Diet Affects the Temperature-Size Relationship in the Blowfly Aldrichina grahami.

In warmer environments, most ectotherms exhibit a plastic reduction in body size (the temperature-size rule, TSR). However, in such environments, growth is usually accelerated and would be expected to result in maturation at a larger body size, leading to increases in fecundity, survival, and mating success, compared to maturation at a smaller size (the 'life-history puzzle'). To explore these mechanisms, we reared Aldrichina grahami at 20 °C, 25 °C, and 30 °C, and added a nutritional challenge by using dilutions of pork liver paste to provide diets that ranged in quality from high (undiluted) to moderate (1/8), low (1/16), and poor (1/24). Larvae were randomly sampled for weighing from hatching. Growth curves were fitted to the relationships between growth rate and weight for the third instar larvae. Our results showed that body size was affected by an interaction between temperature and diet, and that following or not following the TSR can vary depending on underfeeding. Moreover, when the TSR was followed as temperature increased, there was a cross-over point that divided the two growth curves into early and later stages, which could be used to help understand the life-history puzzle in warmer temperatures, with the instantaneous growth rate being faster in the early stages of development and then slower in later stages. This study reminds us that animals have evolved to cope with multiple simultaneous environmental changes, and it has thus offered a better understanding of life-history puzzles.

Effects of Diet on the Gut Bacterial Community of Aldrichina grahami (Diptera: Calliphoridae) across Developmental Stages.

The blowfly, Aldrichina grahami (Diptera: Calliphoridae), is a well-known forensically important insect. Basic data related to A. grahami have been well documented; but despite the pivotal role of gut microbes in various facets of insect biology, little is known about its gut microbiome. To investigate the gut bacterial community of A. grahami and explore its stability, diet and developmental stage were selected as the two variables. Larvae were reared on bovine liver, swine manure, and chicken manure, and high-throughput sequencing of the 1st, 2nd, and 3rd instar larvae and the newly emerged adults was performed. According to our results, the alpha diversity of the gut bacterial community did not significantly vary among different diets and developmental stages. Principal coordinate analysis revealed that the gut microbiome of A. grahami clustered together among different diets and developmental stages. The main phyla in the gut microbial community of A. grahami were Firmicutes and Proteobacteria, and the dominant genera were Vagococcus, Providencia, Lactobacillus, and Morganella. These findings characterized the gut microbiome of A. grahami and demonstrated that the gut bacterial community is fairly stable. The dominant genera Vagococcus, Providencia, Lactobacillus, and Morganella have the potential to serve as core microbiomes within the gut microbial community of A. grahami.

Polystyrene microplastics protect lettuce (Lactuca sativa) from the hazardous effects of Cu(OH)2 nanopesticides.

Copper-based nanopesticides are released into the environment during foliar spray application, and they could, on their own or in combination with microplastics (MPs), pose threats to environmental safety and human health. In this study, Cu(OH)2 nanowires greatly decreased the vigor of lettuce seeds (p< 0.01) and the root length of lettuce seedlings (p< 0.01) and significantly altered the lettuce antioxidant defence system and MDA content (p< 0.05). Released Cu2+ played a critical role in the toxicity mechanism of Cu(OH)2 nanowires in lettuce seedlings, as evidenced by the substantial accumulation of Cu in the seedling roots (p< 0.01) rather than in the leaves. Polystyrene (PS) MPs (1 mg/L) stimulated lettuce seedling growth, as shown by the (highly) significant increase in root and leaf length and in the seed vigor index (p< 0.01 or 0.05). Notably, PS MPs (1 mg/L) neutralized the hazardous effects of 1 mg/L Cu(OH)2 nanowire treatment on lettuce growth, as reflected by the vitality and root length of the seedlings returning to normal levels. The PS MPs (1 mg/L) absorbed on middle root surfaces and strongly hindered Cu accumulation in lettuce roots, which was the predominant mechanism by which PS MPs suppressed the hazardous effects of the Cu(OH)2 nanowires. This study strengthens the understanding of the toxicity and toxicity mechanisms of Cu(OH)2 nanowires with or without PS MPs in the environment.

The Effects of Diets and Long-term Laboratory Rearing on Reproduction, Behavior, and Morphology of Lucilia cuprina (Diptera: Calliphoridae).

