A large West Antarctic Ice Sheet explains early Neogene sea-level amplitude.

Early to Middle Miocene sea-level oscillations of approximately 40-60 m estimated from far-field records1-3 are interpreted to reflect the loss of virtually all East Antarctic ice during peak warmth2. This contrasts with ice-sheet model experiments suggesting most terrestrial ice in East Antarctica was retained even during the warmest intervals of the Middle Miocene4,5. Data and model outputs can be reconciled if a large West Antarctic Ice Sheet (WAIS) existed and expanded across most of the outer continental shelf during the Early Miocene, accounting for maximum ice-sheet volumes. Here we provide the earliest geological evidence proving large WAIS expansions occurred during the Early Miocene (~17.72-17.40 Ma). Geochemical and petrographic data show glacimarine sediments recovered at International Ocean Discovery Program (IODP) Site U1521 in the central Ross Sea derive from West Antarctica, requiring the presence of a WAIS covering most of the Ross Sea continental shelf. Seismic, lithological and palynological data reveal the intermittent proximity of grounded ice to Site U1521. The erosion rate calculated from this sediment package greatly exceeds the long-term mean, implying rapid erosion of West Antarctica. This interval therefore captures a key step in the genesis of a marine-based WAIS and a tipping point in Antarctic ice-sheet evolution.

Ancient Human Genomes and Environmental DNA from the Cement Attaching 2,000-Year-Old Head Lice Nits.

Over the past few decades, there has been a growing demand for genome analysis of ancient human remains. Destructive sampling is increasingly difficult to obtain for ethical reasons, and standard methods of breaking the skull to access the petrous bone or sampling remaining teeth are often forbidden for curatorial reasons. However, most ancient humans carried head lice and their eggs abound in historical hair specimens. Here we show that host DNA is protected by the cement that glues head lice nits to the hair of ancient Argentinian mummies, 1,500-2,000 years old. The genetic affinities deciphered from genome-wide analyses of this DNA inform that this population migrated from north-west Amazonia to the Andes of central-west Argentina; a result confirmed using the mitochondria of the host lice. The cement preserves ancient environmental DNA of the skin, including the earliest recorded case of Merkel cell polyomavirus. We found that the percentage of human DNA obtained from nit cement equals human DNA obtained from the tooth, yield 2-fold compared with a petrous bone, and 4-fold to a bloodmeal of adult lice a millennium younger. In metric studies of sheaths, the length of the cement negatively correlates with the age of the specimens, whereas hair linear distance between nit and scalp informs about the environmental conditions at the time before death. Ectoparasitic lice sheaths can offer an alternative, nondestructive source of high-quality ancient DNA from a variety of host taxa where bones and teeth are not available and reveal complementary details of their history.

Human follicular mites: Ectoparasites becoming symbionts.

Most humans carry mites in the hair follicles of their skin for their entire lives. Follicular mites are the only metazoans tha continuously live on humans. We propose that Demodex folliculorum (Acari) represents a transitional stage from a host-injuring obligate parasite to an obligate symbiont. Here, we describe the profound impact of this transition on the genome and physiology of the mite. Genome sequencing revealed that the permanent host association of D. folliculorum led to an extensive genome reduction through relaxed selection and genetic drift, resulting in the smallest number of protein-coding genes yet identified among panarthropods. Confocal microscopy revealed that this gene loss coincided with an extreme reduction in the number of cells. Single uninucleate muscle cells are sufficient to operate each of the three segments that form each walking leg. While it has been assumed that the reduction of the cell number in parasites starts early in development, we identified a greater total number of cells in the last developmental stage (nymph) than in the terminal adult stage, suggesting that reduction starts at the adult or ultimate stage of development. This is the first evolutionary step in an arthropod species adopting a reductive, parasitic or endosymbiotic lifestyle. Somatic nuclei show underreplication at the diploid stage. Novel eye structures or photoreceptors as well as a unique human host melatonin-guided day/night rhythm are proposed for the first time. The loss of DNA repair genes coupled with extreme endogamy might have set this mite species on an evolutionary dead-end trajectory.

Mesostigmata diversity by manure type: a reference study and new datasets from southwestern Iran.

