Zika virus infection in the developing mouse produces dramatically different neuropathology dependent on viral strain

Zika virus (ZIKV) infection during pregnancy has been causally linked to a constellation of neurodevelopmental deformities in the fetus resulting in a disease termed congenital Zika syndrome (CZS). Here we detail how ZIKV infection produces extensive neuropathology in the developing mouse brain and spinal cord of both sexes. Surprisingly, neuropathology differs depending on viral strain with a French Polynesian isolate producing primarily excitotoxicity and a Brazilian isolate being almost exclusively apoptotic but occurring over a prolonged period that is more likely to produce severe hypoplasia. We also show exposure can produce a characteristic pattern of infection that mirrors neuropathology and ultimately results in gross morphological deformities strikingly similar to CZS. This research provides a valuable mouse model mirroring the clinical course of disease that can be used to test potential therapies to improve treatment and gain a better understanding of the disabilities associated with CZS

Seawater carbonate chemistry and survival and growth of young‑of‑the‑year golden king crab (Lithodes aequispinus)

Ocean acidification, a reduction in the pH of the oceans caused by increasing CO2, can have negative physiological effects on marine species. In this study, we examined how CO2-driven acidification affected the growth and survival of juvenile golden king crab (Lithodes aequispinus), an important fishery species in Alaska. Juveniles were reared from larvae in surface ambient pH seawater at the Kodiak Laboratory. Newly molted early benthic instar crabs were randomly assigned to one of three pH treatments: (1) surface ambient pH  8.2, (2) likely in situ ambient pH 7.8, and (3) pH 7.5. Thirty crabs were held in individual cells in each treatment for 127 days and checked daily for molting or death. Molts and dead crabs were photographed under a microscope and measured using image analysis to assess growth and morphology. Mortality was primarily associated with molting in all treatments, differed among all treatments, and was highest at pH 7.5 and lowest at ambient pH. Crabs at pH 7.5 were smaller than crabs at ambient pH at the end of the experiment, both in terms of carapace length and wet mass; had a smaller growth increment after molting; had a longer intermolt period. Carapace morphology was not affected by pH treatment. Decreased growth and increased mortality in laboratory experiments suggest that lower pH could affect golden king crab stocks and fisheries. Future work should examine if larval rearing conditions affect the juvenile response to low pH

Effects of ocean acidification on juvenile red king crab (Paralithodes camtschaticus) and Tanner crab (Chionoecetes bairdi) growth, condition, calcification, and survival.

Ocean acidification, a decrease in the pH in marine waters associated with rising atmospheric CO2 levels, is a serious threat to marine ecosystems. In this paper, we determine the effects of long-term exposure to near-future levels of ocean acidification on the growth, condition, calcification, and survival of juvenile red king crabs, Paralithodes camtschaticus, and Tanner crabs, Chionoecetes bairdi. Juveniles were reared in individual containers for nearly 200 days in flowing control (pH 8.0), pH 7.8, and pH 7.5 seawater at ambient temperatures (range 4.4-11.9 °C). In both species, survival decreased with pH, with 100% mortality of red king crabs occurring after 95 days in pH 7.5 water. Though the morphology of neither species was affected by acidification, both species grew slower in acidified water. At the end of the experiment, calcium concentration was measured in each crab and the dry mass and condition index of each crab were determined. Ocean acidification did not affect the calcium content of red king crab but did decrease the condition index, while it had the opposite effect on Tanner crabs, decreasing calcium content but leaving the condition index unchanged. This suggests that red king crab may be able to maintain calcification rates, but at a high energetic cost. The decrease in survival and growth of each species is likely to have a serious negative effect on their populations in the absence of evolutionary adaptation or acclimatization over the coming decades

Seawater carbonate chemistry and hemocyte physiology in the tanner crab (Chionoecetes bairdi)

We used flow cytometry to determine if there would be a difference in hematology, selected immune functions, and hemocyte pH (pHi), under two different, future ocean acidification scenarios (pH = 7.50, 7.80) compared to current conditions (pH = 8.09) for Chionoecetes bairdi, Tanner crab. Hemocytes were analyzed after adult Tanner crabs were held for two years under continuous exposure to acidified ocean water. Total counts of hemocytes did not vary among control and experimental treatments; however, there were significantly greater number of dead, circulating hemocytes in crabs held at the lowest pH treatment. Phagocytosis of fluorescent microbeads by hemocytes was greatest at the lowest pH treatment. These results suggest that hemocytes were dying, likely by apoptosis, at a rate faster than upregulated phagocytosis was able to remove moribund cells from circulation at the lowest pH. Crab hemolymph pH (pHe) averaged 8.09 and did not vary among pH treatments. There was no significant difference in internal pH (pHi) within hyalinocytes among pH treatments and the mean pHi (7.26) was lower than the mean pHe. In contrast, there were significant differences among treatments in pHi of the semi-granular+granular cells. Control crabs had the highest mean semi-granular+granular pHi compared to the lowest pH treatment. As physiological hemocyte functions changed from ambient conditions, interactions with the number of eggs in the second clutch, percentage of viable eggs, and calcium concentration in the adult crab shell was observed. This suggested that the energetic costs of responding to ocean acidification and maintaining defense mechanisms in Tanner crab may divert energy from other physiological processes, such as reproduction

Ocean Acidification Affects Hemocyte Physiology in the Tanner Crab (Chionoecetes bairdi).

