Oxidative stress and histopathological effects by microplastic beads, in the crayfish Procambarus clarkii, and fiddler crab Leptuca pugilator.

The present study was aimed at evaluating the in vivo effects of microplastics (MP), in terms of oxidative stress and histopathological effects, in two crustacean species: Procambarus clarkii and Leptuca pugilator. In addition, MP accumulation in the hepatopancreas (HP) of both species was also determined. Adults of both crayfish and crabs were exposed for one month to fluorescent polystyrene beads (size: 1 µm) at nominal concentrations of 1000 or 5000 particles/mL. During the exposure, animals were maintained under controlled feeding, aeration, temperature, and photoperiod conditions. At the end of the exposure, HP and hemolymph (HL) samples were harvested for analysis of oxidative damage and total antioxidant levels. Additionally, the presence of MPs in both tissues was confirmed. Significant differences with the control groups were observed in lipid peroxidation levels in HP in animals exposed to the lowest concentration in P. clarkii and to the highest concentration in L. pugilator. A marked increase in antioxidant levels was also observed in the HL at both concentrations in P. clarkii, and at the highest MPs concentration in L. pugilator. Moreover, several histopathological changes were detected in both gills and HP, including hypertrophied lamellae, lifting or collapse of gill epithelia, loss of normal shape of hepatopancreatic tubules, and epithelial atrophy in the HP tissue. We conclude that exposure to MP beads at selected concentrations results in oxidative damage, induces histopathological changes in gills and HP, and triggers an antioxidant response in two crustacean species.

Ontogeny of fetal hepatic and placental growth and metabolism in sheep.

Ontogeny of fetal hepatic and placental growth and in vitro oxygen consumption (VO2) was investigated in fetal lambs at 75, 100, and 136 days postconception. Fetal hepatic relative weight and placental absolute and relative weights declined during this period. Oxygen consumption per gram dry weight of fetal liver and maternal placenta declined between mid and late gestation while fetal placental VO2 was unchanged. Estimated VO2 of the whole placenta did not change while the estimated total hepatic VO2 increased more than threefold between 75 and 136 days. Total hepatic VO2 was highly correlated with total placental VO2 at 136 days (r = 0.84). The results suggest that the placenta reaches its maximum growth and metabolic capacity before 100 days and possibly at or before midgestation. Changes in hepatic weight-specific total VO2, in addition to the declining relative size of the fetal liver, must contribute to the progressive decline in metabolic rate of the whole fetus during the second half of pregnancy. Correlations between placental and fetal liver weights and metabolic rates suggest the possibility of placental regulation of fetal hepatic growth and metabolism.

Effect of heat stress on ovine placental growth in early pregnancy.

Ditocous Dorset ewes were fed to predicted requirements and kept in environmental chambers at 21 degrees C (n = 6) or 40 degrees C (n = 5) between days 50 and 75 of gestation. Ewes were slaughtered and the pregnant uterus was dissected for measurement of conceptus weights and in vitro estimations of placental mitotic activity. Heat caused a 19% reduction in placental weight but did not affect fetal weight. Placental DNA and protein concentrations and protein/DNA were similar in both groups. Total placental DNA content was significantly reduced in the heated ewes, suggesting a reduction in cell number; however, DNA synthetic rate tended to be higher. These results are consistent with the hypothesis that fetal growth retardation in chronically heat-stressed ewes occurs in late pregnancy as a consequence of a primary reduction in placental growth in early gestation.

Chronic heat stress and prenatal development in sheep: II. Placental cellularity and metabolism.

Aspects of placental protein and energy metabolism were examined in pregnant ewes subjected to either thermoneutral (TN, 18 to 20 degrees C, 30% humidity, n = 7) or hot (H, 30 to 40 degrees C, 40% humidity, n = 5) temperatures through mid and late gestation. Fetal and placental weights and total content of protein, RNA, and DNA were reduced (P less than .001) in H ewes. Placental protein and RNA concentrations (mg/g) were not different, and DNA concentrations were slightly greater (P less than .1), in H vs TN ewes. Thus, heat seemed to greatly reduce total cell number and placentome size and only slightly decrease cell size. Ratios of RNA to DNA indicated a reduced capacity for protein synthesis in H placenta. However, in vitro fractional rates of protein synthesis in tissue slices from the fetal and maternal placenta and from the myoendometrium were not different between TN and H ewes. The H ewes had greater placental protein concentrations of hydroxyproline and glycine, perhaps suggesting a greater collagen content. In vitro oxygen consumption of fetal placenta, but not of maternal placenta or myoendometrium, was lower in H than in TN ewes. This lower oxygen consumption was partially due to a lower Na+,K+ ATPase-dependent oxygen consumption.

Growth and metabolism of the placenta after unilateral fetectomy in twin pregnant ewes.

Twin-pregnant ewes underwent unilateral fetectomy (Fetx) at 50 days of gestation and were studied at 136 days. Aspects of conceptus growth and placental cellularity and metabolism in vitro were compared to those of unoperated control groups of twin-pregnant or single-pregnant ewes. Mean fetal weight in Fetx ewes tended to be greater than that of twin-pregnant ewes and was similar to that of single-pregnant ewes. Mean placental wet and dry weights were intermediate between those for naturally single- and twin-pregnant animals. Fetectomy caused a significant increase in placental protein:DNA ratio but an unchanged DNA concentration, apparently due to cellular hypertrophy in the placenta of the remaining fetus. Weight-specific rate of oxygen consumption (VO2) of fetal placental tissue in twin-pregnant ewes was higher than in Fetx or singles while maternal placental VO2 in twins tended to be lower than in either of the other two groups. These results highlight the plasticity of placental metabolism and growth, perhaps in response to altered trophic signals from the fetus. Unilateral fetectomy should prove useful in studies designed to identify these signals.

Regression of brown adipose tissue mitochondrial function and structure in neonatal goats.

Rapidly regressing perirenal brown adipose tissue (BAT) of neonatal goats was studied to correlate changes in mitochondrial metabolism and thermogenic capacity with changes in mitochondrial structure. The alpha-glycerophosphate shuttle activity of perirenal BAT mitochondria declined 60% from birth to 6 days of age. Oxygen consumption and thermogenic capacity measured by ion conductance peaked at birth and declined to low levels at 6 days. Sample electron micrographs of perirenal BAT showed intact electron-dense mitochondria with many cristae and little matrix area at 2 days. However, by 6 days the mitochondria were very relaxed with large matrix area, few cristae, and observable degradation. These results indicate that the morphological changes exhibited by rapidly regressing goat's perirenal BAT in the 1st wk postpartum are accompanied by dramatic alterations in BAT mitochondrial function.