Karen McBee

In situ (mesocosm) assessment of immunotoxicity risks to small mammals inhabiting petrochemical waste sites

Oil refineries inadvertently deposit a variety of complex mixtures of organic hydrocarbons and heavy metals in the soil, many of which are thought to be potent immunotoxicants. Terrestrial ecosystems such as this have not been adequately investigated with respect to wild rodent populations. The primary objective of this study was to use mesocosms to assess the immunotoxicity risks to feral small mammal populations associated with soils contaminated with petroleum refinery wastes. A series of 4-week and 8-week exposure trials using laboratory raised cotton rats (Sigmodon hispidus) were conducted in situ on three contaminated and three reference sites on the Oklahoma Refining Company Superfund Waste Site, Cyril, Oklahoma. Cotton rats exposed to these soils showed significant alterations in selected morphological traits, in vivo humoral immune responses, complement activity, and macrophage activity. However, immune alterations were not great, suggesting that resident small mammals may be a better biomonitoring choice than using mesocosms.

Induction of hepatic CYP1A activity as a biomarker for environmental exposure to Aroclor® 1254 in feral rodents

Specimens of the feral mouse species Reithrodontomys fulvescens trapped from a polychlorinated biphenyl (PCB)-contaminated field location had hepatic ethoxy-resorufin (ETR) O-dealkylase activities and immunoreactive CYP1A protein contents which were two- to threefold higher than those measured in animals of the same species and sex collected from non PCB-contaminated reference sites. Specimens with hepatic ETR O-dealkylase activities differing by as little as 50% could readily be assigned as originating from the PCB or reference sites by the use of a specific chemical inhibitor of cytochrome P450IA (CYP1A). The relative levels of ETR O-dealkylase activity in R. fulvescens significantly correlated with hepatic PCB burdens (r=0.819, P<0.01). When the magnitudes of the induced ETR O-dealkylase activities corresponding to given hepatic PCB burdens were compared between the feral animals, F344/NCr rats (Rattus norvegicus) or B6C3F1 mice (Mus musculus) exposed in the laboratory to dietary Aroclor® 1254, the order of sensitivity to the inducing effects of PCBs were F344/NCr rat>B6C3F1 mouse>R. fulvescens.

Allelic diversity at the Mhc-DQA locus in cotton rats (Sigmodon hispidus) and a comparison of DQA sequences within the family muridae (Mammalia: Rodentia).

Abstract The cotton rat (Sigmodon hispidus) is a common murid rodent of the southern United States, Mexico, and Central America. Using single-stranded conformation polymorphism analysis and DNA sequencing techniques, 11 DQA exon 2 alleles were detected among 180 S. hispidus from Caddo County, Oklahoma, USA. The alleles represent a single locus exhibiting a high level of polymorphism. Nucleotide and amino acid distance values among DQA alleles of S. hispidus were higher than those within Mus musculus and species of Rattus. Although the distribution of polymorphic amino acid residues among alleles of S. hispidus was similiar to that of Mus and Rattus, some residues of the α-helix region were more variable in S. hispidus. Comparisons of nonsynonymous and synonymous substitutions indicated a trend toward higher numbers of nonsynonymous substitutions; however, this difference was not significant statistically among S. hispidus alleles. To examine evolution of DQA alleleswithin Muridae, we performed a phylogenetic analysis that included DQA alleles from S. hispidus, Peromyscus leucopus, M. musculus, R. norvegicus, and six Australian species of Rattus. Results depicted monophyly for each genus, and this concordance between species and gene trees represents a lack of evidence for trans-species persistence of alleles among these genera.

Fluorosis in a wild cotton rat (Sigmodon hispidus) population inhabiting a petrochemical waste site

We have developed an in situ mammalian model for evaluating environmental contamination using wild cotton rats. In a series of experiments, 200 male cotton rats were captured during 4 collection periods (spring 1991 = 35; fall 1991 = 60; spring 1992 = 53; fall 1992 = 52). A total of 103 of these cotton rats were captured from control sites, and the remaining 97 were captured from an abandoned oil refinery. All sites were located in the vicinity of Cyril, Oklahoma. There were alterations in the incisors of cotton rats captured from the refinery site. Normal color of cotton rat incisors is deep yellow-orange, which is imparted by a pigment normally produced by ameloblasts. Grossly, the upper incisors of 37 of 97 rats and lower incisors of 54 of 97 rats were affected. The affected incisors were white, chalky, and thin with striations and erosions of the enamel. Microscopic examination revealed that there were dysplastic and necrotic changes in the ameloblasts. The bone fluoride levels were significantly higher in rats captured from the refinery as compared to the rats captured from the control sites.

Cross-placental transfer of organochlorine pesticides in mexican free-tailed bats from Oklahoma and New Mexico

A total of 41 pregnant Mexican free-tailed bats (Tadarida brasiliensis) were collected from Carlsbad Caverns, New Mexico, and Vickery Cave, Oklahoma, in May and June 1990 and May 1991 for organochlorine pesticide analyses. Residues of p,p′-DDE were detected by gas chromatography in 40 embryos at levels highly correlated with brain concentrations (r=0.496) but were not correlated mothers body tissues (r=0.060). Embryonic levels also did not correlate with either embryonic fat content (r=0.018) or maternal fat content (r=0.300), suggesting that placental membranes offer only marginal protection for developing embryos against exposure to lipophilic organochlorine pesticides.

