacent to waste-disposal areas at the monitored and unmonitored dairy farms. Leachate from a combi- nation of fertilizers, soils, and animal wastes appeared to be the source of nitrate in ground- water downgradient from pastures and wastewater spray fields at dairy farms and in water discharging from three nearby springs. Although denitrifying bacteria were present in counts sometimes exceeding 240,000 colonies/100mL in water from dairy-farm monitoring wells, ground water in the uppermost aquifers in Lafayette and Suwannee Counties generally contained too much oxygen for denitrification to remove nitrate from shallow ground water. Denitrification was more likely to occur in deeper ground water, which typically has lower dissolved oxygen concentrations.

Florida↗

USGS1995

Presence, distribution, and potential sources of nitrate and selected pesticides in the surficial aquifer along the Straight River in north-central Minnesota, 1992-93

The presence and distribution of nitrate and selected pesticides in ground water in the surficial aquifer along the Straight River in north-central Minnesota were studied. Local residents and public officials are concerned that these substances may pose a health hazard to humans and livestock. Nitrate and pesticides may move downward from cultivated croplands, livestock feedlots and manured fields, waste-water lagoons, and residential development to the ground water. Ground water near the water table ranged in nitrate-nitrogen concentration (based on median values determined for sampled monitoring wells) from less than 5 to a little greater than 20 mg/L (milligrams per liter) except for one concentration of 50 mg/L downgradient from a feedlot and manured field. Increased nitrate-nitrogen concentrations generally were coincident with cultivated croplands. Decreased nitrate-nitrogen concentrations generally were coincident with forests. Trace amounts of atrazine were detected in 4 of 8 ground-water samples collected from 8 monitoring wells screened near the water table. Detections were more frequent in cultivated croplands (detections at 3 of 5 monitoring wells) than in forests (detections at 1 of 3 monitoring wells). Atrazine concentrations ranged from 0.01 to 0.11 ( μ g/L (micrograms per liter), which are well below the MCL (Maximum Contaminant Level) of 3 μ g/L established by the USEPA (U.S. Environmental Protection Agency). Trace amounts of metolachlor and alachlor were detected in one of the samples in which atrazine was detected. MCLs for metolachlor and alachlor have not been established by the USEPA. The median nitrate-nitrogen concentration determined from sampled monitoring wells along the direction of ground-water flow through five land-use settings: (1) increased from slightly greater than 0 to 50 mg/L near the water table at a feedlot and an adjacent manured field; animal waste was a nitrate source; (2) increased from 8.0 to 16 mg/L near the water table at cultivated croplands irrigated with municipal treated waste water; fertilizer probably was a nitrate source; (3) increased from slightly greater than 0 to 14 mg/L near the bottom of the aquifer at cultivated croplands irrigated with ground water; fertilizer probably was a nitrate source; (4) increased from 1.6 to 3.7 mg/L near the water table and from slightly greater than 0 to 2.8 mg/L near the bottom of the aquifer at a residential development; septic-system leachates, and possibly lawn fertilizer, may have been nitrate sources; and (5) decreased from 14 to 2.6 mg/L near the water table at three waste-water treatment lagoons; waste water was not a nitrate source. Upgradient to downgradient mean or individual nitrogen isotope δ 15 N values in % o (delta units in parts per thousand) determined for sampled monitoring wells along the direction of ground-water flow through the five land-use settings were: (1) 5.1 % o and 4.0 % o for the feedlot and adjacent manured field; (2) 1.1 % o and 0.9 % o for the cultivated croplands irrigated with waste water; (3) 3.8 % o and 2.7 % o for the cultivated croplands irrigated with ground water; (4) 3.4 % o and 4.9 % o for the residential development; and (5) 1.7 % o and 3.0 % o for the three waste-water lagoons. Nitrate from fertilizer appeared to have been present in ground water at the waste-water lagoons, cultivated croplands irrigated with waste water, and cultivated croplands irrigated with ground water. Nitrate from soil organic matter rather than from animal waste appeared to have been present in ground water at the feedlot and adjacent manured field.

