Glycogen-autophagy: Molecular machinery and cellular mechanisms of glycophagy.

Autophagy is an essential cellular process involving degradation of superfluous or defective macromolecules and organelles as a form of homeostatic recycling. Initially proposed to be a "bulk" degradation pathway, a more nuanced appreciation of selective autophagy pathways has developed in the literature in recent years. As a glycogen-selective autophagy process, "glycophagy" is emerging as a key metabolic route of transport and delivery of glycolytic fuel substrate. Study of glycophagy is at an early stage. Enhanced understanding of this major noncanonical pathway of glycogen flux will provide important opportunities for new insights into cellular energy metabolism. In addition, glycogen metabolic mishandling is centrally involved in the pathophysiology of several metabolic diseases in a wide range of tissues, including the liver, skeletal muscle, cardiac muscle, and brain. Thus, advances in this exciting new field are of broad multidisciplinary interest relevant to many cell types and metabolic states. Here, we review the current evidence of glycophagy involvement in homeostatic cellular metabolic processes and of molecular mediators participating in glycophagy flux. We integrate information from a variety of settings including cell lines, primary cell culture systems, ex vivo tissue preparations, genetic disease models, and clinical glycogen disease states.

Cost-effectiveness of cenobamate for focal seizures in people with drug-resistant epilepsy.

OBJECTIVE: This study was undertaken to estimate the cost-effectiveness of add-on cenobamate in the UK when used to treat drug-resistant focal seizures in adults who are not adequately controlled with at least two prior antiseizure medications, including at least one used adjunctively. METHODS: We estimated the cost per quality-adjusted life-year (QALY) for cenobamate compared to brivaracetam, eslicarbazepine, lacosamide, and perampanel in the UK National Health Service over a lifetime time horizon. We used a Markov cohort structure to determine response to treatment, using pooled data from three long-term studies of cenobamate. A network meta-analysis informed the likelihood of response to therapy with brivaracetam, eslicarbazepine, lacosamide, and perampanel relative to cenobamate. Once individuals discontinued treatment, they transitioned to subsequent treatment health states, including other antiseizure medicines, surgery, and vagus nerve stimulation. Costs included treatment, administration, routine monitoring, event management, and adverse events. Published evidence and expert opinion informed the likelihood of response to subsequent treatments, associated adverse events, and costs. Utility data were based on Short-Form six-dimension form utility. Discounting was applied at 3.5% per annum as per National Institute for Health and Care Excellence guidance. Uncertainty was explored through deterministic and probabilistic sensitivity analyses. RESULTS: In the base case, cenobamate led to cost savings of £51 967 (compared to brivaracetam), £21 080 (compared to eslicarbazepine), £33 619 (compared to lacosamide), and £28 296 (compared to perampanel) and increased QALYs of 1.047 (compared to brivaracetam), 0.598 (compared to eslicarbazepine), 0.776 (compared to lacosamide), and 0.703 (compared to perampanel) per individual over a lifetime time horizon. Cenobamate also dominated the four drugs across most sensitivity analyses. Differences were due to reduced seizure frequency with cenobamate relative to comparators. SIGNIFICANCE: Cenobamate improved QALYs and was less costly than brivaracetam, eslicarbazepine, lacosamide, and perampanel. Therefore, cenobamate may be considered as a cost-effective adjunctive antiseizure medication for people with drug-resistant focal seizures.

α1-Antitrypsin A treatment attenuates neutrophil elastase accumulation and enhances insulin sensitivity in adipose tissue of mice fed a high-fat diet.