Manure-inhabiting Mesostigmata mites are important biological control agents of pest flies. However, the biodiversity of this mite community is mainly known from Europe and America, and especially from cattle manure. This study examined the diversity and abundance of Mesostigmata mites associated with various types of manure in an (intensive) agricultural region of the Middle East, i.e., the city Ahvaz and its suburbs, in southwest Iran. Mite samples were extracted from manure of cattle, buffalo, sheep, horse, poultry and quail in 30 livestock and poultry farms. In total, 40 species belonging to 24 genera and 16 families were identified. The most diverse families were Laelapidae with eight species, Macrochelidae with seven and Parasitidae with six. Macrocheles muscaedomesticae and Uroobovella marginata were the most widespread species, recorded in 28 and 27 out of 30 collection sites, respectively. Two species, M. sumbaensis and U. marginata, were found in all studied manures. Simpson's diversity index recorded the highest diversity in buffalo and sheep manure. Real and theoretical species richness (rarefaction curves) were congruent in number of individuals. The presence of seven species of Macrochelidae in the manure confirms that these are important predators of the house fly for the region of Ahvaz and its suburbs. Members of the Parasitidae were highly prevalent, with one species known as a specialized predator of house fly eggs. This work aims to encourage further studies on the diversity of Mesostigmata in these agricultural settings, and further continue assessing the feasibility of these mites as effective biocontrol agents of filth flies in different types of manure and from different corners of the world.

Assemblages of Acari in shallow burials: mites as markers of the burial environment, of the stage of decay and of body-cadaver regions.

The burial of a cadaver results in reduced arthropod activity and disruptions in colonisation patterns. Here, the distribution and diversity of mite taxa was studied across decomposition stages of shallowly buried pig carcasses (Sus scrofa domesticus). In total 300 mites (88 species) were collected from three pig shallow graves compared to 129 mites (46 species) from control (bare) soil samples at the same depth. A successional pattern of Acari higher taxa and families was observed, and species richness and biodiversity fluctuated throughout decomposition, whereas active decay showed the greatest biodiversity. Mesostigmata mites were the most abundant in 'cadaver soils' with a significant difference in the abundance of Parasitidae mites, whereas Oribatida mites (true soil mites) were the most abundant in control soils. Certain mite species were significantly associated with decay stages: Cornigamasus lunaris with 'bloated', Gamasodes spiniger with 'active', Eugamasus sp. and Lorryia reticulata with 'advanced', and Macrocheles matrius and Ramusella clavipectinata in 'dry'. Scheloribates laevigatus was a marker of bare soil at a shallow depth and Vulgoramasus remberti of buried decomposition, not specific to any decay stage. Analysis of mite assemblages associated with head, torso and posterior body showed that Parasitus evertsi and M. matrius are attracted to beneath the thighs, whereas L. reticulata to beneath the head. This study highlights the value of mites as indicator species of decomposition and its stages, confirming (1) a succession of Acari on buried remains and (2) species specificity to body regions.

Microscopic analysis of the microbiota of three commercial Phytoseiidae species (Acari: Mesostigmata).

Microbes associated with the external and internal anatomy of three commercially available predatory mite species-Phytoseiulus persimilis, Typhlodromips (= Amblyseius) swirskii, and Neoseiulus (= Amblyseius) cucumeris-were examined using light microscopy, confocal laser scanning microscopy and fluorescence in-situ hybridization (FISH). Four microbe morphotypes were observed on external body regions. These included three microfungi-like organisms (named T1, T2 and T3) and rod-shaped bacteria (T4). Morphotypes showed unique distributions on the external body regions and certain microbes were found only on one host species. Microfungi-like T1 were present in all three species whereas T2 and T3 were present in only P. persimilis and T. swirskii, respectively. T1 and T2 microbes were most abundant on the ventral structures of the idiosoma and legs, most frequently associated with coxae, coxal folds, ventrianal shields and epigynal shields. T3 microbes were most abundant on legs and dorsal idiosoma. T4 microbes were less abundant and were attached to epigynal shields of N. cucumeris and T. swirskii. Significant differences in distribution between batches suggest temporal fluctuations in the microbiota of phytoseiids in mass-reared systems. FISH showed bacteria within the alimentary tract, in Malpighian tubules and anal atria. These may aid absorption of excretory products or maintaining gut physiology. We suggest a mechanism by which microbes may be transmitted to offspring and throughout populations. This study aims to improve our knowledge of this poorly understood area and highlights the necessity of understanding the microbiota of Acari.