Minnesota↗

USGS1997

A comparison of drainage basin nutrient inputs with instream nutrient loads for seven rivers in Georgia and Florida, 1986-90

Instream nutrient loads of the Altamaha, Suwannee, St. Johns, Satilla, Ogeechee, Withlacoochee, and Ochlockonee River Basins were computed and compared with nutrient inputs for each basin for the period 1986-90. Nutrient constituents that were considered included nitrate, ammonia, organic nitrogen, and total phosphorus. Sources of nutrients considered for this analysis included atmospheric deposition, fertilizer, animal waste, wastewater-treatment plant discharge, and septic discharge. The mean nitrogen input ranged from 2,400 kilograms per year per square kilometer (kg/yr)km2 in the Withlacoochee River Basin to 5,470 (kg/yr)km2 in the Altamaha River Basin. The Satilla and Ochlockonee River Basins also had large amounts of nitrogen input per unit area, totaling 5,430 and 4,920 (kg/yr)km2, respectively. Fertilizer or animal waste, as sources of nitrogen, predominated in all basins. Atmospheric deposition contributed less than one-fourth of the mean total nitrogen input to all basins and was consistently the third largest input in all but the Ogeechee River Basin, where it was the second largest. The mean total phosphorus input ranged from 331 (kg/yr)km2 in the Withlacoochee River Basin to 1,380 (kg/yr)km2 in both the Altamaha and Satilla River Basins. The Ochlockonee River Basin had a phosphorus input of 1,140 (kg/yr)km2. Per unit area, the Suwannee River discharged the highest instream mean total nitrogen and phosphorus loads and also discharged higher instream nitrate loads per unit area than the other six rivers. Phosphorus loads in stream discharge were highest in the Suwannee and Ochlockonee Rivers. The ratio of nutrient outputs to inputs for the seven studied rivers ranged from 4.2 to 14.9 percent, with the St. Johns (14.9 percent) and Suwannee (12.1 percent) Rivers having significantly higher percentages than those from the other basins. The output/input percentages for mean total phosphorus ranged from 1.0 to 7.0 percent, with the St. Johns (6.2 percent) and Suwannee (7.0 percent) Rivers exporting the highest percentage of phosphorus. Although instream nutrient loads constitute only one of the various pathways nutrients may take in leaving a river basin, only a relatively small part of nutrient input to the basin leaves the basin in stream discharge for the major coastal rivers examined in this study. The actual amount of nutrient transported in a river basin depends on the ways in which nutrients are physically handled, geographically distributed, and chemically assimilated within a river basin.

Florida, Georgia↗

USGS1994

Membrane-micelle model for humus in soils and sediments and its relation to humification

Humification, the process whereby biomass consisting of dead plant and animal remains is converted into soil organic matter (humus), is one of the basic processes of the carbon cycle. The organic compounds that make up plant and animal tissue are thermodynamically unstable in the oxidizing atmosphere at the surface of the Earth. After the organisms in which they are incorporated die, the compounds are converted back to carbon dioxide and water by degradation reactions catalyzed by enzymes secreted by micro-organisms. However, not all the organic compounds in the dead biomass are immediately converted; some of the material is only partially oxidized. The residue left after partial oxidative degradation of the dead biomass is the source of the organic compounds that accumulate in soils and sediments as humus. Previously, humification was thought to involve a conversion of degradation products by a series of polymerization reactions into new types of polymeric species that are different from the precursor molecular species in the original biomass. However, it is proposed here that the depolymerization and oxidation reactions that take place during the enzymatic degradation of biopolymers produce amphiphiles--molecules that have a polar (hydrophilic) part and a nonpolar (hydrophobic) part. These amphiphiles that result from the partial oxidative degradation of dead biomass assemble spontaneously into ordered aggregates in which the hydrophobic parts of the molecules form the interiors and the hydrophilic parts of the molecules make up the exterior surfaces of the aggregates. These ordered aggregates constitute the humus in soils and sediments. Humus ordered aggregates most likely exist as bilayer membranes coating mineral grains and as micelles in solution.