The Australian sheep blow fly, Lucilia cuprina (Wiedemann), is commonly reared in the laboratory for many sequential generations on simple, fixed diets, so it can be used in veterinary, medical, and forensic studies. To investigate the effect of diet and long-term laboratory rearing on L. cuprina, flies were fed with two different diets (sugar and milk-sugar) over a year and F1, F6, and F11 generations were used for comparisons based on the number of eggs, attraction to wool and liver, and wing size. The results showed that the number of eggs of gravid flies, and the attractiveness of wool and liver did not differ significantly between diets and generations, but gravid flies were more attracted to wool and liver than non-gravid flies (P < 0.05). Moreover, in the F1 generation, thorax length and wing aspect ratio were significantly longer than in the F6 and F11 generations (P < 0.05), and the wing length was significantly longer than in the F11 generation (P < 0.05). It was concluded that neither diet nor long-term laboratory rearing affect potential fecundity or the behavioral responses of L. cuprina, but the gravidity of flies affects their behavioral response, and long-term laboratory rearing significantly affects fly morphology, apparently explaining a loss in flight performance.

Behavior and Electrophysiological Response of Gravid and Non-Gravid Lucilia cuprina (Diptera: Calliphoridae) to Carrion-Associated Compounds.

The Australian blow fly, Lucilia cuprina Wiedmann (Diptera: Calliphoridae), is a major cause of myiasis (flystrike) in Merino sheep in Australia and New Zealand and, as a primary colonizer of fresh carrion, also an important species in forensic investigations. Olfaction is considered the most important cue for insects to rapidly locate carrion over long distances, so the first carrion visitors are predicted to be very sensitive to carrion-related volatile compounds. We studied the responses of the Australian blow fly, Lucilia cuprina, to the carrion-associated compounds dimethyl trisulfide (DMTS), butyric acid, 1-octen-3-ol and indole. We also tested 2-mercaptoethanol, a compound commonly used in fly traps in Australia. We investigated whether responses of the flies are affected by their ovarian status by comparing responses of gravid and non-gravid L. cuprina in electroantennography (EAG) and two-choice laboratory bioassays. All four compounds evoked an EAG response, while only DMTS evoked responses in gas chromatography-mass spectrometry electroantennographic detection (GCMS-EAD) analyses and two-choice bioassays. Gravid flies detected lower doses of the test compounds than non-gravid flies. Our results indicate that DMTS is an important semiochemical for L. cuprina to locate carrion resources, and has potential for use in fly traps for flystrike control. Our observations also suggest that the greater sensitivity of gravid L. cuprina allows them to find fresh carrion quickly to maximize reproductive success by avoiding unsuitable degraded carrion.

Association of heat shock protein 90 with motility of post-thawed sperm in bulls.

The correlation between the 90 kDa heat-shock protein (HSP90) and sperm quality following the process of freezing-thawing in bulls has not been studied clearly. Therefore, the objective of the present was to clarify the relationship between HSP90 level and semen parameters during the process of cryopreservation in bulls. Semen samples from 5 Holstein bulls were obtained by artificial vagina. Characteristics of these semen at three stages (fresh, after equilibration and frozen-thawed), including motility, plasma membrane integrity and acrosome integrity were evaluated. The mRNA expression level of HSP90 at the three stages was evaluated by using quantitative Real-Time PCR. Meanwhile, the protein level of HSP90 expression at the three stages was detected according to Western blot. The results showed that sperm parameters evaluated in fresh semen was the highest in the three groups. Sperm parameters in semen after equilibration were lower than those in fresh semen (P>0.05) and higher than those in post-thawed semen (P<0.05). Sperm parameters in frozen-thawed semen were the lowest among the three groups (P<0.05). This study indicated that HSP90 expression is proportional to sperm quality. HSP90 expression level in fresh semen was significantly higher than that in frozen-thawed semen (P<0.05). Although no significant differences in HSP90 expression were observed between fresh semen and semen after equilibration (P>0.05). Results in this study suggest that HSP90 level in bull spermatozoa was gradually declined following the process of freezing-thawing, and might be associated with sperm motility, plasma membrane integrity and acrosome integrity.

Interspecific variation in hypoxia tolerance, swimming performance and plasticity in cyprinids that prefer different habitats.

