Neyraiella distinctus n. sp. (Oxyurida,Blattophilidae) parasite of nymphs of Gryllodes laplatae Sauss (Orthoptera,Gryllidae) in Argentina

Neyraiella distinctus n. sp. was found parasitizing nymphs of the cricket Gryllodes laplatae Sauss in City Bell, Argentina. This species was characterized by having the excretory pore in the posterior end of the basal bulb, vulva protruding with one lip well developed in the 1/3 end of the body, anus of the female with wings, male with a single spicule without any sculpture, gubernaculum and bursa are absent, six pairs of genital papillae arranged in two preanal pairs, one adanal pair and three postanal pairs, and the tail appendage in both sexes was short and conic.     

Tight steric closure at the intracellular activation gate of a voltage-gated K(+) channel.

In voltage-gated K(+) channels (Kv), an intracellular gate regulates access from the cytoplasm to the pore by organic channel blockers and by chemical modifiers. But is ion flow itself controlled instead by constriction of the narrow selectivity filter near the extracellular surface? We find that the intracellular gate of Kv channels is capable of regulating access even by the small cations Cd(2+) and Ag(+). It can also exclude small neutral or negatively charged molecules, indicating that the gate operates by steric exclusion rather than electrostatically. Just intracellular to the gated region, channel closure does not restrict access even to very large reagents. Either these Kv channels have a broader inner entrance than seen in the KcsA crystal, even in the closed state, or the region is highly flexible (but nevertheless remains very securely closed nearby).     

Axenic interspecies and intraclonal hybrid formation in Leishmania: Successful crossings between visceral and cutaneous strains

Diseases caused by trypanosomatids are serious public health concerns in low-income endemic countries. Leishmaniasis is presented in two main clinical forms, visceral leishmaniasis—caused by L. infantum and L. donovani—and cutaneous leishmaniasis—caused by many species, including L. major, L. tropica and L. braziliensis. As for certain other trypanosomatids, sexual reproduction has been confirmed in these parasites, and formation of hybrids can contribute to virulence, drug resistance or adaptation to the host immune system. In the present work, the capability of intraclonal and interspecies genetic exchange has been investigated using three parental strains: L. donovani, L. tropica and L. major, which have been engineered to express different fluorescent proteins and antibiotic resistance markers in order to facilitate the phenotypic selection of hybrid parasites after mating events. Stationary and exponential-phase promastigotes of each species were used, in in vitro experiments, some of them containing LULO cells (an embryonic cell line derived from Lutzomyia longipalpis). Several intraclonal hybrids were obtained with L. tropica as crossing progenitor, but not with L. donovani or L. major. In interspecies crossings, three L. donovani x L. major hybrids and two L. donovani x L. tropica hybrids were isolated, thereby demonstrating the feasibility to obtain in vitro hybrids of parental lines causing different tropism of leishmaniasis. Ploidy analysis revealed an increase in DNA content in all hybrids compared to the parental strains, and nuclear analysis showed that interspecies hybrids are complete hybrids, i.e. each of them showing at least one chromosomal set from each parental. Regarding kDNA inheritance, discrepancies were observed between maxi and minicircle heritage. Finally, phenotypic studies showed either intermediate phenotypes in terms of growth profiles, or a decreased in vitro infection capacity compared to the parental cells. To the best of our knowledge, this is the first time that in vitro interspecies outcrossing has been demonstrated between Leishmania species with different tropism, thus contributing to shed light on the mechanisms underlying sexual reproduction in these parasites.     

Two new rhabditida species (Nematoda: Rhabditidae) parasites of Cyclocephala signaticollis (Coleoptera: Scarabaeidae) in Argentina

Two different Rhabditida species (Nematoda: Rhabditidae) Parasitorhabditis platidontus n. sp. and Cruznema campestris n. sp. are described and illustrated from the larvae of Cyclocephala signaticollis (Coleoptera: Scarabaeidae) from Buenos Aires province, Argentina. Parasitorhabditis platidontus n. sp. is characterized by having 4 odontoplates in the stoma and the vulva is at 80% of the length of the body. Cruznema campestris n. sp. can be distinguished by the presence of 4 odontoplates and the arrangement of the male genital papillae, 9 pairs, of which 4 are preanal and 5 postanal.     

