Comparative transcriptome profiles of Schistosoma japonicum larval stages: Implications for parasite biology and host invasion

Schistosoma japonicum is prevalent in Asia with a wide mammalian host range, which leads to highly harmful zoonotic parasitic diseases. Most previous transcriptomic studies have been performed on this parasite, but mainly focus on stages inside the mammalian host. Moreover, few larval transcriptomic data are available in public databases. Here we mapped the detailed transcriptome profiles of four S. japonicum larval stages including eggs, miracidia, sporocysts and cercariae, providing a comprehensive development picture outside of the mammalian host. By analyzing the stage-specific/enriched genes, we identified functional genes associated with the biological characteristic at each stage: e.g. we observed enrichment of genes necessary for DNA replication only in sporocysts, while those involved in proteolysis were upregulated in sporocysts and/or cercariae. This data indicated that miracidia might use leishmanolysin and neprilysin to penetrate the snail, while elastase (SjCE2b) and leishmanolysin might contribute to the cercariae invasion. The expression profile of stem cell markers revealed potential germinal cell conversion during larval development. Additionally, our analysis indicated that tandem duplications had driven the expansion of the papain family in S. japonicum. Notably, all the duplicated cathepsin B-like proteases were highly expressed in cercariae. Utilizing our 3rd version of S. japonicum genome, we further characterized the alternative splicing profiles throughout these four stages. Taken together, the present study provides compressive gene expression profiles of S. japonicum larval stages and identifies a set of genes that might be involved in intermediate and definitive host invasion.

EXPANDING THE RECORD OF LARVAE OF FALSE FLOWER BEETLES WITH PROMINENT TERMINAL ENDS

Beetle larvae contribute to the overall biomass with a great share, yet they often stay unnoticed and underexplored. Larvae of the group Scraptiidae, also called false flower beetles, lead a life hidden in the wood, not easily accessible for observers. There, they contribute to wood decomposition and carbon cycling. Even though their ecological role is of great importance, these larvae have been comparably rarely studied. This is true for extant as well as fossil representatives of this group. It seems that this knowledge gap is not based on the limited availability of material but results from insufficiently studied material. Here we report new specimens, of which seven are extant and twelve are fossil. Fossil specimens are either from 40-million-year-old Baltic amber (Eocene) or 100-million-year-old Myanmar amber (Cretaceous), the latter representing the oldest record of these larvae. All specimens considered here possess a large, elongated terminal end. We performed an outline analysis of the shape of this terminal end for all so far known larval specimens sufficiently well preserved (in total 33 specimens: 17 extant, 14 Eocene, 2 Cretaceous). There is a recognisable difference between Eocene and extant specimens, yet it remains unclear whether this is due to different represented larval stages or an effect of evolution.

Comparative Studies of Renin-Null Zebrafish and Mice Provide New Functional Insights

Background: The renin-angiotensin system is highly conserved across vertebrates, including zebrafish, which possess orthologous genes coding for renin-angiotensin system proteins, and specialized mural cells of the kidney arterioles, capable of synthesising and secreting renin. Methods: We generated zebrafish with CRISPR-Cas9-targeted knockout of renin ( ren −/− ) to investigate renin function in a low blood pressure environment. We used single-cell (10×) RNA sequencing analysis to compare the transcriptome profiles of renin lineage cells from mesonephric kidneys of ren −/− with ren +/+ zebrafish and with the metanephric kidneys of Ren1 c−/− and Ren1 c +/+ mice. Results: The ren −/− larvae exhibited delays in larval growth, glomerular fusion and appearance of a swim bladder, but were viable and withstood low salinity during early larval stages. Optogenetic ablation of renin-expressing cells, located at the anterior mesenteric artery of 3-day-old larvae, caused a loss of tone, due to diminished contractility. The ren −/− mesonephric kidney exhibited vacuolated cells in the proximal tubule, which were also observed in Ren1 c−/− mouse kidney. Fluorescent reporters for renin and smooth muscle actin ( tg(ren:LifeAct-RFP; acta2:EGFP )), revealed a dramatic recruitment of renin lineage cells along the renal vasculature of adult ren −/− fish, suggesting a continued requirement for renin, in the absence of detectable angiotensin metabolites, as seen in the Ren1 YFP Ren1 c−/− mouse. Both phenotypes were rescued by alleles lacking the potential for glycosylation at exon 2, suggesting that glycosylation is not essential for normal physiological function. Conclusions: Phenotypic similarities and transcriptional variations between mouse and zebrafish renin knockouts suggests evolution of renin cell function with terrestrial survival.