This study quantified and compared hypoxia tolerance and swim performance among cyprinid fish species from rapid-, slow- and intermediate-flow habitats (four species per habitat) in China. In addition, we explored the effects of short-term acclimation on swim performance, maximum metabolic rate (M(O2,max)) and gill remodelling to detect habitat-associated patterns of plastic response to hypoxia. Indices of hypoxia tolerance included oxygen threshold for loss of equilibrium (LOE50) and aquatic surface respiration (ASR50), and critical oxygen tension for routine metabolic rate (Pcrit). Critical swimming speed (Ucrit) and M(O2,max) were measured under normoxic and hypoxic conditions after 48 h acclimation to normoxia and hypoxia, and gill remodelling was estimated after 48 h of hypoxia exposure. Both traditional ANCOVA and phylogenetically independent contrast (PDANOVA) analyses showed that fish species from rapid-flow habitats exhibited lower LOE50 compared with fish from intermediate- and slow-flow habitats. Habitat-specific differences in Pcrit and Ucrit were detected using PDANOVA but not traditional ANCOVA analyses, with fish species from rapid-flow habitats exhibiting lower Pcrit but higher Ucrit values compared with fish from intermediate- and slow-flow habitats. Fish species from rapid-flow habitats were also characterized by less plasticity in swim performance and gill morphology in response to hypoxia acclimation compared with species from slow-flow habitats, but a greater drop in swim performance in response to acute hypoxia exposure. The study detected a habitat-specific difference in hypoxia tolerance, swimming performance and its plasticity among fish from habitats with different flow conditions, possibly because of the long-term adaptation to the habitat caused by selection stress. The PDANOVA analyses were more powerful than traditional statistical analyses according to the habitat effects in both hypoxia tolerance and swimming performance in this study.

Integrating environmental variation, predation pressure, phenotypic plasticity and locomotor performance.

The Wujiang River, a tributary of the Three Gorges Reservoir, has many dams along its length. These dams alter the river's natural habitat and produce various flow regimes and degrees of predator stress. To test whether the swimming performance and external body shape of pale chub (Zacco platypus) have changed as a result of alterations in the flow regime and predator conditions, we measured the steady (U(crit)) and unsteady (fast-start) swimming performances and morphological characteristics of fish collected from different sites along the Wujiang River. We also calculated the maximum respiratory capacity and cost of transport (COT). We demonstrated significant differences in swimming performance and morphological traits among the sampling sites. Steady swimming performance was positively correlated with water velocity and negatively correlated with the abundance of predators, whereas unsteady swimming performance was negatively correlated with water velocity. The body shape was significantly correlated with both swimming performance and ecological parameters. These findings suggested that selection pressure on swimming performance results in a higher U(crit) and a more streamlined body shape in fast-flow and (or) in habitats with low predator stress and subsequently results in a lower COT. These characteristics were accompanied by a poorer fast-start performance than that of the fish from the slow-flow and (or) high-predator habitats. The divergence in U(crit) may also be due in part to variation in respiratory capacity.

The effects of fasting on the metabolic interaction between digestion and locomotion in juvenile southern catfish (Silurus meridionalis).

To investigate the effect of fasting on maintenance metabolism, feeding metabolism and aerobic swimming performance as well as their metabolic interactions in juvenile southern catfish, we measured the following: (1) the postprandial oxygen consumption ((MO2) response (16% body mass meal size) after 0 (control), 1, 2 and 4 weeks of fasting and (2) the swimming performance of non-digesting and digesting fish after either 0, 1, 2 or 4 weeks of fasting. The fasting groups displayed with lower resting MO2 (MO2(rest)), lower peak postprandial MO2 (MO2(peak)), larger energy expenditures and longer digestive processes than those of the control groups. The critical swimming speed (U(crit)), the active MO2 (MO2(active)) and the metabolic scope (MO2(active) - MO2(rest), MS) of both non-digesting and digesting fish all decreased progressively after 1, 2 and 4 weeks of fasting, with those of non-digesting fish decreased more acutely than digesting fish (P<0.05). Digesting fish displayed with a 14%, 23%, 27% and 71% significantly higher MO2(active) than that of non-digesting fish in the 0-, 1-, 2- and 4-week fasting groups. Digestion only caused a significantly lower U(crit) and MS in both the 0- and 1-week fasting groups (P<0.05). The MO2 increased greatly with the swimming speed, and digestion caused a higher MO2 when compared to that of the fasting fish in any groups. The MO2 of fish in the 4-week fasting group was significantly lower than that of other groups when the swimming speed was the same. In conclusion, both digestive and locomotive functions were down-regulated during fasting. In the 0- and 1-week fasting groups, the decreased MS for swimming during digestion caused a lower U(crit) (i.e., a digestion priority model). However, because the MO2(active) of digesting fish decreased much more slowly than that of fasting fish, the MS of the 2- and 4-week fasting groups did not change during digestion, and the fish could handle both physiological activities independently (i.e., an additive model).