Water Supply Paper↗

USGS2009

Attempts to identify Clostridium botulinum toxin in milk from three experimentally intoxicated Holstein cows

Three adult lactating Holstein cows were injected in the subcutaneous abdominal vein with 175 ng/kg of body weight of Clostridium botulinum type C toxin (451 cow median toxic doses) to determine if this botulinum toxin crosses the blood–milk barrier. Whole blood (in sodium heparin) and clotted blood serum samples were taken at 0 min, 10 min, and 3, 6, 9, and 12 h postinoculation. Milk samples were taken at 0 min and at 3, 6, 9 and 12 h postinoculation. All samples were tested for the presence of the toxin using the mouse bioassay and immunostick ELISA test. The immunostick ELISA identified the toxin in whole blood and the mouse bioassay identified the toxin in serum at all times examined in all 3 animals. Toxin was not identified by either detection method in milk samples collected from the 3 animals. From these results, it appears that Clostridium botulinum type C toxin does not cross from the blood to the milk in detectable concentrations.

Journal of Dairy Science↗

USGS2025

Pit tag application in native freshwater mussels: Case studies across small, medium, and large rivers

Since their first use in the mid-1980s, external passive integrated transponder (PIT) tags have facilitated innovative investigations into multiple biological traits of animals. For native freshwater mussels, PIT tags are frequently used in capture-mark-recapture applications because they allow repeated, noninvasive sampling, are easy to apply, have high retention rates, and have negligible short-term effects on growth and survival. Because of these traits, resource managers and scientists are using PIT-tagged animals to estimate survival and movement of mussels associated with restoration efforts. However, consistency is limited in how PIT tags are affixed, monitored, and reported. Thus, our objectives were to (1) share our collective experiences in PIT tagging mussels across three case studies in small, medium, and large rivers and (2) propose guidelines for tagging and reporting data from PIT tag studies with native freshwater mussels to facilitate comparisons across future studies. The number of studies that have marked mussels with PIT tags has increased over the past 10 years. The ability to detect mussels using PIT tags has substantially advanced research in three areas of mussel ecology: (1) estimating vital rates (e.g., growth and survival), (2) tracking movements and behaviors of captively propagated, wild, and translocated individuals, and (3) improving our understanding of life history traits, such as reproductive timing. Each case study offers insights on tagging methods, tag loss, tag retention, and monitoring frequency across multiple species that range in conservation status from common to rare. We conclude with best-practice guidelines for placing PIT tags on freshwater mussels and a list of variables that could be reported in future studies to facilitate cross-system comparisons.

Freshwater Mollusk Biology and Conservation↗

USGS2020

Cytology reveals diverse cell morphotypes and cellin-cell interactions in normal collector sea urchins Tripneustes gratilla

Echinoderms such as sea urchins are important in marine ecosystems, particularly as grazers, and unhealthy sea urchins can have important ecological implications. For instance, unexplained mortalities of Diadema antillarum in the Caribbean were followed by algal overgrowth and subsequent collapse of coral reef ecosystems. Unfortunately, few tools exist to evaluate echinoderm health, making management of mortalities or other health issues problematic. Hematology is often used to assess health in many animal groups, including invertebrates, but is seldom applied to echinoderms. We used a standard gravitometric technique to concentrate fixed coelomocytes from the collector sea urchin Tripneustes gratilla onto microscope slides, permitting staining and enumeration. Using Romanowsky stain and electron microscopy to visualize cell details, we found that urchin cells could be partitioned into different morphotypes. Specifically, we enumerated phagocytes, phagocytes with perinuclear cytoplasmic dots, vibratile cells, colorless spherule cells, red spherule cells, and red spherule cells with pink granules. We also saw cell-in-cell interactions characterized by phagocytes apparently phagocytizing mainly the motile cells including red spherule cells, colorless spherule cells, and vibratile cells disproportionate to underlying populations of circulating cells. Cell-in-cell interactions were seen in 71% of sea urchins, but comprised <1% of circulating cells. Finally, about 40% of sea urchins had circulating phagocytes that were apparently phagocytizing spicules. The coelomic fluid collection and slide preparation methods described here are simple, field portable, and might be a useful complementary tool for assessing health of other marine invertebrates, revealing heretofore unknown physiological phenomena in this animal group.