Neutrophils accumulate in insulin-sensitive tissues during obesity and may play a role in impairing insulin sensitivity. The major serine protease expressed by neutrophils is neutrophil elastase (NE), which is inhibited endogenously by α1-antitrypsin A (A1AT). We investigated the effect of exogenous (A1AT) treatment on diet-induced metabolic dysfunction. Male C57Bl/6j mice fed a chow or a high-fat diet (HFD) were randomized to receive intraperitoneal injections three times weekly of either Prolastin (human A1AT; 2 mg) or vehicle (PBS) for 10 wk. Prolastin treatment did not affect plasma NE concentration, body weight, glucose tolerance, or insulin sensitivity in chow-fed mice. In contrast, Prolastin treatment attenuated HFD-induced increases in plasma and white adipose tissue (WAT) NE without affecting circulatory neutrophil levels or increases in body weight. Prolastin-treated mice fed a HFD had improved insulin sensitivity, as assessed by insulin tolerance test, and this was associated with higher insulin-dependent IRS-1 (insulin receptor substrate) and AktSer473 phosphorylation, and reduced inflammation markers in WAT but not liver or muscle. In 3T3-L1 adipocytes, Prolastin reversed recombinant NE-induced impairment of insulin-stimulated glucose uptake and IRS-1 phosphorylation. Furthermore, PDGF mediated p-AktSer473 activation and glucose uptake (which is independent of IRS-1) was not affected by recombinant NE treatment. Collectively, our findings suggest that NE infiltration of WAT during metabolic overload contributes to insulin resistance by impairing insulin-induced IRS-1 signaling.NEW & NOTEWORTHY Neutrophils accumulate in peripheral tissues during obesity and are critical coordinators of tissue inflammatory responses. Here, we provide evidence that inhibition of the primary neutrophil protease, neutrophil elastase, with α1-antitrypsin A (A1AT) can improve insulin sensitivity and glucose homeostasis of mice fed a high-fat diet. This was attributed to improved insulin-induced IRS-1 phosphorylation in white adipose tissue and provides further support for a role of neutrophils in mediating diet-induced peripheral tissue insulin resistance.

Probabilistic seasonal dengue forecasting in Vietnam: A modelling study using superensembles.

BACKGROUND: With enough advanced notice, dengue outbreaks can be mitigated. As a climate-sensitive disease, environmental conditions and past patterns of dengue can be used to make predictions about future outbreak risk. These predictions improve public health planning and decision-making to ultimately reduce the burden of disease. Past approaches to dengue forecasting have used seasonal climate forecasts, but the predictive ability of a system using different lead times in a year-round prediction system has been seldom explored. Moreover, the transition from theoretical to operational systems integrated with disease control activities is rare. METHODS AND FINDINGS: We introduce an operational seasonal dengue forecasting system for Vietnam where Earth observations, seasonal climate forecasts, and lagged dengue cases are used to drive a superensemble of probabilistic dengue models to predict dengue risk up to 6 months ahead. Bayesian spatiotemporal models were fit to 19 years (2002-2020) of dengue data at the province level across Vietnam. A superensemble of these models then makes probabilistic predictions of dengue incidence at various future time points aligned with key Vietnamese decision and planning deadlines. We demonstrate that the superensemble generates more accurate predictions of dengue incidence than the individual models it incorporates across a suite of time horizons and transmission settings. Using historical data, the superensemble made slightly more accurate predictions (continuous rank probability score [CRPS] = 66.8, 95% CI 60.6-148.0) than a baseline model which forecasts the same incidence rate every month (CRPS = 79.4, 95% CI 78.5-80.5) at lead times of 1 to 3 months, albeit with larger uncertainty. The outbreak detection capability of the superensemble was considerably larger (69%) than that of the baseline model (54.5%). Predictions were most accurate in southern Vietnam, an area that experiences semi-regular seasonal dengue transmission. The system also demonstrated added value across multiple areas compared to previous practice of not using a forecast. We use the system to make a prospective prediction for dengue incidence in Vietnam for the period May to October 2020. Prospective predictions made with the superensemble were slightly more accurate (CRPS = 110, 95% CI 102-575) than those made with the baseline model (CRPS = 125, 95% CI 120-168) but had larger uncertainty. Finally, we propose a framework for the evaluation of probabilistic predictions. Despite the demonstrated value of our forecasting system, the approach is limited by the consistency of the dengue case data, as well as the lack of publicly available, continuous, and long-term data sets on mosquito control efforts and serotype-specific case data. CONCLUSIONS: This study shows that by combining detailed Earth observation data, seasonal climate forecasts, and state-of-the-art models, dengue outbreaks can be predicted across a broad range of settings, with enough lead time to meaningfully inform dengue control. While our system omits some important variables not currently available at a subnational scale, the majority of past outbreaks could be predicted up to 3 months ahead. Over the next 2 years, the system will be prospectively evaluated and, if successful, potentially extended to other areas and other climate-sensitive disease systems.

