Vitamin C is an essential water-soluble nutrient which cannot be synthesised or stored by humans. It is a potent antioxidant with anti-inflammatory and immune-supportive roles. Previous research has indicated that vitamin C levels are depleted in critically ill patients. In this study we have assessed plasma vitamin C concentrations in critically ill patients relative to infection status (septic shock or non-septic) and level of inflammation (C-reactive protein concentrations). Vitamin C status was also assessed relative to daily enteral and parenteral intakes to determine if standard intensive care unit (ICU) nutritional support is adequate to meet the vitamin C needs of critically ill patients. Forty-four critically ill patients (24 with septic shock, 17 non-septic, 3 uncategorised) were recruited from the Christchurch Hospital Intensive Care Unit. We measured concentrations of plasma vitamin C and a pro-inflammatory biomarker (C-reactive protein) daily over 4 days and calculated patients’ daily vitamin C intake from the enteral or total parenteral nutrition they received. We compared plasma vitamin C and C-reactive protein concentrations between septic shock and non-septic patients over 4 days using a mixed effects statistical model, and we compared the vitamin C status of the critically ill patients with known vitamin C bioavailability data using a four-parameter log-logistic response model. Overall, the critically ill patients exhibited hypovitaminosis C (i.e., < 23 μmol/L), with a mean plasma vitamin C concentration of 17.8 ± 8.7 μmol/L; of these, one-third had vitamin C deficiency (i.e., < 11 μmol/L). Patients with hypovitaminosis C had elevated inflammation (C-reactive protein levels; P < 0.05). The patients with septic shock had lower vitamin C concentrations and higher C-reactive protein concentrations than the non-septic patients (P < 0.05). Nearly 40% of the septic shock patients were deficient in vitamin C, compared with 25% of the non-septic patients. These low vitamin C levels were apparent despite receiving recommended intakes via enteral and/or parenteral nutritional therapy (mean 125 mg/d). Critically ill patients have low vitamin C concentrations despite receiving standard ICU nutrition. Septic shock patients have significantly depleted vitamin C levels compared with non-septic patients, likely resulting from increased metabolism due to the enhanced inflammatory response observed in septic shock.

Female mate choice after copulation or spawning is cryptic when a female differentially influences the fertilization success of sperm from different males. We tested whether ovarian fluid could act as a potential mechanism of cryptic female choice (CFC) in chinook salmon (Oncorhynchus tshawytscha) by comparing how ovarian fluid from each of 7 females affected the sperm behavior of 11 different males. Using computer-assisted sperm analysis, we measured sperm velocity, motility, longevity, and linearity from the ejaculates of each male activated in the ovarian fluid from each female. Mean sperm swimming speed, path trajectory, and longevity differed significantly among males, and within males depended on the female's ovarian fluid in which it was activated. Most important, the pattern of within-male variation in these traits also varied significantly among males in response to different females’ ovarian fluids. As sperm velocity is known to be a prime determinant of fertilization success in externally fertilizing fishes. This finding suggests that variation in the composition of female ovarian fluid may be a mechanism for CFC, whereby females differentially enhance the swimming speed of sperm from different males. Thus, female ovarian fluid can alter relative male fertilization success when there is intense sperm competition, as there is in this and other group spawning fish species.

In this study, we investigated two potentially important intersexual postcopulatory gametic interactions in a population of chinook salmon ( Oncorhynchus tshawytscha ): (i) the effect of female ovarian fluid (OF) on the behaviour of spermatozoa during fertilization and (ii) the effects of multilocus heterozygosity (MLH) (as an index of male quality) and female–male genetic relatedness on sperm behaviour and male fertilization success when there is sperm competition in the presence of that OF. To do this, we conducted a series of in vitro competitive fertilization experiments and found that, when ejaculates from two males are competing for access to a single female's unfertilized eggs, fertilization success was significantly biased towards the male whose sperm swam fastest in the female's OF. Embryo survival—a measure of fitness—was also positively correlated with both sperm swimming speed in OF and male MLH, providing novel evidence that cryptic female choice is adaptive for the female, enhancing the early survival of her offspring and potentially influencing her fitness.

Disentangling the relative roles of males, females and their interactive effects on competitive fertilization success remains a challenge in sperm competition. In this study, we apply a novel experimental framework to an ideally suited externally fertilizing model system in order to delineate these roles. We focus on the chinook salmon,Oncorhynchus tshawytscha, a species in which ovarian fluid (OF) has been implicated as a potential arbiter of cryptic female choice for genetically compatible mates. We evaluated this predicted sexually selected function of OF using a series of factorial competitive fertilization trials. Our design involved a series of 10 factorial crosses, each involving two ‘focal’ rival males whose sperm competed against those from a single ‘standardized’ (non-focal) rival for a genetically uniform set of eggs in the presence of OF from two focal females. This design enabled us to attribute variation in competitive fertilization success among focal males, females (OF) and their interacting effects, while controlling for variation attributable to differences in the sperm competitive ability of rival males, and male-by-female genotypic interactions. Using this experimental framework, we found that variation in sperm competitiveness could be attributed exclusively to differences in the sperm competitive ability of focal males, a conclusion supported by subsequent analyses revealing that variation in sperm swimming velocity predicts paternity success. Together, these findings provide evidence that variation in paternity success can be attributed to intrinsic differences in the sperm competitive ability of rival males, and reveal that sperm swimming velocity is a key target of sexual selection.

Sperm traits of externally fertilizing fish species are typically measured in fresh (or salt) water, even though the spawning environment of their ova contains ovarian fluid. In this study, we measured sperm traits of Chinook salmon ( Oncorhynchus tshawytscha (Walbaum in Artedi, 1792)) in both fresh water and dilute ovarian fluid at 10 and 20 s postactivation, using a computer-assisted sperm analysis system. Spermatozoa swam faster, and had both higher percent motility and a straighter path trajectory for a longer period of forward motility when activated in ovarian fluid compared with activation in fresh water. Comparing sperm activity of 10 males in water versus ovarian fluid, we found a weak but significant correlation for sperm swimming speed at 10 s postactivation (r = 0.34, p = 0.01), but not for any other sperm traits measured. Most important, across males, mean sperm swimming speed in water accounted for <10% of the observed variation in mean sperm swimming speed in ovarian fluid. Thus, we argue that sperm traits measured in fresh water are not particularly relevant to those same traits during normal spawning in this species. We suggest that sperm performance measured in fresh water should be used with caution when comparing the potential for individual males to fertilize ova, especially in studies of sperm competition in externally fertilizing species.