The influence of incubation temperature on offspring traits varies across northern and southern populations of the American alligator (Alligator mississippiensis).

Maternal provisioning and the developmental environment are fundamental determinants of offspring traits, particularly in oviparous species. However, the extent to which embryonic responses to these factors differ across populations to drive phenotypic variation is not well understood. Here, we examine the contributions of maternal provisioning and incubation temperature to hatchling morphological and metabolic traits across four populations of the American alligator (Alligator mississippiensis), encompassing a large portion of the species' latitudinal range. Our results show that whereas the influence of egg mass is generally consistent across populations, responses to incubation temperature show population-level variation in several traits, including mass, head length, head width, and residual yolk mass. Additionally, the influence of incubation temperature on developmental rate is greater at northern populations, while the allocation of maternal resources toward fat body mass is greater at southern populations. Overall, our results suggest that responses to incubation temperature, relative to maternal provisioning, are a larger source of interpopulation phenotypic variation and may contribute to the local adaptation of populations.

Maternal provisioning interacts with incubation temperature to affect hatchling mercury exposure in an oviparous reptile.

The thermal environment experienced by developing embryos can influence the utilization of maternally provisioned resources. Despite being particularly consequential for oviparous ectotherms, these dynamics are largely unexplored within ecotoxicological frameworks. Here, we test if incubation temperature interacts with maternally transferred mercury to affect subsequent body burdens and tissue distributions of mercury in hatchling American alligators (Alligator mississippiensis). Nine clutches of alligator eggs were collected from a mercury-contaminated reservoir and incubated at either female- or male-promoting temperatures. Total mercury (THg) concentration was measured in egg yolk collected during incubation and in a suite of tissues collected from hatchlings. THg concentrations in residual yolk and blood were higher in hatchlings incubated at cooler, female-promoting temperatures compared to the warmer, male-promoting temperatures. THg concentrations in most tissues were positively correlated with THg concentrations in blood and dermis, and egg yolk THg concentration was the best predictor of THg concentration in many resultant tissues. Our results highlight a hereto unknown role of the developmental environment in mediating tissue specific uptake of contaminants in an oviparous reptile.

Host use by 2 sibling species of bogus yucca moths in relation to plant hardness and saponin content.

Plant defenses allow plants to deter or kill their insect herbivores and are considered to be a major driver of host use for herbivorous insects in both ecological and evolutionary time. Many closely related species of insect herbivores differ in their ability to respond to plant defenses and in some cases are specialized to specific plant species. Here we tested whether both mechanical and chemical plant defenses are a major factor in determining the host range of 2 sibling species of Prodoxid bogus yucca moths, Prodoxus decipiens (Riley) and Prodoxus quinquepunctellus (Chambers) that feed within the inflorescence stalk of Yucca species. These 2 moth species have separate suites of host plant species, yet narrowly overlap geographically and share 1 Yucca species, Y. glauca. We surveyed the lignin and cellulose content, the force required to the puncture the stalk tissue, and saponin concentration across 5 Yucca species used as hosts. Lignin, cellulose concentrations, and stalk hardness differed among Yucca species but did not correlate with host use patterns by the moths. Saponin concentrations in the stalk tissue were relatively low for yuccas (<1%) and did not differ among species. The results suggest that these moth species should be able to use each other's hosts for egg deposition. Additional factors such as larval development or competition among larvae for feeding space may serve to keep moth species from expanding onto plants used by its sibling species.

Land-use change is associated with multi-century loss of elephant ecosystems in Asia.

Understanding historic patterns of land use and land cover change across large temporal and spatial scales is critical for developing effective biodiversity conservation management and policy. We quantify the extent and fragmentation of suitable habitat across the continental range of Asian elephants (Elephas maximus) based on present-day occurrence data and land-use variables between 850 and 2015 A.D. We found that following centuries of relative stability, over 64% (3.36 million km2) of suitable elephant habitat across Asia was lost since the year 1700, coincident with colonial-era land-use practices in South Asia and subsequent agricultural intensification in Southeast Asia. Average patch size dropped 83% from approximately 99,000-16,000 km2 and the area occupied by the largest patch decreased 83% from ~ 4 million km2 (45% of area) to 54,000 km2 (~ 7.5% of area). Whereas 100% of the area within 100 km of the current elephant range could have been considered suitable habitat in the year 1700, over half was unsuitable by 2015, driving potential conflict with people. These losses reflect long-term decline of non-forested ecosystems, exceeding estimates of deforestation within this century. Societies must consider ecological histories in addition to proximate threats to develop more just and sustainable land-use and conservation strategies.