A "Dirty" Footprint: Macroinvertebrate diversity in Amazonian Anthropic Soils.

Amazonian rainforests, once thought to be pristine wilderness, are increasingly known to have been widely inhabited, modified, and managed prior to European arrival, by human populations with diverse cultural backgrounds. Amazonian Dark Earths (ADEs) are fertile soils found throughout the Amazon Basin, created by pre-Columbian societies with sedentary habits. Much is known about the chemistry of these soils, yet their zoology has been neglected. Hence, we characterized soil fertility, macroinvertebrate communities, and their activity at nine archeological sites in three Amazonian regions in ADEs and adjacent reference soils under native forest (young and old) and agricultural systems. We found 673 morphospecies and, despite similar richness in ADEs (385 spp.) and reference soils (399 spp.), we identified a tenacious pre-Columbian footprint, with 49% of morphospecies found exclusively in ADEs. Termite and total macroinvertebrate abundance were higher in reference soils, while soil fertility and macroinvertebrate activity were higher in the ADEs, and associated with larger earthworm quantities and biomass. We show that ADE habitats have a unique pool of species, but that modern land use of ADEs decreases their populations, diversity, and contributions to soil functioning. These findings support the idea that humans created and sustained high-fertility ecosystems that persist today, altering biodiversity patterns in Amazonia.

Phylogenomic analyses of Crassiclitellata support major Northern and Southern Hemisphere clades and a Pangaean origin for earthworms.

BACKGROUND: Earthworms (Crassiclitellata) are a diverse group of annelids of substantial ecological and economic importance. Earthworms are primarily terrestrial infaunal animals, and as such are probably rather poor natural dispersers. Therefore, the near global distribution of earthworms reflects an old and likely complex evolutionary history. Despite a long-standing interest in Crassiclitellata, relationships among and within major clades remain unresolved. METHODS: In this study, we evaluate crassiclitellate phylogenetic relationships using 38 new transcriptomes in combination with publicly available transcriptome data. Our data include representatives of nearly all extant earthworm families and a representative of Moniligastridae, another terrestrial annelid group thought to be closely related to Crassiclitellata. We use a series of differentially filtered data matrices and analyses to examine the effects of data partitioning, missing data, compositional and branch-length heterogeneity, and outgroup inclusion. RESULTS AND DISCUSSION: We recover a consistent, strongly supported ingroup topology irrespective of differences in methodology. The topology supports two major earthworm clades, each of which consists of a Northern Hemisphere subclade and a Southern Hemisphere subclade. Divergence time analysis results are concordant with the hypothesis that these north-south splits are the result of the breakup of the supercontinent Pangaea. CONCLUSIONS: These results support several recently proposed revisions to the classical understanding of earthworm phylogeny, reveal two major clades that seem to reflect Pangaean distributions, and raise new questions about earthworm evolutionary relationships.

Phylogenetic evaluation of Amynthas earthworms from South China reveals the initial ancestral state of spermathecae.

Our knowledge of the phylogeny of the earthworm genus Amynthas under the family Megascolecidae, which is comprised of a huge number of species, is very limited compared to the better-known and much smaller family Lumbricidae. In order to investigate the phylogenetic relationships among the species within the genus Amynthas, which is the largest genus of the Megascolecidae family, nuclear and mitochondrial DNA sequences of 77 species, including 76 in-group Amynthas species collected from South China and 1 out-group species, were analyzed. A 5402bp segments composed of whole nuclear 18S rDNA and the mitochondrial genes COI, COII, ND1, 12S, and 16S was assembled from 77 species. Maximum Likelihood and Bayesian analyses of the concatenated sequences were performed. The results revealed evolution of two geographically independent lineages, both showing the ancestral state of two pairs of spermatheca (Sp.p 7/8/9). We found the species groups described by Sims and Easton (1972) to be non-monophyletic, and the origin of the parthenogenetic species group to likely be a quadthecal ancestor. These results provide modest evidence in support of an Indochinese peninsula origin of the Chinese Amynthas species and divergence of the genus once it had spread to mainland China. The findings of this study are consistent with a divergence scenario that resulted in at least one branch spreading to the Southeast of China and another branch spreading to the Southwest of China, but further research is required to confirm this interpretation of the Amynthas phylogeny.

