Challenging Popular Belief, Mosquito Larvae Breathe Underwater.

Immature mosquitoes are thought to breathe only atmospheric air through their siphons despite reports of prolonged submerged survival. We studied the survival of last-instar larvae of Aedes aegypti fully submerged at different temperatures and measured the oxygen consumption from air and water-dissolved larvae and pupae of this species under different conditions. Larvae survived much longer than expected, reaching 50% mortality only after 58, 10, and 5 days at 15°, 25°, and 35 °C, respectively. Larval to pupa molt was only observed in larvae with access to air, whereas individuals kept submerged never molted. Although most of the oxygen was obtained from the air, larvae obtained 12.72% of their oxygen from the water, while pupae took only 5.32%. In both media, temperature affected the respiration rate of the larvae, with relatively close Q10 values (1.56 and 1.83 for water and air, respectively). A similar pattern of O2 consumption was observed in Ae. albopictus, whose larvae obtained 12.14% of their oxygen from the water. The detailed quantification of oxygen consumption by mosquito larvae showed that water-dissolved oxygen is not negligible and physiologically relevant, challenging the idea that mosquito larvae only breathe atmospheric oxygen.

Effect of host age, sex and life stage on the prevalence and abundance of sucking lice on Weddell seal in the Antarctic Peninsula.

Through evolutionary time, seal lice have developed morphological, behavioral, and ecological adaptations to cope with the amphibious lifestyle of their hosts in a co-evolutionary process. Consequently, the dynamics of lice populations are determined by seals behavior. We aim to study the effects of host sex, age class, year, and sampling location, on the prevalence and mean abundance of Antarctophthirus carlinii, on Weddell seals (WS) Leptonychotes wedelli. The study was conducted at two sites in the Antarctic Peninsula, namely, Marambio/Seymour Island (MI) and the Danco Coast (DC). We collected lice from 71 WS: 33 from MI, during the reproductive season, and 38 from DC, during the molting season, between 2014 and 2017. According to our analyses, host age class and sex were the variables that affected prevalence levels of lice on WS. Whereas, age class, year, site, and sex affected lice mean abundance. Juveniles presented higher prevalence and mean abundance than adults, possibly acting as reservoirs for lice as they move through different colonies until they reach reproductive age. Concurrently, seals during molting season were more infested. Unlike nursing, during the molting season seals spend much time ashore forming mixed groups that favor both egg development and lice transmission.

How Did Seal Lice Turn into the Only Truly Marine Insects?

Insects are the most evolutionarily and ecologically successful group of living animals, being present in almost all possible mainland habitats; however, they are virtually absent in the ocean, which constitutes more than 99% of the Earth's biosphere. Only a few insect species can be found in the sea but they remain at the surface, in salt marshes, estuaries, or shallow waters. Remarkably, a group of 13 species manages to endure long immersion periods in the open sea, as well as deep dives, i.e., seal lice. Sucking lice (Phthiraptera: Anoplura) are ectoparasites of mammals, living while attached to the hosts' skin, into their fur, or among their hairs. Among them, the family Echinophthiriidae is peculiar because it infests amphibious hosts, such as pinnipeds and otters, who make deep dives and spend from weeks to months in the open sea. During the evolutionary transition of pinnipeds from land to the ocean, echinophthiriid lice had to manage the gradual change to an amphibian lifestyle along with their hosts, some of which may spend more than 80% of the time submerged and performing extreme dives, some beyond 2000 m under the surface. These obligate and permanent ectoparasites have adapted to cope with hypoxia, high salinity, low temperature, and, in particular, conditions of huge hydrostatic pressures. We will discuss some of these adaptations allowing seal lice to cope with their hosts' amphibious habits and how they can help us understand why insects are so rare in the ocean.

Under pressure: the extraordinary survival of seal lice in the deep sea.