Hawaii↗

USGS2023

Mass mortality of collector urchins Tripneustes gratilla in Hawai`i

As grazers, sea urchins are keystone species in tropical marine ecosystems, and their loss can have important ecological ramifications. Die-offs of urchins are frequently described, but their causes are often unclear, in part because systematic examinations of animal tissues at gross and microscopic level are not done. In some areas, urchins are being employed to control invasive marine algae. Here, we describe the pathology of a mortality event in Tripneustes gratilla in Hawai`i where urchins were translocated to control invasive algae. Although we did not determine the cause of the mortality event, our investigation indicates that animals died from inflammation of the test and epidermal ulceration, followed by inability to maintain coelomic fluid volume, colonization of coelomic fluid by opportunists (diatom, algae), and inappetence. Parasites, bacteria, fungi, and viruses were not evident as a primary cause of death. Pathology was suggestive of a toxin or other environmental cause such as lack of food, possibilities that could be pursued in future investigations. These findings highlight the need for caution and additional tools to better assess health when translocating marine invertebrates to ensure maximal biosecurity.

Hawaii↗

USGS2003

Bacteremia in free-ranging Hawaiian green turtles, Chelonia mydas, with fibropapillomatosis

Past studies of free-ranging green turtles Chelonia mydas with fibropapillomatosis (FP) in Hawaii have shown that animals become immunosuppressed with increasing severity of this disease. Additionally, preliminary clinical examination of moribund turtles with FP revealed that some animals were also bacteraemic. We tested the hypothesis that bacteraemia in sea turtles is associated with the severity of FP. We captured free-ranging green turtles from areas in Hawaii where FP is absent, and areas where FP has been endemic since the late 1950s. Each turtle was given an FP severity score ranging from 0 (no tumours) to 3 (severely affected). A fifth category included turtles that were stranded ashore and moribund with FP. We found that the percentage of turtles with bacteraemia increased with the severity of FP, and that the majority of bacteria cultured were Vibrio spp. Turtles with severe FP were more susceptible to bactaeremia, probably in part due to immunosuppression. The pattern of bacteraemia in relation to severity of disease strengthens the hypothesis that immunosuppression is a sequel to FP.

Hawai'i↗

USGS2015

Stable carbon and nitrogen isotope trophic enrichment factors for Steller sea lion vibrissae relative to milk and fish/invertebrate diets

Nutritional constraints have been proposed as a contributor to population declines in the endangered Steller sea lion Eumetopias jubatus in some regions of the North Pacific. Isotopic analysis of vibrissae (whiskers) is a potentially useful approach to resolving the nutritional ecology of this species because long-term (up to 8 yr) dietary information is sequentially recorded and metabolically inert once formed. Additionally, vibrissae are grown in utero , potentially offering indirect inference on maternal diet. However, diet reconstruction using isotopic techniques requires a priori knowledge of trophic enrichment factors (TEFs), which can vary relative to diet quality and among animal species. In this study, we provide new TEF estimates for (1) maternal relative to pup vibrissae during both gestation and nursing and (2) adult vibrissae relative to a complex diet. Further, we refine vibrissa-milk TEFs based on an additional 76 animals with an age distribution ranging from 1 to 20 mo. Mother-pup vibrissae TEF values during gestation and nursing were near zero for δ 13 C and averaged 0.8 and 1.6&permil;, respectively, for δ 15 N. In contrast, vibrissa-fish/invertebrate TEFs averaged 3.3 (± 0.3 SD) and 3.7&permil; (±0.3) for lipid-free δ 13 C and δ 15 N, respectively. Average lipid-free δ 13 C and δ 15 N vibrissa-milk TEFs were 2.5 (±0.9) and 1.8&permil; (±0.8), respectively, and did not differ among metapopulations. Empirically determined TEFs are critical for accurate retrospective diet modeling, particularly for evaluating the hypothesis of nutritional deficiency contributing to the lack of Steller sea lion population recovery in some regions of Alaska.