Using human head lice to unravel neglect and cause of death.

Despite the common association of human lice with abandoned or neglected people, no procedure to assess pediculosis, aimed to detect signs of neglect, exists. Investigating the two most common forms of head louse infestation, regular and severe, we define lice-markers of neglect and develop a protocol and survey form to record and assess pediculosis. The study of head lice from a deceased victim of neglect helped unravel time-length since death, frequency of exposure to neglect and the cause and circumstances related to the death. Nit-clusters are markers of neglect, indicating length and frequency of neglect episodes. In the case study used here that culminated in the death of the victim, sustained abandonment started circa 2 years before discovery. The lice suggested that death was caused by overconsumption of a powerful calcium channel blocker, an antihypertensive, an excess of which in lice food supply (blood) stops oogenesis. Despite hosting thousands of adult females on the hair, lice reproduction stopped and nits were no longer developed or deposited on the hairs at the root end. This short distance of the shaft with no nits provided a time estimation of overdosing of almost 2 months before death.

Macrocheles species (Acari: Macrochelidae) associated with human corpses in Europe.

The biology of macrochelid mites might offer new venues for the interpretation of the environmental conditions surrounding human death and decomposition. Three human corpses, one from Sweden and two from Spain, have been analysed for the occurrence of Macrochelidae species. Macrocheles muscaedomesticae (Scopoli) females were associated with a corpse that was found in a popular beach area of southeast Spain. Their arrival coincides with the occurrence of one of their major carrier species, the filth fly Fannia scalaris, the activity of which peaks during mid-summer. Macrocheles glaber (Müller) specimens were collected from a corpse in a shallow grave in a forest in Sweden at the end of summer, concurrent with the arrival of beetles attracted by odours from the corpse. Macrocheles perglaber Filipponi and Pegazzano adults were sampled from a corpse found indoors in the rural surroundings of Granada city, south Spain. The phoretic behaviour of this species is similar to that of M. glaber, but it is more specific to Scarabaeidae and Geotrupidae dung beetles, most of which favour human faeces. Macrocheles muscaedomesticae is known from urban and rural areas and poultry farms, M. glaber from outdoors, particularly the countryside, whereas M. perglaber is known from outdoor, rural, and remote, potentially mountainous locations. Macrocheles muscaedomesticae and M. perglaber are reported for the first time from the Iberian Peninsula. This is the first record of M. perglaber from human remains.

Plasmin and coagulant activities in a minicurd model system: Study of technological parameters.

The effect of scalding temperature of the curd, the inclusion of a washing step, and the pH at whey drainage on plasmin and coagulant activities were assessed in a minicurd model of young hard cooked cheese. The variables were tested as follows: draining pH was assayed at 3 levels (4.6, 5.6, and 6.4), curd scalding temperature was tested at 50 and 56°C, and washing of the curd was examined at 2 levels (no washing step, and the replacement of the whey by water). Increase in pH at whey drainage and washing of the curd had a positive effect on plasmin activity, which was also evidenced by compatible changes in soluble peptide profiles. No effect of increased cooking temperature was found on plasmin activity. Plasminogen activation was not verified in any treatment. As for coagulant, lower pH values at whey drainage and a decrease in curd cooking temperature increased its activity; washing of the curd showed no influence on coagulant residual activity. These results were consistent with proteolysis described by peptide profiles, electrophoresis, and soluble nitrogen fractions.

Detection and localization of Solitalea-like and Cardinium bacteria in three Acarus siro populations (Astigmata: Acaridae).