Underground evolution: new roots for the old tree of lumbricid earthworms.

Earthworms belonging to the family Lumbricidae are extremely abundant in terrestrial temperate regions. They affect soil properties and nutrient cycling, thus shaping plant community composition and aboveground food webs. Some lumbricids are also model organisms in ecology and toxicology. Despite the intense research efforts dedicated to lumbricids over the last 130years, the evolutionary relationships and taxonomic classification of these organisms are still subject to great debate. Resolution of their systematics is hampered by the structural simplicity of the earthworm body plan and the existence of cryptic species. We sampled 160 earthworm specimens belonging to 84 lumbricid species (28 genera) and 22 Lumbricoidea outgroups, sequenced two nuclear genes, four mitochondrial genes and seven mitochondrial tRNAs and examined 22 morphological characters. We then applied a combination of phylogenetic methods to generate the first robust genus-level phylogeny of the Lumbricidae. Our results show that the current Lumbricidae classification and the underlying hypotheses of character evolution must be revised. Our chronogram suggests that lumbricids emerged in the Lower Cretaceous in the holarctic region and that their diversification has been driven by tectonic processes (e.g. Laurasia split) and geographical isolation. Our chronogram and character reconstruction analysis reveal that spermathecae number does not follow a gradual pattern of reduction and that parthenogenesis arose from sexual relatives multiple times in the group; the same analysis also indicates that both epigeic and anecic earthworms evolved from endogeic ancestors. These findings emphasize the strong and multiple changes to which morphological and ecological characters are subjected, challenging the hypothesis of character stasis in Lumbricidae.

Three new earthworms of the Amynthas corticis-group (Clitellata: Megascolecidae) from the Nam Et-Phouleoi, Laos.

Three new earthworms of the Amynthas corticis-group (Clitellata: Megascolecidae) from Phouleoi Mountain, Lao PDR. Earthworm specimens were collected from Mt. Phouleoi, Viengthong District, Houaphanh Province, Lao PDR. The species belong to the pheretimoid earthworms. All three holandric species key out to the A. corticis-group in Sims & Easton (1972), having spermathecal pores in 5/68/9. The new species are: Amynthas ammalinei sp. nov., Amynthas xayi sp. nov., and Amynthas piyakarnae sp. nov. Amynthas ammalinei sp. nov. has male pores superficial on small round porophores, 0.160.31 circumference apart. Amynthas xayi sp. nov. has male pores superficial in XVIII at centers of round porophores, 0.170.29 circumference apart. Amynthas piyakarnae sp. nov. has male pores superficial on transverse elevated elongate oval pads equatorial on XVIII, pads surrounded by 34 small outer rings, 0.250.31 circumference apart.

Correction: Linking bacterial enterotoxins and alpha defensin 5 expansion in the Crohn's colitis: A new insight into the etiopathogenetic and differentiation triggers driving colonic inflammatory bowel disease.

[This corrects the article DOI: 10.1371/journal.pone.0246393.].

Soil macrofauna communities in Brazilian land-use systems.