We used flow cytometry to determine if there would be a difference in hematology, selected immune functions, and hemocyte pH (pHi), under two different, future ocean acidification scenarios (pH = 7.50, 7.80) compared to current conditions (pH = 8.09) for Chionoecetes bairdi, Tanner crab. Hemocytes were analyzed after adult Tanner crabs were held for two years under continuous exposure to acidified ocean water. Total counts of hemocytes did not vary among control and experimental treatments; however, there were significantly greater number of dead, circulating hemocytes in crabs held at the lowest pH treatment. Phagocytosis of fluorescent microbeads by hemocytes was greatest at the lowest pH treatment. These results suggest that hemocytes were dying, likely by apoptosis, at a rate faster than upregulated phagocytosis was able to remove moribund cells from circulation at the lowest pH. Crab hemolymph pH (pHe) averaged 8.09 and did not vary among pH treatments. There was no significant difference in internal pH (pHi) within hyalinocytes among pH treatments and the mean pHi (7.26) was lower than the mean pHe. In contrast, there were significant differences among treatments in pHi of the semi-granular+granular cells. Control crabs had the highest mean semi-granular+granular pHi compared to the lowest pH treatment. As physiological hemocyte functions changed from ambient conditions, interactions with the number of eggs in the second clutch, percentage of viable eggs, and calcium concentration in the adult crab shell was observed. This suggested that the energetic costs of responding to ocean acidification and maintaining defense mechanisms in Tanner crab may divert energy from other physiological processes, such as reproduction

Seawater carbonate chemistry and micromechanical properties of the mineralized cuticle in juvenile red and blue king crabs

Ocean acidification (OA) adversely affects a broad range of marine calcifying organisms. Crustaceans, however, exhibit mixed responses to OA, with growth or survival negatively affected in some species, but unaffected or positively affected in others. In crustaceans, the mineralized cuticle resists mechanical loads, provides protection from the environment, and enables mobility, but little is known about how OA or interactions between OA and temperature affect its structure or function. Here, the effects of OA on the mechanics, structure, and composition of the cuticle in two Alaska king crab species was assessed. Juvenile blue king crabs (Paralithodes platypus) were exposed for a year to three pH levels, 8.1 (ambient), 7.8 and 7.5. Juvenile red king crabs (Paralithodes camtschaticus) were exposed for ~ 6 months to two pH levels, 8.0 and 7.8, at three temperatures: ambient, ambient + 2 °C, and ambient + 4 °C. Cuticle microhardness (a measure of resistance to permanent or plastic mechanical deformation), thickness, ultrastructure, and elemental composition were assessed in two body regions, the carapace and the crushing chela (claw). In both species tested, OA reduced endocuticle microhardness in the chela, but not in the carapace. There was no effect of pH or temperature on total procuticle thickness of the chela or carapace in either species. Reductions in microhardness were not driven by reduced calcium content of the shell. In fact, calcium content was significantly elevated in the carapace of blue king crabs and in the chela of red king crabs exposed to lower than ambient pH at ambient temperature, suggesting that calcium content alone is not a sufficient proxy for mechanical properties. Reduced chela microhardness, indicative of more compliant material, could compromise the utility of crushing chelae in feeding and defense

Condition index and calcification in crabs held in Control and Acidified water.

<p>Condition index of red king crabs (A) and Tanner crabs (B) and percent calcium (dry mass) in red king crabs (C) and Tanner crabs (D) at the end of the experiment. Bars are mean+SE. Bars with different letters above them differ significantly.</p

The mean and standard error (SE) of water chemistry parameters measured (DIC, and Alkalinity) and calculated (all others, see text for details) in the three treatments during the experiments.

<p>N = 23–28 per treatment for DIC and Alkalinity and N = 200 per treatment for pH_F.</p

Survival of crabs in Control and Acidified water.

<p>Red king crab (A), and Tanner crabs (B) in control and treatment tanks over the duration of the experiment. Maximum likelihood estimated mortality rates±standard error were Control −0.0023±0.00007 day⁻¹, pH 7.8−0.0047±0.00011 day⁻¹, pH 7.5−0.025±0.00066 day⁻¹ for red king crab and Control -0.0010±0.00004 day⁻¹, pH 7.8-0.0023±0.00007 day⁻¹, pH 7.5-0.0050±0.00011 day⁻¹ for Tanner crab. See text for model details.</p

Changes in crab morphology in Control and Acidified water.

<p>Mean PC1 scores (±SE) from principle component analysis of red king crab (A) and Tanner crab (B) morphometrics for each molt for crabs held in Control, pH 7.8, and pH 7.5 water. Symbols with different letters beside them differ significantly within that molt stage. Note that crab size is inversely proportional to PC1 for both species.</p