Bacteria associated with mites influence their fitness, nutrition and reproduction. Previously, we found Solitalea-like (Sphingobacteriales) and Candidatus Cardinium (Cytophagales) bacteria in the stored product mite Acarus siro L. by cloning and using pyrosequencing. In this study, taxon-specific primers targeting 16S rRNA gene were used to detect and quantify the bacteria in mites and eggs of three A. siro populations. The specific probes for fluorescent in situ hybridization (FISH) were used to localize Solitalea-like and Cardinium bacteria in mite bodies. The population growth as an indirect estimator of fitness was used to describe the mite-bacteria interactions on (1) control diet; (2) rifampicin supplemented diet; (3) tetracycline supplemented diet; (4) rifampicin pretreated mites; (5) tetracycline pretreated mites. Solitalea-like 16S rRNA gene sequences from A. siro formed a separate cluster together with sequences from Tyrophagus putrescentiae. qPCR analysis indicated that number of Solitalea-like bacteria 16S rRNA gene copies was ca. 100× higher than that of Cardinium and the numbers differed between populations. FISH analysis localized Solitalea-like bacteria in the parenchymal tissues, mesodeum and food bolus of larvae, nymphs and adults. Solitalea-like, but not Cardinium bacteria were detected by taxon-specific primers in mites and eggs of all three investigated populations. None of the antibiotic treatments eliminated Solitalea-like bacteria in the A. siro populations tested. Rifampicin pretreatment significantly decreased the population growth. The numbers of Solitalea-like bacteria did not correlate with the population growth as a fitness indicator. This study demonstrated that A. siro can host Solitalea-like bacteria either alone or together with Cardinium. We suggest that Solitalea-like bacteria are shared by vertical transfer in A. siro populations.

Short communication: A new minicurd model system for hard cooked cheeses.

The aim of this study was to propose and validate a new minicurd model of young hard cheese. Curd particles and whey obtained from conventional cheese making of Reggianito Argentino were separated and frozen. Then, both fractions were thawed and the mixture of whey and curds was reconstituted, from which minicurds were made on the laboratory scale. Repeatability and the effect of freezing on minicurd composition were investigated by assessing pH, protein and moisture contents, sodium chloride content, and total thermophilic lactic flora counts. Good repeatability was achieved, and no significant differences were found between minicurds made from fresh compared with frozen materials. Composition of the minicurd was appropriate for modeling Reggianito Argentino cheese.

Ixodes ricinus (Ixodidae), an occasional phoront on necrophagous and coprophagous beetles in Europe.

For ticks, phoretic behaviour using insects associated with vertebrates might offer an alternative strategy to host-seeking. Here we report for the first time the presence of immature stages of the most widespread tick species in Western Europe, Ixodes ricinus (Acari: Ixodidae), on three beetle species belonging to families Silphidae and Geotrupidae (Coleoptera). Specimens were collected while performing fieldwork surveys on insect diversity during the peak of tick's questing behaviour, in July and August of 2009 and 2010. The collections took place in two Natural Parks, the Aiako Harria, Guipúzcoa in Northern Spain and Wellington Country Park, Berkshire, in England. The silphid beetle Nicrophorus vespilloides and the geotrupid Trypocopris pyrenaeus were collected from pig-carcasses and both carried nymphs of I. ricinus; the geotrupid Anoplotrupes stercorosus was carrying a tick larva while feeding on red deer dung. These findings revealed an unnoticed but common relation of ticks not only with decomposed animals but also with insect scavengers. We discuss the rationale of this phenomenon.

Genomic and metagenomic analyses of the domestic mite Tyrophagus putrescentiae identify it as a widespread environmental contaminant and a host of a basal, mite-specific Wolbachia lineage (supergroup Q).