Sequence of the Small Subunit Ribosomal RNA Gene of Perkinsus atlanticus-like Isolated from Carpet Shell Clam in Galicia, Spain

Parasites identified as Perkinsus atlanticus have been reported infecting carpet shell clams in Galicia (northwest Spain). We have sequenced the 18S ribosomal RNA gene of in vitro cultured Perkinsus atlanticus-like or hypnospores from diseased clams, and compared it with the same genomic region from P. marinus and Perkinsus sp. We have also compared the sequence of internal transcribed spacer (ITS) 1, ITS 2, and 5.8S rRNA from our isolate with the P. atlanticus GenBank sequence. The phylogenetic analysis of our cultured parasite based on the 18S gene led us to conclude that this isolate is not related to the genus Perkinsus but to the protists Anurofeca, Ichthyophonus, and Psorospermium, located near the animal-fungal divergence. These last two genera have been included, together with Dermocystidium, in the newly described DRIPs (Dermocystidium, rossete agent, Ichthyophonus, and Psorospermium) clade, recently named Mesomycetozoa. Received October 25, 1999; accepted February 11, 2000.     

Deciphering the Acylation Pattern of Yersinia enterocolitica Lipid A

Pathogenic bacteria may modify their surface to evade the host innate immune response. Yersinia enterocolitica modulates its lipopolysaccharide (LPS) lipid A structure, and the key regulatory signal is temperature. At 21°C, lipid A is hexa-acylated and may be modified with aminoarabinose or palmitate. At 37°C, Y. enterocolitica expresses a tetra-acylated lipid A consistent with the 3′-O-deacylation of the molecule. In this work, by combining genetic and mass spectrometric analysis, we establish that Y. enterocolitica encodes a lipid A deacylase, LpxR, responsible for the lipid A structure observed at 37°C. Western blot analyses indicate that LpxR exhibits latency at 21°C, deacylation of lipid A is not observed despite the expression of LpxR in the membrane. Aminoarabinose-modified lipid A is involved in the latency. 3-D modelling, docking and site-directed mutagenesis experiments showed that LpxR D31 reduces the active site cavity volume so that aminoarabinose containing Kdo₂-lipid A cannot be accommodated and, therefore, not deacylated. Our data revealed that the expression of lpxR is negatively controlled by RovA and PhoPQ which are necessary for the lipid A modification with aminoarabinose. Next, we investigated the role of lipid A structural plasticity conferred by LpxR on the expression/function of Y. enterocolitica virulence factors. We present evidence that motility and invasion of eukaryotic cells were reduced in the lpxR mutant grown at 21°C. Mechanistically, our data revealed that the expressions of flhDC and rovA, regulators controlling the flagellar regulon and invasin respectively, were down-regulated in the mutant. In contrast, the levels of the virulence plasmid (pYV)-encoded virulence factors Yops and YadA were not affected in the lpxR mutant. Finally, we establish that the low inflammatory response associated to Y. enterocolitica infections is the sum of the anti-inflammatory action exerted by pYV-encoded YopP and the reduced activation of the LPS receptor by a LpxR-dependent deacylated LPS.     

Anaerobic co-digestion of swine manure with energy crop residues

Anaerobic co-digestion involves the treatment of different substrates with the aim of improving the production of biogas and the stability of the process. In this research, co-digestion of swine manure (SM) and energy crop residues (ECRs) was studied. The mixtures evaluated contained SM combined with maize (Mz), rapeseed (Rs) or sunflower (Sf) residues. Batch and semi-continuous experiments were performed to determine methane (CH₄) yields and the behavior of reactors while co-digesting agricultural wastes. Three different proportions of ECRs were tested in batch experiments for co-digestion with SM: 25, 50, and 75% volatile solids (VS). On the basis of the results obtained from batch tests, a mixture with a 50% ECR content was selected for the second stage of the study. Mesophilic reactors with a 3 L working volume were used for semi-continuous experiments. The hydraulic retention time (HRT) was set at 30 days and the reactors were kept under these operational conditions over four HRTs. The addition of ECR to the co-digestion system resulted in a major increase in the amount of biogas produced daily. The highest biogas yield was obtained when co-digesting Rs (3.5 L/day), although no improvement was observed in specific gas production from the addition of the co-substrate.     

