Laser-based killing of a macroparasite inside its live invertebrate host

Whether host phenotypic alterations induced by parasites are reversible or not is a core issue to understand underlying mechanisms as well as the fitness costs of infection and recovery to the host. Clearing infection is an essential step to address this issue, which turns out to be challenging with endoparasites of large size relative to that of their host. Here, we took advantage of the lethality, contactless and versatility of high-energy laser beam to design such tool, using thorny-headed worms and their amphipod intermediate host as a model system. We show that laser-based de-parasitization can be achieved using blue laser targeting carotenoid pigments in Polymorphus minutus but not in the larger and less pigmented Pomphorhynchus tereticollis. Using DNA degradation to establish parasite death, we found that 80% P. minutus died from within-host laser exposure. Survival rate of infected gammarids to laser treatment was higher than uninfected ones. The failure to kill P. tereticollis was also observed with nanosecond-green laser, an alternative laser source targeting lipid. We discuss the possible causes of amphipod death following parasite treatment and highlight the perspectives that this technology offers

Laser-based killing of a macroparasite inside its live invertebrate host

Clearing infection is an essential step to address many issues in host-parasite interactions but is challenging when dealing with endoparasites of large size relative to that of their host. Here, we took advantage of the lethality, contactless and versatility of high-energy laser beam to achieve it, using thorny-headed worms (Acanthocephala) and their amphipod intermediate host as a model system. We show that laser-based de-parasitization can be achieved using 450 nm Blue Diode Laser targeting carotenoid pigments in the bird acanthocephalan Polymorphus minutus. Using proboscis evagination failure and DNA degradation to establish parasite death, we found that 80% P. minutus died from within-host exposure to 5 pulses of 50 ms duration, 1.4 W power. Survival of infected gammarids 11 days after laser treatment was 60%. Preliminary tests were also performed with Nanosecond-Green Laser targeting lipids in Pomphorhynchus tereticollis, another acanthocephalan parasite. We discuss the efficiency and side-effect of laser treatment in this host-parasite system and highlight the perspectives that this technology more generally offers in parasitology

“Born with a silver spoon in the mouth has bad sides too”: Experimentally increasing growth rate enhances individual quality but accelerates reproductive senescence in females of the mealworm beetle, Tenebrio molitor

International audienceSenescence occurs because of the decline of the strength of selection with age, allowing late-life reduced performances not being counter selected. From there, several phenomena may explain late-life reduced performances, such as the accumulation of deleterious mutations, the expression of pleiotropic genes or the existence of resource trade-offs between early and late performances. This latter phenomenon is at the core of the disposable soma theory of aging, which predicts that growth and early-life reproduction have costs that increase reproductive and actuarial senescence. Whereas the impact of the cost of early reproduction on reproductive and actuarial senescence has been extensively studied, that of the cost of growth remains overlooked and often inconclusive, possibly because of confounding effects associated with the procedures used to manipulate growth rate. Here, we investigated the cost of growth rate and its impact on reproductive senescence and longevity of females of the mealworm beetle, Tenebrio molitor. For this purpose, we generated insects with contrasted growth rates by raising groups of them in conditions below, above and optimal relative humidity (RH: 55, 85 and 70%, respectively) during the larval stage. Theresulting adult females then bred, under the same optimal RH conditions, early in life, then later in life and were followed there until death. We found that larvae grown under the highest relative humidity exhibited the highest larval growth rate, thanks to both shorter growth duration and the achievement of heavier pupae mass. Adult females from this favorable growing condition lived longer, were more fecund early in life, but suffered from lower late-life reproductive investment. Our study shows that growth rate, which is highly dependent on the early-life environment, is an important factor modulating adult reproductive senescence, through the occurrence of early-late life trade-offs

Optimization of anesthetic procedure in crustaceans: evidence for sedative and analgesic-like effect of MS-222 using a semi-automated device for exposure to noxious stimulus.

10 pagesInternational audienceThe implementation of anesthetic procedure in aquatic crustaceans remains mostly limited to studies dealing with sedation and survival from anesthesia, possibly owing to the debated question of pain in invertebrates. However, two important issues are generally overlooked: actual analgesic-like effect, and possible physiological post-anesthesial effects. Here we report on the anesthetic properties and possible after-effects of MS-222 (Tricaine Methanesulfonate or Ethyl 3-aminobenzoate methanesulfonate) and Eugenol in the freshwater amphipod Gammarus pulex. We first optimized the concentration of MS-222, and the induction and recovery time, based on preliminary tests and published studies. We then relied on the nociceptive modulation of sheltering behavior to assess the analgesic-like effect of the two drugs, using a new semi-automated electric shock device. In addition, we monitored the impact of anesthesia with MS-222 on locomotor activity and oxygen consumption and addressed potential adverse effects upon recovery using biomarkers related to metabolism and neurotoxicity. We provide evidence for the sedative and analgesic-like effects of MS-222 at 600 mg.L-1 and, to a lesser extent, of Eugenol at 100 µL.L-1, with no decrease in survival rate at 6 days post anesthesia. Oxygen consumption was reduced -but not eliminated- under full anesthesia with 600 mg.L-1 MS-222. No significant physiological effect of anesthesia was evidenced on the activity of the mitochondrial electron transfer system, or that of acetylcholine esterase, nor on total antioxidant capacity. We therefore conclude to the efficiency of MS-222 as an anesthetic drug in G. pulex. Eugenol should be tested at a higher concentration to reach the same efficiency, providing that increased concentration would not incur side-effects. Furthermore, the new and original semi-automated electric chock device used to induce nociception can be easily adapted to any species of aquatic invertebrates and small-sized fish and tadpoles, offering a standardized and flexible protocol to study nociceptive response and anesthesia in aquatic organisms