Marine Ecology Progress Series↗

USGS2016

Use of noninvasive genetics to assess nest and space use by white-tailed eagles

Movement and space use are important components of animal interactions with the environment. However, for hard-to-monitor raptor species, there are substantial gaps in our understanding of these key determinants. We used noninvasive genetic tools to evaluate the details of space use over a 3-yr period by White-tailed Eagles ( Haliaeetus albicilla ) at the Naurzum Zapovednik in northern Kazakhstan. We genotyped, at 10 microsatellite markers and one mitochondrial marker, 859 eagle feathers and assigned naturally shed feathers to individuals. We identified 124 White-tailed Eagles, including both members of 5–10 pairs per year, and were able to monitor birds across years. Distances between eagle nests and hunting perches were always greater than nearest neighbor distances, eagles never used the closest available hunting perch, and hunting perches were always shared with other eagles. When eagles switched nests between years, the nests they chose were almost always well outside the space that theory predicted they defended the prior year. Our data are inconsistent with classical territorial and colonial models of resource use; they more closely resemble semi-colonial behavior. It is unlikely that standard methods of animal tracking (e.g., marking and telemetry), would have provided a similarly cost-effective mechanism to gain these insights into spatial and temporal aspects of eagle behavior. When combined with existing information on space use of other local species, these data suggest that partitioning of spatial resources among White-tailed Eagles and other eagles at the Zapovednik may be facilitated by the alternative strategies of space use they employ.

Naurzum Zapovednik↗

USGS2025

Northward migrations of nonbreeding Bald Eagles from Arizona, USA.

Knowledge of the spatiotemporal patterns of migratory and nonbreeding-season movements by animals is critical for conservation, but can be difficult to obtain if animals move far from known breeding territories and across administrative and country borders. To understand the migratory movements of Bald Eagles ( Haliaeetus leucocephalus ) originating from a demographically closed population in Arizona, USA, we deployed GPS transmitters on 24 juveniles and 2 nonbreeding adult eagles between 2017 and 2023. We identified common migration routes, stopover locations, and migration phenology. Eagles moved north from Arizona during the population's post-breeding period in the boreal spring and summer, and returned south in autumn in advance of the breeding season. Eagles migrated along two primary routes: (1) along the Wasatch Mountains in Utah and extending into the Rocky Mountains of southern Idaho, and (2) through eastern Nevada and along the Rocky Mountains in western and northern Idaho. Stopover locations were frequently near lakes and rivers. Recent fledglings began migration later than older individuals and 15 of 16 of these juveniles that survived until the following breeding season returned to Arizona. The individual that did not return showed evidence of permanent emigration to northern California. Our results contribute to a growing understanding of avian migration routes within the Pacific Flyway and the diverse migration strategies, including northward migration following the breeding season, exhibited across raptor populations. We highlight that explicit studies of juveniles, subadults, and nonbreeding adults are key to understanding avian migration dynamics and identifying opportunities for conservation.

Arizona↗

USGS2015

Who invented the mule deer ( Odocoileus hemionus )? On the authorship of the fraudulent 1812 journal of Charles Le Raye

The captivity journal of Charles Le Raye was first published in 1812 as a chapter in A topographical description of the state of Ohio, Indiana Territory, and Louisiana , a volume authored anonymously by a late officer in the U. S. Army. Le Raye was purported to be a French Canadian fur trader who, as a captive of the Sioux, had travelled across broad portions of the Missouri and Yellowstone river drainages a few years before the Lewis and Clark expedition (1804-1806), and his account of the land, its people, and its natural resources was relied upon as a primary source by generations of natural historians, geographers, and ethnographers. Based directly on descriptions of animals in the published journal, the naturalist Constantine S. Rafinesque named seven new species of North American mammals, including what are currently recognized as the mule deer ( Odocoileus hemionus ) and a Great Plains subspecies of white-tailed deer ( O. virginianus macrourus ). Unfortunately, Le Raye never existed, and historical, geographical, and ethnographical evidence indicates that the journal is fraudulent. Determining the author of this work is relevant to identifying the sources used to construct it, which may help us to understand the real animals upon which Rafinesque's species are based. Traditionally, authorship of the volume was attributed to Jervis Cutler, but his role in composing the fraudulent Le Raye journal has been called into question. In this paper, I present additional evidence supporting the hypothesis that Jervis Cutler bears primary responsibility for the Le Raye journal and that he had the background, opportunity, and potential motive to author it.

Archives of Natural History↗