Increasing synergistic effects of habitat destruction and hunting on mammals over three decades in the Gran Chaco

Habitat destruction and overexploitation are the main threats to biodiversity and where they co-occur, their combined impact is often larger than their individual one. Yet, detailed knowledge of the spatial footprints of these threats is lacking, including where they overlap and how they change over time. These knowledge gaps are real barriers for effective conservation planning. Here, we develop a novel approach to reconstruct the individual and combined footprints of both threats over time. We combine satellite-based land-cover change maps, habitat suitability models and hunting pressure models to demonstrate our approach for the community of larger mammals (48 species > 1 kg) across the 1.1 million km² Gran Chaco region, a global deforestation hotspot covering parts of Argentina, Bolivia and Paraguay. This provides three key insights. First, we find that the footprints of habitat destruction and hunting pressure expanded considerably between 1985 and 2015, across ~40% of the entire Chaco – twice the area affected by deforestation. Second, both threats increasingly acted together within the ranges of larger mammals in the Chaco (17% increase on average, ± 20% SD, cumulative increase of co-occurring threats across 465 000 km²), suggesting large synergistic effects. Conversely, core areas of high-quality habitats declined on average by 38%. Third, we identified remaining priority areas for conservation in the northern and central Chaco, many of which are outside the protected area network. We also identify hotspots of high threat impacts in central Paraguay and northern Argentina, providing a spatial template for threat-specific conservation action. Overall, our findings suggest increasing synergistic effects between habitat destruction and hunting pressure in the Chaco, a situation likely common in many tropical deforestation frontiers. Our work highlights how threats can be traced in space and time to understand their individual and combined impact, even in situations where data are sparse.     

Brucella β 1,2 Cyclic Glucan Is an Activator of Human and Mouse Dendritic Cells

Bacterial cyclic glucans are glucose polymers that concentrate within the periplasm of alpha-proteobacteria. These molecules are necessary to maintain the homeostasis of the cell envelope by contributing to the osmolarity of Gram negative bacteria. Here, we demonstrate that Brucella β 1,2 cyclic glucans are potent activators of human and mouse dendritic cells. Dendritic cells activation by Brucella β 1,2 cyclic glucans requires TLR4, MyD88 and TRIF, but not CD14. The Brucella cyclic glucans showed neither toxicity nor immunogenicity compared to LPS and triggered antigen-specific CD8⁺ T cell responses in vivo. These cyclic glucans also enhanced antigen-specific CD4⁺ and CD8⁺ T cell responses including cross-presentation by different human DC subsets. Brucella β 1,2 cyclic glucans increased the memory CD4⁺ T cell responses of blood mononuclear cells exposed to recombinant fusion proteins composed of anti-CD40 antibody and antigens from both hepatitis C virus and Mycobacterium tuberculosis. Thus cyclic glucans represent a new class of adjuvants, which might contribute to the development of effective antimicrobial therapies.     

Seminal plasma improves cryopreservation of Iberian red deer epididymal sperm

We tested the protective action of seminal plasma on epididymal spermatozoa from Iberian red deer, especially considering cryopreservation, as a means for germplasm banking improvement. We obtained seminal plasma by centrifuging electroejaculated semen, and part of it was thermically inactivated (denatured plasma; 55 degrees C 30 min). Epididymal samples (always at 5 degrees C) were obtained from genitalia harvested after regulated hunting, and pooled for each assay (five in total). We tested three seminal plasma treatments (mixing seminal plasma with samples 2:1): no plasma, untreated plasma and denatured plasma; and four incubation treatments: 32 degrees C 15 min, 5 degrees C 15 min, 5 degrees C 2h and 5 degrees C 6h. After each incubation, samples were diluted 1:1 with extender: Tes-Tris-Fructose, 10% egg yolk, 4% glycerol; equilibrated for 2h at 5 degrees C, extended down to 10(8) spz./mL and frozen. Sperm quality was evaluated before 1:1 dilution, before freezing and after thawing the samples, assessing motility (CASA) and viability (percentage of viable and acrosome-intact spermatozoa; PI/PNA-FITC and fluorescent microscopy). Plasma treatment, both untreated and denatured, rendered higher viability before freezing and higher results for most parameters after thawing. The improvement was irrespective of incubation treatment, except for viability, which rendered slightly different results for untreated and denatured plasma. This may be due to the presence of thermolabile components. We still have to determine the underlying mechanisms involved in this protection. These results might help to improve the design of cryopreservation extenders for red deer epididymal sperm.