Tyrophagus putrescentiae (mould mite) is a global, microscopic trophic generalist that commonly occurs in various human-created habitats, causing allergies and damaging stored food. Its ubiquity and extraordinary ability to penetrate research samples or cultures through air currents or by active walking through tights spaces (such as treads of screw caps) may lead to sample contamination and introduction of its DNA to research materials in the laboratory. This prompts a thorough investigation into potential sequence contamination in public genomic databases. The trophic success of T. putrescentiae is primarily attributed to the symbiotic bacteria housed in specialized internal mite structures, facilitating adaptation to varied nutritional niches. However, recent work suggests that horizontal transfer of bacterial/fungal genes related to nutritional functionality may also contribute to the mite's trophic versatility. This aspect requires independent confirmation. Additionally, T. putrescentiae harbors an uncharacterized and genetically divergent bacterium, Wolbachia, displaying blocking and microbiome-modifying effects. The phylogenomic position and supergroup assignment of this bacterium are unknown. Here, we sequenced and assembled the T. putrescentiae genome, analyzed its microbiome, and performed detailed phylogenomic analyses of the mite-specific Wolbachia. We show that T. putrescentiae DNA is a substantial source of contamination of research samples. Its DNA may inadvertently be co-extracted with the DNA of the target organism, eventually leading to sequence contamination in public databases. We identified a diversity of bacterial species associated with T. putrescentiae, including those capable of rapidly developing antibiotic resistance, such as Escherichia coli. Despite the presence of diverse bacterial communities in T. putrescentiae, we did not detect any recent horizontal gene transfers in this mite species and/or in astigmatid (domestic) mites in general. Our phylogenomic analysis of Wolbachia recovered a basal, mite-specific lineage (supergroup Q) represented by two Wolbachia spp. from the mould mite and a gall-inducing plant mite. Fluorescence in situ hybridization confirmed the presence of Wolbachia inside the mould mite. The discovery of an early derivative Wolbachia lineage (supergroup Q) in two phylogenetically unrelated and ecologically dissimilar mites suggests that this endosymbiotic bacterial lineage formed a long-term association with mites. This finding provides a unique insight into the early evolution and host associations of Wolbachia. Further discoveries of Wolbachia diversity in acariform mites are anticipated.

Specific detection and localization of microsporidian parasites in invertebrate hosts by using in situ hybridization.

We designed fluorescence in situ hybridization probes for two distinct microsporidian clades and demonstrated their application in detecting, respectively, Nosema/Vairimorpha and Dictyoceola species. We used them to study the vertical transmission of two microsporidia infecting the amphipod Gammarus duebeni.

Body fat mass is a strong and negative predictor of peak stimulated growth hormone and bone mineral density in healthy adolescents during transition period.

The adipose tissue has detrimental effects on growth hormone secretion, even in the absence of obesity. The majority of previous studies have shown an inverse relationship between fat mass and the growth hormone response to several stimulation tests in adults and in children. The contribution of body fat mass on growth hormone response to provocative tests during the transition age is not known. We analyzed the GH-IGF1 axis by GHRH-arginine test in 30 healthy adolescents with normal stature during the transition period from 14 to 18 years. All subjects underwent body composition analysis by dual-energy X-ray absorptiometry (DXA). We found that total body fat mass was inversely correlated with peak GH to provocative test (r=-0.6, p=0.004). GH deficiency was shown in 2 of our healthy patients if diagnosis was based on GH peak below 19 µg/l. Both children who failed the GHRH-arginine were overweight (BMI for age above 85th percentile). However, their GH status was normal when assessed by insulin tolerance test. Multivariate analysis demonstrated strong correlation between peak stimulated GH and measures of body adiposity, including body mass index and fat mass index, with the latter showing the most important effect on GH secretion. Fat mass index alone explained 34.5% of the variability in peak GH. This study has shown for the first time that during the transition period, GH response to GHRH-arginine test is strongly influenced by body composition, and cutoff values appropriate for overweight and obese adolescents are needed.

Chasing the Patagonian sun: comparative thermal biology of Liolaemus lizards.

The importance of the thermal environment for ectotherms and its relationship with thermal physiology and ecology is widely recognized. Several models have been proposed to explain the evolution of the thermal biology of ectotherms, but experimental studies have provided mixed support. Lizards from the Liolaemus goetschi group can be found along a wide latitudinal range across Argentina. The group is monophyletic and widely distributed, and therefore provides excellent opportunities to study the evolution of thermal biology. We studied thermal variables of 13 species of the L. goetschi group, in order to answer three questions. First, are aspects of the thermal biology of the L. goetschi group modelled by the environment or are they evolutionarily conservative? Second, have thermal characteristics of these animals co-evolved? And third, how do the patterns of co-evolution observed within the L. goetschi group compare to those in a taxonomically wider selection of species of Liolaemus? We collected data on 13 focal species and used species information of Liolaemus lizards available in the literature and additional data obtained by the authors. We tackled these questions using both conventional and phylogenetically based analyses. Our results show that lizards from the L. goetschi group and the genus Liolaemus in general vary in critical thermal minimum in relation to mean air temperature, and particularly the L. goetschi group shows that air temperature is associated with critical thermal range, as well as with body temperature. Although the effect of phylogeny cannot be ignored, our results indicate that these thermal biology aspects are modelled by cold environments of Patagonia, while other aspects (preferred body temperature and critical thermal maximum) are more conservative. We found evidence of co-evolutionary patterns between critical thermal minimum and preferred body temperature at both phylogenetic scales (the L. goetschi group and the extended sample of 68 Liolaemus species).