A dietary carotenoid reduces immunopathology and enhances longevity through an immune depressive effect in an insect model

Abstract Immunopathology corresponds to self-damage of the inflammatory response, resulting from oxidizing molecules produced when the immune system is activated. Immunopathology often contributes to age-related diseases and is believed to accelerate ageing. Prevention of immunopathology relies on endogenous antioxidant enzymes and the consumption of dietary antioxidants, including carotenoids such as astaxanthin. Astaxanthin currently raises considerable interest as a powerful antioxidant and for its potential in alleviating age-related diseases. Current in vitro and short-term in vivo studies provide promising results about immune-stimulating and antioxidant properties of astaxanthin. However, to what extent dietary supplementation with astaxanthin can prevent long-term adverse effects of immunopathology on longevity is unknown so far. Here, using the mealworm beetle, Tenebrio molitor, as biological model we tested the effect of lifetime dietary supplementation with astaxanthin on longevity when exposed to early life inflammation. While supplementation with astaxanthin was found to lessen immunopathology cost on larval survival and insect longevity, it was also found to reduce immunity, growth rate and the survival of non immune-challenged larvae. This study therefore reveals that astaxanthin prevents immunopathology through an immune depressive effect and can have adverse consequences on growth

Host manipulation revisited: no evidence for a causal link between altered photophobia and increased trophic transmission of amphipods infected with acanthocephalans.

8 pagesInternational audience1. Many parasites with complex life cycles critically rely on trophic transmission to pass from an intermediate host to a final host. Parasite-induced behavioural alterations in intermediate hosts are often supposed to be adaptive through increasing the susceptibility of intermediate hosts to predation by final hosts. However, the evidence is so far only correlational, and direct evidence for a causal link between one single behavioural alteration and increased trophic transmission is still missing. 2. Here, we addressed, for the first time, the relationship between increased vulnerability to fish predation and altered photophobia in an amphipod, Gammarus pulex, infected with a fish acanthocephalan, Pomphorhynchus tereticollis. 3. In microcosms, naturally-infected amphipods were significantly more vulnerable to fish predation than uninfected ones at two different light intensities. However, although variation in illumination significantly affected the extent of difference in photophobia between infected and uninfected individuals, it had no effect on predation bias towards infected amphipods. 4. In addition, although an injection of a mixture of serotonin and fluoxetine in uninfected amphipods mimicked the parasite-induced decreased photophobia, injected individuals were not more vulnerable to fish predation that uninfected ones injected with a control solution. 5. Overall, our results indicate that the decreased photophobia in infected intermediate hosts does not play in itself a causal role in the trophic transmission of the parasite to its final host. The actual role a parasite-induced behavioural alteration plays in trophic transmission should be carefully assessed before an adaptive interpretation is given

DataSheet_1_Infection risk by oral contamination does not induce immune priming in the mealworm beetle (Tenebrio molitor) but triggers behavioral and physiological responses.docx

In invertebrates, immune priming is the ability of individuals to enhance their immune response based on prior immunological experiences. This adaptive-like immunity likely evolved due to the risk of repeated infections by parasites in the host’s natural habitat. The expression of immune priming varies across host and pathogen species, as well as infection routes (oral or wounds), reflecting finely tuned evolutionary adjustments. Evidence from the mealworm beetle (Tenebrio molitor) suggests that Gram-positive bacterial pathogens play a significant role in immune priming after systemic infection. Despite the likelihood of oral infections by natural bacterial pathogens in T. molitor, it remains debated whether ingestion of contaminated food leads to systemic infection, and whether oral immune priming is possible is currently unknown. We first attempted to induce immune priming in both T. molitor larvae and adults by exposing them to food contaminated with living or dead Gram-positive and Gram-negative bacterial pathogens. We found that oral ingestion of living bacteria did not kill them, but septic wounds caused rapid mortality. Intriguingly, the consumption of either dead or living bacteria did not protect against reinfection, contrasting with injury-induced priming. We further examined the effects of infecting food with various living bacterial pathogens on variables such as food consumption, mass gain, and feces production in larvae. We found that larvae exposed to Gram-positive bacteria in their food ingested less food, gained less mass and/or produced more feces than larvae exposed to contaminated food with Gram-negative bacteria or control food. This suggests that oral contamination with Gram-positive bacteria induced both behavioral responses and peristalsis defense mechanisms, even though no immune priming was observed here. Considering that the oral route of infection neither caused the death of the insects nor induced priming, we propose that immune priming in T. molitor may have primarily evolved as a response to the infection risk associated with wounds rather than oral ingestion.</p