Background: Soil animal communities include more than 40 higher-order taxa, representing over 23% of all described species. These animals have a wide range of feeding sources and contribute to several important soil functions and ecosystem services. Although many studies have assessed macroinvertebrate communities in Brazil, few of them have been published in journals and even fewer have made the data openly available for consultation and further use. As part of ongoing efforts to synthesise the global soil macrofauna communities and to increase the amount of openly-accessible data in GBIF and other repositories related to soil biodiversity, the present paper provides links to 29 soil macroinvertebrate datasets covering 42 soil fauna taxa, collected in various land-use systems in Brazil. A total of 83,085 georeferenced occurrences of these taxa are presented, based on quantitative estimates performed using a standardised sampling method commonly adopted worldwide to collect soil macrofauna populations, i.e. the TSBF (Tropical Soil Biology and Fertility Programme) protocol. This consists of digging soil monoliths of 25 x 25 cm area, with handsorting of the macroinvertebrates visible to the naked eye from the surface litter and from within the soil, typically in the upper 0-20 cm layer (but sometimes shallower, i.e. top 0-10 cm or deeper to 0-40 cm, depending on the site). The land-use systems included anthropogenic sites managed with agricultural systems (e.g. pastures, annual and perennial crops, agroforestry), as well as planted forests and native vegetation located mostly in the southern Brazilian State of Paraná (96 sites), with a few additional sites in the neighbouring states of São Paulo (21 sites) and Santa Catarina (five sites). Important metadata on soil properties, particularly soil chemical parameters (mainly pH, C, P, Ca, K, Mg, Al contents, exchangeable acidity, Cation Exchange Capacity, Base Saturation and, infrequently, total N), particle size distribution (mainly % sand, silt and clay) and, infrequently, soil moisture and bulk density, as well as on human management practices (land use and vegetation cover) are provided. These data will be particularly useful for those interested in estimating land-use change impacts on soil biodiversity and its implications for below-ground foodwebs, ecosystem functioning and ecosystem service delivery. New information: Quantitative estimates are provided for 42 soil animal taxa, for two biodiversity hotspots: the Brazilian Atlantic Forest and Cerrado biomes. Data are provided at the individual monolith level, representing sampling events ranging from February 2001 up to September 2016 in 122 sampling sites and over 1800 samples, for a total of 83,085 ocurrences.

Coordinate patterns of estrogen receptor, progesterone receptor, and Wilms tumor 1 expression in the histopathologic distinction of ovarian from endometrial serous adenocarcinomas.

The purpose of this study is to assess whether composite or coordinate immunoexpression patterns of estrogen receptor (ER), progesterone receptor (PR), and Wilms tumor 1 (WT1) gene can significantly distinguish between endometrial serous carcinoma (ESC) and ovarian serous carcinoma (OSC). Immunohistochemical analyses were performed on whole tissue sections from 22 uterus-confined ESCs and on a tissue microarray of 140 high-grade, pan-stage OSCs, using antibodies to ER, PR, and WT-1. Estrogen receptor, PR, and WT1 expressions were present in 37%, 49%, and 81% of OSC, respectively, but these markers were also present in 18%, 27%, and 36% of ESC. The ER+/PR+/WT1+ coordinate profile was identified in 33.6% of OSC but in none of ESC (P = .0006), resulting in a calculated sensitivity and specificity of this profile for OSC of 33.6% and 100%, respectively. By contrast, the ER-/PR-/WT1- coordinate profile was identified in 41% of ESC but in only 6.4% of OSC (P = .0001), resulting in a calculated sensitivity and specificity of this profile for ESC of 50% and 94%. In summary, in the differential diagnosis between OSC and ESC, positivity for all 3 markers favors an extrauterine origin, whereas negativity for all 3 markers is supportive of an endometrial origin. The use of single markers for this purpose is not recommended, as each lacks optimal discriminatory power. Coordinate profiles, in general, have a high specificity but low sensitivity in this differential diagnosis.

Three new earthworm species of the genus Amynthas (Clitellata: Megascolecidae) from Mt. Chiak National Park, Korea.

Earthworm specimens collected from Mt. Chiak, National Park, Korea were found to represent three new species of megascolecid earthworms: Amynthas chiakensis sp. nov., Amynthas gyeongriae sp. nov., and Amynthas wonjuensis sp. nov. Amynthas chiakensis sp. nov. has two pairs of spermathecae in VII and VIII, small transverse oval male porophores each with C-shaped small patches of genital papillae, 0.22-0.31 circumference apart. Amynthas gyeongriae sp. nov. has spermathecae in VI-VIII, male field with large circular-shaped raised pads and seminal grooves, with pores 0.16-0.28 circumference apart. Amynthas wonjuensis sp. nov. has spermathecae in VI and VII, male pores superficial in XVIII on small oval-shaped white porophores lateral to large roughly circular raised genital papillae, 0.15 circumference apart.