Lice from pinnipeds - sea lions, seals and walruses - are the only insects capable of surviving marine dives. Throughout their evolutionary history, they have adapted to tolerate hypoxia, high salinity, low temperature and, in particular, to tolerate conditions of high hydrostatic pressure. To understand the limits of the capacity of lice to survive during host deep dives, we conducted a series of controlled experiments in the laboratory. We collected lice from elephant seals and submitted the different life stages to high pressure conditions. Lice were first exposed to one of four hydrostatic pressures: 30, 80, 150 or 200 kg cm-2 They were then exposed a second time to higher or lower hydrostatic pressure conditions to test for the impact of the first experience, which could either be deleterious or trigger physiological adaption, allowing them a better tolerance to high pressure. We found that lice from elephant seals can tolerate hydrostatic pressures higher than 200 kg cm-2 (close to 200 atm), which is equivalent to 2000 m depth. Adults exhibited lower recovery times than nymphs after immersion at high hydrostatic pressure. Our findings show that lice have developed unique adaptations to endure extreme marine conditions. We discuss these extreme performances in relation to the morphological characteristics and physiological responses to diving in these insects.

Corrigendum to "Lousy chicks: Chewing lice from the imperial shag, Leucocarbo atriceps" [Int. J. Parasitol. Parasites Wildl. 6 (2017) 229-232].

[This corrects the article DOI: 10.1016/j.ijppaw.2017.08.002.].

Lousy chicks: Chewing lice from the Imperial Shag, Leucocarbo atriceps.

Forty-one imperial shag chicks were sampled for lice during the breeding season of 2014 in Punta León, Argentina. We found 2 lice species, Pectinopygus turbinatus infesting the body and Piagetiella caputincisum present in the oral cavity of the birds. This constitutes the first host record for P. turbinatus and the first record for the continental Argentina for P. caputincisum. Ninety-three percent of the chicks were infested by at least one lice species. P. turbinatus was present in all of the lousy chicks, while P. caputincisum infested 84.2% of them. The mean intensity was 29.5 and the range 1-129. There was no difference in prevalence, mean intensity or mean abundance between louse species. However, we found differences among the pattern of infestation of each species. Imperial shag chicks were infested by their parents during their first days of life by P. turbinatus, mainly in nymphal stage and by P. caputincisum as adult lice. Our results showed differences among lice species that could be related to the restrictions that lice from seabirds faced during their life cycle.

What does heat tell a mosquito? Characterization of the orientation behaviour of Aedes aegypti towards heat sources.

The use of heat as a cue for the orientation of haematophagous insects towards hot-blooded hosts has been acknowledged for many decades. In mosquitoes, thermoreception has been studied at the molecular, physiological and behavioural levels, and the response to heat has been evaluated in multimodal contexts. However, a direct characterization of how these insects evaluate thermal sources is still lacking. In this study we characterize Aedes aegypti thermal orientation using a simple dual choice paradigm, providing direct evidence on how different attributes of heat sources affect their choice. We found that female mosquitoes, but not males, are able to discriminate among heat sources that are at ambient, host-range and deleterious temperatures when no other stimuli are present, eliciting a positive response towards host-range and an avoidance response towards deleterious temperatures. We also tested the preference of females according to the size and position of the sources. We found that females do not discriminate between heat sources of different sizes, but actively orientate towards closer sources at host temperature. Furthermore, we show that females cannot use IR radiation as an orientation cue. Orientation towards a host involves the integration of cues of different nature in distinct phases of the orientation. Although such integration might be decisive for successful encounter of the host, we show that heat alone is sufficient to elicit orientation behaviour. We discuss the performance of mosquitoes' thermal behaviour compared to other blood-sucking insects.

Uncovering deep mysteries: the underwater life of an amphibious louse.