Drosophila septin interacting protein 1 regulates neurogenesis in the early developing larval brain

Neurogenesis in the Drosophila central brain progresses dynamically in order to generate appropriate numbers of neurons during different stages of development. Thus, a central challenge in neurobiology is to reveal the molecular and genetic mechanisms of neurogenesis timing. Here, we found that neurogenesis is significantly impaired when a novel mutation, Nuwa, is induced at early but not late larval stages. Intriguingly, when the Nuwa mutation is induced in neuroblasts of olfactory projection neurons (PNs) at the embryonic stage, embryonic-born PNs are generated, but larval-born PNs of the same origin fail to be produced. Through molecular characterization and transgenic rescue experiments, we determined that Nuwa is a loss-of-function mutation in Drosophila septin interacting protein 1 (sip1). Furthermore, we found that SIP1 expression is enriched in neuroblasts, and RNAi knockdown of sip1 using a neuroblast driver results in formation of small and aberrant brains. Finally, full-length SIP1 protein and truncated SIP1 proteins lacking either the N- or C-terminus display different subcellular localization patterns, and only full-length SIP1 can rescue the Nuwa-associated neurogenesis defect. Taken together, these results suggest that SIP1 acts as a crucial factor for specific neurogenesis programs in the early developing larval brain.

Late effects of Beauveria bassiana on larval stages of Aedes aegypti Linneo, 1762 (Diptera: Culicidae)

Aedes aegypti is a culicide that has gained relevance over the years due to its ability to transmit various viruses that cause diseases in humans that all the years cause high mortality rates in the world population. The main problem is that Ae. aegypti has managed to establish and maintain a close relationship with humans and their habitat, which is why the search for alternatives to control vector populations becomes imperative. The objective of the present work was to study the effects of two Beauveria bassiana strains on Aedes aegypti. Third instar larvae of Ae. aegypti in 250 mL plastic containers were inoculated with the GHA and NB3 strains at different concentrations (1.5 × 104, 1.5× 105, 1.5 × 106 and 1.5 × 107 conidia/mL). The NB3 strain presented highest mortality values with 63% in the highest concentration i.e., 1.5 × 107, while for the GHA strain the highest mortality value was 30.7% at the same concentration. The results showed significant difference in mortality with respect to the strain and days post treatment (P = 0.0001), but not with respect to the conidial concentration (P = 0.634). The average mortality of larvae per day for the NB3 for different concentrations ranged from 20 to 25 larvae per day, while for the GHA daily mortality ranged from 5 to 12 larvae. In post-treatment mortality, the highest mortality was recorded in the third stage larvae for the NB3, while for GHA the highest percentage mortality was observed in individuals who managed to reach the adult state. The findings of the current research depicted the noteworthy role of B. bassiana for the management of an important vector of human disease.

Is Forensic Entomology Lost in Space?

Spatial and scale effects have barely been considered in forensic entomology, despite their pervasive influence on most of the parameters affecting the development of insect larval stages and the progression of insect succession on cadavers. Here, we used smoothing/interpolation techniques and semivariograms to document the spatial dynamics of sarcosaprophageous Calliphoridae, an important forensic taxon, in the Greater Moncton area in New Brunswick, Canada. Results indicated that the spatial dynamics of Calliphoridae differed between species, some species showing strong patterns of regional aggregation while others did not. Multivariate spatial correlations indicated that interspecific relationships in space varied widely, ranging from local and large-scale aggregation to spatial anticorrelation between species. Overall, this study suggested that even within a restricted timescale, the spatial dynamics of Calliphoridae can operate at many scales, manifest in different patterns, and be attributed to multiple different causes. We stress that forensic entomology has much to benefit from the use of spatial analysis because many important forensic questions, both at the fundamental and practical levels, require a spatial solution.

The origin and evolution of loqs2: a gene encoding an antiviral dsRNA binding protein in Aedes mosquitoes

Mosquito borne viruses such as dengue, Zika, yellow fever and Chikungunya cause millions of infections every year. These viruses are mostly transmitted by two urban-adapted mosquito species, Aedes aegypti and Aedes albopictus, that appear to be more permissive to arbovirus infections compared to closely related species. Although mechanistic understanding remains, Aedes mosquitoes may have evolved specialized antiviral mechanisms that potentially contribute to the low impact of viral infection. Recently, we reported the identification of an Aedes specific double-stranded RNA binding protein (dsRBP), named Loqs2, that is involved in the control of infection by dengue and Zika viruses in Ae. aegypti. Loqs2 interacts with two important co-factors of the RNA interference (RNAi) pathway, Loquacious (Loqs) and R2D2, and seems to be a strong regulator of the antiviral defense. However, the origin and evolution of loqs2 remains unclear. Here, we describe that loqs2 likely originated from two independent duplications of the first dsRNA binding domain (dsRBD) of loquacious that occurred before the radiation of the Aedes Stegomya subgenus. After its origin, our analyses suggest that loqs2 evolved by relaxed positive selection towards neofunctionalization. In fact, loqs2 is evolving at a faster pace compared to other RNAi components such as loquacious, r2d2 and Dicer-2 in Aedes mosquitoes. Unlike loquacious, transcriptomic analysis showed that loqs2 expression is tightly regulated, almost restricted to reproductive tissues in Ae. aegypti and Ae. albopictus. Transgenic mosquitoes engineered to ubiquitously express loqs2 show massive dysregulation of stress response genes and undergo developmental arrest at larval stages. Overall, our results uncover the possible origin and neofunctionalization of a novel antiviral gene, loqs2, in Aedes mosquitoes that ultimately may contribute to their effectiveness as vectors for arboviruses.