Karyotypic Variation in the Genus Platemys (Testudines: Pleurodira)

Standard karyotypes are reported for all currently recognized members of the South American genus Platemys. Platemys platycephala (2n = 64) has a karyotype distinct from the other four members of the genus (P. macrocephala, 2n = 48; P. pallidipectoris, P. radiolata, P. spixii 2n = 50). The range in diploid numbers within this genus is greater than all other members of the suborder Pleurodira. Such karyotypic variability is particularly uncommon among closely related turtles and supports generic separation of P. platycephala from the remaining four species. These four have karyotypes similar to Australian chelids, and, with Chelus, may represent a group intermediate between Australian and the other South American forms. THE two families of living pleurodiran tur

Population genetics of the hispid cotton rat (Sigmodon hispidus): patterns of genetic diversity at the major histocompatibility complex

The hispid cotton rat, Sigmodon hispidus, is a common rodent widely distributed across the southern USA and south into South America. To characterize major histocompatibility complex (MHC) diversity in this species and to elucidate large‐scale patterns of genetic partitioning, we examined MHC genetic variability within and among 13 localities, including a disjunct population in Arizona and a population from Costa Rica that may represent an undescribed species. We also tested the hypothesis that populations within the USA are at equilibrium with regard to gene flow and genetic drift, resulting in isolation‐by‐distance. Using single‐strand conformation polymorphism (SSCP) analysis we identified 25 alleles from 246 individuals. Gene diversity within populations ranged from 0.000 to 0.908. Analysis of molecular variance (amova) revealed that 83.7% of observed variation was accounted for by within‐population diversity and 16.3% was accounted for by among‐population divergence. The disjunct population in Arizona was fixed for a single allele. The Costa Rican population was quite divergent based on allelic composition and was the only population with unique alleles. Within the main portion of the geographical distribution of S. hispidus in the USA there was considerable divergence among some populations; however, there was no significant pattern of isolation‐by‐distance overall (P = 0.090). Based on the significant divergence of the only sampled population to its east, the Mississippi River appears to represent a substantial barrier to gene flow.

Intraspecific phylogeography of red squirrels (Tamiasciurus hudsonicus) in the central Rocky Mountain region of North America.

We used variation in a portion of the mitochondrial DNA control region to examine phylogeography of Tamiasciurus hudsonicus, a boreal-adapted small mammal in the central Rocky Mountain region. AMOVA revealed that 65.66% of genetic diversity was attributable to variation within populations, 16.93% to variation among populations on different mountain ranges, and 17.41% to variation among populations within mountain ranges. Nested clade analysis revealed two major clades that likely diverged in allopatry during the Pleistocene: a southern clade from southern Colorado and a northern clade comprising northern Colorado, Wyoming, eastern Utah, and eastern Idaho. Historically restricted gene flow as a result of geographic barriers was indicated between populations on opposite sides of the Green River and Wyoming Basin and among populations in eastern Wyoming. In some instances genetic structure indicated isolation by distance.

Organochlorine pesticide accumulation and genotoxicity in Mexican free-tailed bats from Oklahoma and New Mexico

The summer population of Mexican free-tailed bats (Tadarida brasiliensis) at Carlsbad Caverns, New Mexico, declined from an estimated 8.7 million in 1936 to 200,000 in 1974; thereafter, it increased to approximately 700,000 in 1991. This decline has been attributed primarily to organochlorine (OC) pesticide contamination and habitat disturbance. Similar declines have been observed in other populations of this species. This study examined the potential genotoxic effects of OC pesticide contamination on two populations of T. brasiliensis. Pesticide accumulation, frequencies of chromosomal aberrancy, and nuclear DNA content variation in spleen and testicular tissues were examined in specimens collected from Carlsbad Caverns and Vickery Cave, a maternity colony in northwestern Oklahoma, during the summers of 1990 and 1991. Pesticide residues in brain and carcass tissues were identified and quantified by electron capture gas chromatography. Genotoxicity was examined with the standard bone marrow chromosomal aberration assay and flow cytometry. Statistical relationships among pesticide content, observed chromosomal aberrancy, and nuclear DNA content variation were examined. Both populations demonstrated significant levels of DDE contamination; however, the Carlsbad Caverns population showed consistently higher pesticide loads. Males also demonstrated higher levels than females. No statistical differences in chromosomal aberrancy or nuclear DNA content variation were observed among sexes, sites, or collected periods. Positive correlations were detected between brain and carcass DDE concentrations for all bats examined. A significant negative relationship was found between brain DDE concentration and coefficients of variation in spleen DNA content only for males.

FLUOROSIS RISKS TO RESIDENT HISPID COTTON RATS ON LAND- TREATMENT FACILITIES FOR PETROCHEMICAL WASTES

Land-treatment of petroleum wastes is a widely used industrial practice, yet there has been no comprehensive evaluation of the long-term risks to human or terrestrial ecosystems from such practices. We evaluated cotton rat (Sigmodon hispidus) populations on three sites in Oklahoma (USA) that historically used land-treatment for disposal of various petroleum wastes (July 1995–March 1997). Average concentrations of fluoride in soil from these sites ranged from 878 to 4317 mg/kg. A census of resident cotton rats on land-treatment sites revealed a high incidence (40% overall) of dental lesions compared to reference populations (<1% dental lesions). During winter there was a 34% to 65% increase compared to summer in frequency of dental lesions in cotton rats on two of the three land-treatment sites. Incidence of dental lesions on two land-treatment sites was greater (9–16%) in female cotton rats compared to males. Cotton rats from land-treatment sites had higher concentrations of fluoride in bone and greater severity of dental lesions compared to reference animals. Dental lesions were considered to be most consistent with dental fluorosis because of elevated fluoride in bone. Neither concentration of fluoride in soil nor level of fluoride in bone was a good predictor of severity of dental lesions in cotton rats on land-treatment sites.