Despite the incredible success of insects in colonizing almost every habitat, they remain virtually absent in one major environment--the open sea. A variety of hypotheses have been raised to explain why just a few insect species are present in the ocean, but none of them appears to be fully explanatory. Lice belonging to the family Echinophthiriidae are ectoparasites on different species of pinnipeds and river otters, i.e. they have amphibious hosts, who regularly perform long excursions into the open sea reaching depths of hundreds of meters (thousands of feets). Consequently, lice must be able to support not only changes in their surrounding media, but also extreme variations in hydrostatic pressure as well as breathing in a low oxygen atmosphere. In order to shed some light on the way lice can survive during the diving excursions of their hosts, we have performed a series of experiments to test the survival capability of different instars of Antarctophthirus microchir (Phthiraptera: Anoplura) from South American sea lions Otaria flavescens, when submerged into seawater. These experiments were aimed at analyzing: (a) immersion tolerance along the louse life; (b) lice's ability to obtain oxygen from seawater; (c) physiological responses and mechanisms involved in survival underwater. Our experiments showed that the forms present in non-diving pups--i.e. eggs and first-instar nymphs--were unable to tolerate immersion in water, while following instars and adults, all usually found in diving hosts, supported it very well. Furthermore, as long as the level of oxygen dissolved in water was higher, the lice survival capability underwater increased, and the recovery period after returning to air declined. These results are discussed in relation to host ecology, host exploitation and lice functional morphology.

Antarctophthirus carlinii (Anoplura: Echinophthiriidae), a new species from the Weddell seal Leptonychotes weddelli.

As a part of an ongoing long-term study on the biology of pack-ice seals in Antarctica, we had the opportunity to collect lice from Weddell seals (Leptonychotes weddelli). We did not find the original description of this host-parasite association. Antarctophthirus ogmorhini had previously been reported as a parasite for the Weddell seal, but the information is, to a certain extent, confusing. During the development of the present study, we had access to literature concerning the presence of A. ogmorhini on this host, which, to our knowledge, was not determined in any of the previous works on this species. We compared lice collected from Weddell seals with A. ogmorhini obtained from the type host, the leopard seal (Hydrurga leptonyx), and we found that both species can be distinguished. The main differences are the characteristic pattern of chaetotaxy in the dorsal side of the head in lice from Weddell seals, the size and form of the pseudopenis, and the distribution and size of the fringe of setae surrounding the genital opening. Considering the conservative morphology, and ecological and evolutionary features of sucking lice, we proposed that lice from Weddell seals constitute a new species. In the present work, we described and illustrated adults of this new species collected from Weddell seals during the austral summer of 2014 at the Danco Coast, Antarctic Peninsula.

Faster the better: a reliable technique to sample anopluran lice in large hosts.

Among Anoplura, the family Echinophthiriidae includes those species that infest mainly the pinnipeds. Working with large hosts implies methodological considerations as the time spent in the sampling, and the way in that the animal is restrained. Previous works on echinophthiriids combined a diverse array of analyses including field counts of lice and in vitro observations. To collect lice, the authors used forceps, and each louse was collected individually. This implied a long manipulation time, i.e., ≈60 min and the need to physically and/or chemically immobilize the animal. The present work described and discussed for the first a sample technique that minimized the manipulation time and also avoiding the use of anesthesia. This methodology implied combing the host's pelage with a fine-tooth plastic comb, as used in the treatment of human pediculosis, and keeping the comb with the lice retained in a Ziploc® bag with ethanol. This technique was used successfully in studies of population dynamic, habitat selection, and transmission pattern, being a reliable methodology. Lice are collected entirely and are in a good condition to prepare them for mounting for studying under light or scanning electron microscopy. Moreover, the use of the plastic comb protects from damaging taxonomically important structures as spines being also recommended to reach taxonomic or morphological goals.

Scanning electron microscopy of Antarctophthirus microchir (Phthiraptera: Anoplura: Echinophthiriidae): studying morphological adaptations to aquatic life.

The members of the Family Echinophthiriidae (Phthiraptera: Anoplura) are unique among insects because they infest hosts with an amphibious lifestyle. During their evolution they developed morphological traits that are reflected in unique features. The SEM is a helpful tool to analyze them. Knowing in detail the external structure of these lice is the first step to understand the whole process that derived from the co-adaptation of lice and pinnipeds to the marine environment. For the first time, we studied the external structure of all stages of an echinophthiriid louse. The results are discussed in the light of their evolutionary, functional, and ecological implications.