New Ocnerodrilidae genera, species and records from Brazil (Annelida: Crassiclitellata).

We describe three new species of Eukerria, six of Kerriona, one of Ocnerodrilus and two of Belladrilus (Clitellata: Ocnerodrilidae) from Brazil, mostly from wet habitats including bromeliad leaf axils, plus some from forest soils in Bahia state. We also provide new morphological and distribution data on Kerriona luederwaldti Michaelsen, 1924. We describe the new genus Pauqueba and its two new species, both from bromeliad leaf axils and forest floor litter in the state of Bahia. It has two pairs of calciferous glands, the balantine reduction of the male terminalia but spermathecae in ix, and no gizzard. The new genus Parabauba found in deeper (30 cm) soil layers is distinctive for the combination of two pairs of calciferous glands, a large gizzard in v, acanthodriline male terminalia, and no typholosole. The new genus Diplomoela from a mesic soil in Bahia is meronephric, unusually large for the Ocnerodrilidae, and has two well-developed gizzards.

Mitigating Mismatch Compression in Differential Local Field Potentials.

Deep brain stimulation (DBS) devices capable of measuring differential local field potentials ( ∂ LFP) enable neural recordings alongside clinical therapy. Efforts to identify oscillatory correlates of various brain disorders, or disease readouts, are growing but must proceed carefully to ensure readouts are not distorted by brain environment. In this report we identified, characterized, and mitigated a major source of distortion in ∂ LFP that we introduce as mismatch compression (MC). Using in vivo, in silico, and in vitro models of MC, we showed that impedance mismatches in the two recording electrodes can yield incomplete rejection of stimulation artifact and subsequent gain compression that distorts oscillatory power. We then developed and validated an opensource mitigation pipeline that mitigates the distortions arising from MC. This work enables more reliable oscillatory readouts for adaptive DBS applications.

Distribution and diversity of earthworms in different land use systems in Rio Grande do Sul, Brazil.

The southernmost state of Brazil, Rio Grande do Sul, includes both the Pampa and Atlantic Forest biomes, and covers an area of around 284 thousand km2. Since the last checklist for the state, published in 2012, many additional sites have been sampled. Hence, we provide here a full list of earthworm species and their distribution in different municipalities and land use systems in the state. Data were obtained from the literature, museums, and other earthworm collections and recent sampling efforts. Records for 19 additional species are reported here for the first time, raising the total to 78 species/subspecies currently known from Rio Grande do Sul, from the families Acanthodrilidae (5), Benhamiidae (3), Lumbricidae (13), Megascolecidae (11), Ocnerodrilidae (22), Almidae (2), Glossoscolecidae (18) and Rhinodrilidae (5). Alexidrilus Righi, 1971 is synonymized with Urobenus Benham, 1886, as both species in the genus (A. littoralis Ljungström, 1972 and A. lourdesae Righi, 1971) have similar morphology to the type species Urobenus brasiliensis Benham, 1886. Most of the native species/subspecies in the state (39), including an important number of new species, were found in native vegetation with lesser disturbance, such as swamps, native forests and pastures. On the other hand, the three peregrine and 27 exotic species (especially Pontoscolex corethrurus, Eukerria spp., lumbricids, megascolecids, Dichogaster spp., and Microscolex spp.) dominated in disturbed sites, such as compost piles, urban areas, rice and other crops, and cultivated pastures. Considering the geographic extension and the many unsampled sites in the state, further collection efforts are necessary and will certainly reveal new species, as evidenced here.

Earthworm species in different land use systems in the state of Goiás and the Federal District of Brazil.