MOLECULAR CHARACTERIZATION OF ANISAKID NEMATODES HYSTEROTHYLACIUM SPECIES FROM JAPANESE THREADFIN BREAM NEMIPTERUS JAPONICUS (BLOCH, 1791) (PERCIFORMES, NEMIPERIDAE) FROM IRAQI MARINE WATER FISH

The present study provides a new insight into valuable information on the diverse structure of the Anisakid population and discusses the limited species richness in the Nemipterus japonicus (Bloch,1791) (Perciformes, Nemiperidae). The fishing area consists of various locations in the Arabian Gulf (29°58 0 33 00 N48°28 0 20 E). A total of 315 marine fish were examined, (n=287) were infected. Larval stages (n= 763) encysted within the mesenteries peritoneum and viscera of fish organs were isolated, with a prevalence of 91.11% of infection and, the intensity was 2.65. Molecular analysis was carried out on thirty individuals who have examined the morphology and showed some appearance differences, by amplifying internal transcribed spacers ITS and ITS-1 of nuclear rDNA (rDNA) by PCR using the primer sets NC5/NC2 and SS1/NC13R of thirteen DNA products. Evolutionary analyses were conducted in MEGA X. based on the identity percentage in the GenBank database showed that they belong to anisakid nematodes, in particular, they belong to eleven distinct taxa within the Hysterothylacium Ward & Magath, 1917 (Rhabditida, Raphidascarididae) and one identified species H. amoyense (Hsü, 1933) Deardorff & Overstreet, 1980. The current study records eleven species that belong to a genus of Hysterothylacium; some of the alignment of sequences polymorphisms reveals new different individuals of larvae species that may be adopted as new species if their adult stage is detected, and N. japonicus fish considered as a new host record. The current study provides some insights on the systematic taxonomy of these parasites, in addition, it supports similar studies that have been published elsewhere.

Response of Marine Plankton Communities in Ponds to the Presence of Vertical Structures

The effects of bottom vertical structures like AquaMats® in enhancing plankton productivity was evaluated. One experimental earthen pond of 500 m2 was provided with AquaMats® increasing the surface substrate area 12 times and water quality, phytoplankton and zooplankton populations developed during almost 100 days was compared with a pond without AquaMats®. Their presence favored the development of Dinoflagellates (Miozoa, Dinophyceae), mostly Gymnodiniales, which may be of some concern since some species of this group have been associated with toxic algal blooms while in the ponds without AquaMats® Diatoms (Bacillariophyta) predominate. In both ponds plankton production was very much sculptured by external nutrients added to the systems. The balance between different nutrients is extremely important to regulate the phytoplankton populations with Diatoms blooming at silicate concentrations higher than 2 μM and below this level and at low nitrate and high ammonium being more appropriate for Dinoflagellates. The linkage between phytoplankton and zooplankton population in ponds is strong with zooplankton exerting control over the phytoplankton population and vice-versa. The use of vertical substrates enhances plankton productivity by increasing the substrate area for periphyton fixation. The main zooplankton taxonomic groups associated with the presence of AquaMats® were Calanoid and Harpacticoid copepodites and nauplii, veligers of gastropods and trochophore of polychaets, larval stages of organisms that except for calanoid copepods are benthic and correspond to the meroplanktonic phase in the life cycle of those organisms.

Phylogenetic analysis and designing new primers for molecular identification of Drosophila suzukii

The family Drosophilidae includes over 3750 species worldwide and over 2000 of these are species of Drosophila. Spotted wing drosophila (SWD), Drosophila suzukii is one of the most dangerous species in this family. The insects live on undamaged ripening fruits, using its peculiar serrated ovipositor to break the skin of fresh ripening fruits and lay eggs in it. Drosophila species are very difficult and practically impossible to detect at larval stages. The present investigation was conducted at the All-Russian Plant Quarantine Center and Agrarian and Technological Institute of RUDN University, Moscow, Russia in 20182020. The aim of this study was to investigate the method of accurate and rapid identification of D. suzukii, and to design specific primer pairs for pest identification by Real-Time PCR method. The real-time quantitative PCR is a fast, sensitive, repeatable and accurate method for quantifying gene transcript levels. In this study, we designed specific primers (4.Dsuz.FRP) for Real-Time PCR to identify D. suzukii from other relative species. Although D. suzukii is absent in the Russian Federation and has not been reported so far, the project could be a precautionary measure.