Cardinium endosymbionts are widespread in synanthropic mite species (Acari: Astigmata).

'Candidatus Cardinium' is an intracellular endosymbiont or parasite frequently occurring in invertebrates including mites and ticks. In this work we report Cardinium bacteria in Astigmata mites and explore their incidence in synanthropic species. Amplification of a 776 bp bacterial 16S rRNA gene fragment, using specific primers, enabled identification of closely related Cardinium sequences in 13 laboratory-reared populations of mites. In addition, Cardinium sequences were identified in three wild mite populations. Large scale screening of these populations showed 100% prevalence of Cardinium, representing the highest incidence compared to other major Chelicerate groups.

Genome sequences of the human body louse and its primary endosymbiont provide insights into the permanent parasitic lifestyle.

As an obligatory parasite of humans, the body louse (Pediculus humanus humanus) is an important vector for human diseases, including epidemic typhus, relapsing fever, and trench fever. Here, we present genome sequences of the body louse and its primary bacterial endosymbiont Candidatus Riesia pediculicola. The body louse has the smallest known insect genome, spanning 108 Mb. Despite its status as an obligate parasite, it retains a remarkably complete basal insect repertoire of 10,773 protein-coding genes and 57 microRNAs. Representing hemimetabolous insects, the genome of the body louse thus provides a reference for studies of holometabolous insects. Compared with other insect genomes, the body louse genome contains significantly fewer genes associated with environmental sensing and response, including odorant and gustatory receptors and detoxifying enzymes. The unique architecture of the 18 minicircular mitochondrial chromosomes of the body louse may be linked to the loss of the gene encoding the mitochondrial single-stranded DNA binding protein. The genome of the obligatory louse endosymbiont Candidatus Riesia pediculicola encodes less than 600 genes on a short, linear chromosome and a circular plasmid. The plasmid harbors a unique arrangement of genes required for the synthesis of pantothenate, an essential vitamin deficient in the louse diet. The human body louse, its primary endosymbiont, and the bacterial pathogens that it vectors all possess genomes reduced in size compared with their free-living close relatives. Thus, the body louse genome project offers unique information and tools to use in advancing understanding of coevolution among vectors, symbionts, and pathogens.

Occurrence of Poecilochirus austroasiaticus (Acari: Parasitidae) in forensic autopsies and its application on postmortem interval estimation.

Despite the fact that mites were used at the dawn of forensic entomology to elucidate the postmortem interval, their use in current cases remains quite low for procedural reasons such as inadequate taxonomic knowledge. A special interest is focused on the phoretic stages of some mite species, because the phoront-host specificity allows us to deduce in many occasions the presence of the carrier (usually Diptera or Coleoptera) although it has not been seen in the sampling performed in situ or in the autopsy room. In this article, we describe two cases where Poecilochirus austroasiaticus Vitzthum (Acari: Parasitidae) was sampled in the autopsy room. In the first case, we could sample the host, Thanatophilus ruficornis (Küster) (Coleoptera: Silphidae), which was still carrying phoretic stages of the mite on the body. That attachment allowed, by observing starvation/feeding periods as a function of the digestive tract filling, the establishment of chronological cycles of phoretic behavior, showing maximum peaks of phoronts during arrival and departure from the corpse and the lowest values in the phase of host feeding. From the sarcosaprophagous fauna, we were able to determine in this case a minimum postmortem interval of 10 days. In the second case, we found no Silphidae at the place where the corpse was found or at the autopsy, but a postmortem interval of 13 days could be established by the high specificity of this interspecific relationship and the departure from the corpse of this family of Coleoptera.