Relationships between maternally-transferred mercury and hatchling development, behavior, and survival in the American alligator (Alligator mississippiensis).

Mercury is a toxic and pervasive environmental contaminant that can be transferred from mother to offspring during development. Consequences of maternally-transferred mercury have been observed in vertebrate taxa, including reduced clutch viability, reduced offspring size, and behavioral alterations. These sublethal effects have been assumed to decrease survivorship, though this is seldom assessed. Here, we examined how maternally-transferred mercury interacts with incubation temperature to influence reproductive success, offspring behavior, and subsequent survival in the American alligator (Alligator mississippiensis). We collected nine clutches of eggs from a mercury contaminated reservoir on the Savannah River Site, South Carolina, and incubated eggs at either female- or male-promoting temperatures. Clutch-averaged mercury in egg yolk was high relative to other studies in crocodilians and ranged from 0.248 to 0.554 ppm compared to 0.018-0.052 ppm at a site with low levels of mercury contamination; mercury levels in hatchling blood ranged from 0.090 to 0.490 ppm (x¯ = 0.240 ppm, n = 158). We found few, mostly negligible correlations between life history traits and mercury but noted a positive relationship with egg mass, possibly mediated by correlated maternal effects such as resource provisioning. Incubation temperature exerted strong effects on hatchling phenotypes, with warmer, male-promoting temperatures producing larger and bolder hatchlings. Presumptive females, produced from cooler incubation temperatures, spent more time in warm areas during behavior trials. Hatchlings were released 10-15 days post-hatch and surveyed over eight months to assess survival. Survivorship was positively correlated with hatchling size and negatively correlated with proportional time spent in warm areas. Presumptive females had much lower survival, and overall survivorship for the eight-month period was 0.185-0.208, depending on the modelling approach. Our study suggests that, within the range of concentrations we observed, incubation temperature has a stronger effect on offspring behavior and survival than maternally-transferred mercury pollution in American alligators.

Sex Determination and Ovarian Development in Reptiles and Amphibians: From Genetic Pathways to Environmental Influences.

BACKGROUND: Reptiles and amphibians provide untapped potential for discovering how a diversity of genetic pathways and environmental conditions are incorporated into developmental processes that can lead to similar functional outcomes. These groups display a multitude of reproductive strategies, and whereas many attributes are conserved within groups and even across vertebrates, several aspects of sexual development show considerable variation. SUMMARY: In this review, we focus our attention on the development of the reptilian and amphibian ovary. First, we review and describe the events leading to ovarian development, including sex determination and ovarian maturation, through a comparative lens. We then describe how these events are influenced by environmental factors, focusing on temperature and exposure to anthropogenic chemicals. Lastly, we identify critical knowledge gaps and future research directions that will be crucial to moving forward in our understanding of ovarian development and the influences of the environment in reptiles and amphibians. KEY MESSAGES: Reptiles and amphibians provide excellent models for understanding the diversity of sex determination strategies and reproductive development. However, a greater understanding of the basic biology of these systems is necessary for deciphering the adaptive and potentially disruptive implications of embryo-by-environment interactions in a rapidly changing world.

Implementing a Hybrid Expression Method That Allows Upper-Division Biochemistry Lab Students To Engage in a Full Protein Production Experience While Allowing Ample Time for Characterization Experiments.

Protein structure, function, and signaling are a large portion of biochemistry. Because of this, proteins are often used as model systems in biochemistry laboratory courses, where a course-long project might comprise protein expression, purification, and characterization. Two common protein expression methods are isopropyl ß-d-1-thiogalactopyranoside (IPTG) induction, which utilizes easy-to-make media but requires extensive cell-growth monitoring that is time-intensive, and autoinduction, which employs multicomponent media that are time-consuming to make but require no cell-growth monitoring. A protein expression method that is a hybrid of IPTG induction and autoinduction is presented. The hybrid method utilizes the medium of IPTG induction and the no-cell-growth-monitoring induction process of autoinduction, saving hands-on time in the protein expression phase to allow more time for protein characterization while still having students execute each step.

Improved Synthesis of 2-Trifluoromethyl-10-aminopropylphenothiazine: Making 2-Trifluoromethyl-10-aminopropylphenothiazine Readily Available for Calmodulin Purification.

An improved and high yielding three-step synthesis for the production of 2-trifluoromethyl-10-aminopropylphenothiazine (TAPP) using less hazardous and more inexpensive reagents, its coupling to Sepharose-4B resin, and its ability to purify calmodulin are described. The overall yield of TAPP, starting with 3-aminopropyl bromide hydrobromide and 2-(trifluoromethyl)phenothiazine, was 96%.