The state of Goiás and the Federal District together cover an area of about 346 thousand km2, including the diversified Cerrado vegetation, also known as the Brazilian Savanna, a biodiversity hotspot with many endangered and endemic species. Much of the Cerrado has been converted to agriculture, but little is known of its earthworms, and the impacts of land use management and agricultural practices on their density and diversity. In the present paper we review the known earthworm species in Goiás and the Federal District based on historical and museum records, and update information on their distribution and populations in different counties and land use systems with data from recent samplings. Five additional species are reported here for the first time, raising the total to 12 species currently known from Goiás, from the families Benhamiidae (1), Glossoscolecidae (1), Megascolecidae (3), Ocnerodrilidae (2) and Rhinodrilidae (5). In the Federal District, an additional 10 species are reported, bringing the total to 23 earthworm species, belonging to the families Benhamiidae (4), Eudrilidae (1), Glossoscolecidae (2), Ocnerodrilidae (1), Lumbricidae (2), Megascolecidae (1) and Rhinodrilidae (12). Native species in Goiás (6) were found in areas with lesser disturbance, such as in native forest and flooded areas/river banks, a phenomenon also observed in the Federal District. On the other hand, the exotic and peregrine species found in Goiás (6) and in the Federal District (9) dominated in disturbed sites, such as perennial crops, agroforestry, annual crops, urban areas, secondary forests, pastures and agropastoral systems, among others. Given the many new species found and the extension of Goiás and the Federal District, further effort is needed to adequately characterize the earthworm fauna of the region, from where many other species are expected.

Earthworms from natural and managed ecosystems in Southern Bahia, Brazil.

The earthworms (Crassiclitellata) of the Northern Atlantic Forest region are poorly known, and many new species are expected to be found in the region. Hence, quantitative and qualitative samples were taken in the Atlantic Forest region of southern Bahia State, Brazil, at eight sites to assess species presence and abundance in different land uses (forests, pastures, plantations). Earthworms were also qualitatively collected at an additional eight sites. Only one to two species per site were found in quantitative samples, while qualitative samples generally resulted in higher species recoveries, with highest richness observed at the native Atlantic Forest of Veracel in Porto Seguro. Pontoscolex corethrurus (Müller, 1857) was the dominant species at all sites and in most land use systems, probably favored by human migration and agricultural activities; it was only absent in one primary forest site where native Ocnerodrilidae species predominated. In total at least 14 species were found of which eight were new, belonging to the genus Rhinodrilus and three new genera in the Ocnerodrilidae family (Pauqueba, Parabauba and Diplomoela). Contrary to sites in Southern and Southeastern Brazil, where many native species belong to the Glossoscolecidae family, in southern Bahia, Ocnerodrilidae species tend to be more prevalent, occupying various habitats. More intensive sampling of these habitats, including surface litter, in and under fallen logs and dead tree trunks, under rocks, in bromeliad leaf tanks and deeper soil layers is essential in order to properly characterize earthworm communities in Brazilian Atlantic Forests.

Ovary organization and ultrastructure in six species of Amynthas and Metaphire earthworms (Annelida, Crassiclitellata, Megascolecidae).

Ovaries in earthworms belonging to the family Megascolecidae are paired structures attached to the septum in the anterior part of the XIII segment. They are fan to rosette shaped with numerous rows of growing oocytes, known as egg strings, radiating from the ovary center towards the segmental cavity. The histological and ultrastructural ovary organization in megascolecids and the course of oogenesis remain unknown. The paper presents the results of light and electron microscopy analyses of ovaries in six megascolecid species, three from the genus Amynthas and three from Metaphire. Both parthenogenetic and sexually reproducing species were included in the study. The organization and ultrastructure of ovaries in all studied species are broadly similar. Considering the histological organization of ovaries, they could be divided into two zones. Zone I (proximal, close to the connection with the septum) is tightly packed with germline and somatic cells. Germ cells are interconnected via intercellular bridges and thin strands of the central cytoplasm (known as cytophore) and form syncytial cysts. Cysts unite oogonia, early meiotic cells (till diplotene), and clustering cells develop synchronously. During diplotene, interconnected cells lose developmental synchrony; most probably, one cell per cyst grows faster than others, detaches from the cysts, and becomes an oocyte. The remaining cells grow slightly and are still interconnected via the thin and reticular cytophore; these cells are considered nurse cells. Zone II has a form of egg strings where growing oocytes are isolated one from another by thin somatic cells and form short cords. We present the ultrastructural details of germline and somatic cells. We propose the term "Amynthas" type of ovaries for this ovary organization. We suppose that such ovaries are characteristic of other megascolecids and related families.