Excretory Gland Research Articles

Serpentirhabdias mexicanus n. sp. (Nematoda: Rhabdiasidae), a parasitic lungworm of the nauyaca viper Bothrops asper (Serpentes: Viperidae) in the Mexican Neotropics

Serpentirhabdias mexicanusn. sp. (Nematoda: Rhabdiasidae) is described from the lung of the nauyaca viper Bothrops asper in Puebla State, central Mexico. This new species is the fifth of the genus described having onchia. Among the species included in this group, the new species is morphologically closest to S. viperidicus and S. atroxi. However, it differs from both species mainly by having only one excretory gland (compared to two present in S. viperidicus and S. atroxi). In addition, S. mexicanusn. sp. can be separated of S. viperidicus by tail length, shape of vulval lips, geographic distribution and host species and from S. atroxi by body length, number of papillae in the cephalic region, as well as the host species and geographic distribution. In the present study, we propose the new species based on morphological, host spectrum and genetic evidence. Phylogenetic analysis indicated Serpentirhabdias as a monophyletic group, with two subgroups that are congruent with the presence/absence of onchia in the esophagostome, host association and other relevant morphological characters.

Arterial and venous supply of the Harderian gland in hens.

The Harderian gland in domestic birds is a major paraocular excretory gland that has an important role in tear production as well as in the immune protection of the conjunctiva surface. The aim of this research was to investigate the arterial and venous supply of the gland in hens and provide valuable and useful information for future research. The research was conducted on 26 adult hens, provenience of Lohmann Brown. For the identification and determination of blood vessels, we used the vascular corrosion cast technique in conjunction with the transmission electron microscope (TEM). The casts showed that the gland receives the arterial supply via branches of a. ophthalmotemporalis and a. nasalis communis and these arteries are accompanied by the corresponding veins. Ultrastructural analyses showed the presence of fenestrated capillaries, which indicates the possibility for permeability of larger molecules. The present research gives important and detailed information about the arterial and venous supply of the Harderian gland in hens that may serve as guidelines for future vascular and morphological investigations.

Fecundity, in vitro early larval development and karyotype of the zoonotic nematode Anisakis pegreffii

The in vitro life cycle of zoonotic helminths is an essential tool for -omic translational studies focused on disease control and treatment. Anisakiosis is an emerging zoonosis contracted by the ingestion of raw or undercooked fish infected with the third stage larvae (L3) of two sibling species Anisakis simplex sensu stricto (s.s.) and Anisakis pegreffii, the latter being the predominant species in the Mediterranean basin. Recently, in vitro culture of A. pegreffii has been developed to enable fast and large-scale production of fertile adults. However, the conditions for larval development from hatching to infective L3 were not fulfilled to complete the cycle. Herein, we used a Drosophila medium supplemented with chicken serum and adjusted different osmolarities to maintain the culture of L3 hatched from eggs for up to 17 weeks. The highest survival rate was observed in the medium with the highest osmolarities, which also allowed the highest larval exsheathment rate. Key morphological features of embryogenesis and postembryogenesis studied by transmission electron microscopy revealed that the excretory gland cell is differentiated already up to 48 h post-hatching. Extracellular vesicles and cell-free mitochondria are discharged between the two cuticle sheets of the second stage larvae (L2). Contemporarly cultivated, two populations of adult A. simplex s.s. and A. pegreffii reached an average production of 29,914.05 (± 27,629.36) and 24,370.96 (± 12,564.86) eggs/day/female, respectively. The chromosome spreads of A. pegreffii obtained from mature gonads suggests a diploid karyotype formula of 2n = 18. The development of a reliable protocol for the in vitro culture of a polyxenous nematode such as Anisakis spp. will serve to screen for much needed novel drug targets, but also to study the intricated and unknown ecological and physiological traits of these trophically transmitted marine nematodes.

Effect of sublethal concentrations of glyphosate-based herbicides (Roundup Active®) on skin of the tropical frog (Dendropsophus molitor)

In Colombia, glyphosate (GP) is used to control weeds, with Roundup Active® being the most widely used. This use has affected aquatic ecosystems, causing malformations in amphibians. The Savannah frog (D. molitor) is a tropical frog inhabiting the mountain of Colombia. In the present study, we determined the effect of sublethal concentrations of GP (Roundup Active®) on the skin of D. molitor. Twenty-four tadpoles were exposed to concentrations of GP (T1: 0, T2: 1.4, T3: 3.6, and T4: 5.6 a.e mg/L) during 31 days. In 10 individuals per treatment, two skin regions were evaluated: dorsal cranial and caudal ventral to determine histopathological alterations. Morphometric analysis of the layers of the skin was performed: epidermis, dermis, and hypodermis-muscular. T1 did not present histopathological alterations. Since T2 was identified, glandular cell hyperplasia and hypertrophy increased melanophores and melanin accumulations in the highest concentrations of GP. The ultrastructure revealed an increase in excretory glands in the dermis. In the other layers, an increase of melanophores and melanocyte clusters was observed accompanied by vacuolization of basal cells. The morphometry showed an increase in the thickness of the dermis in the dorso-cranial region in T2 compared to the other treatments, while the ventral caudal region exhibited a variation in the thickness of the dermis from T2 and a decrease in T4. Despite evaluating sublethal concentrations, the skin of D. molitor tadpoles presents histopathological, ultrastructural, and morphometric alterations that could affect the survival of the species in the natural environment.

Chemical Composition and Biological Activity of Salvia officinalis L. Essential Oil.

In our study, we investigated the chemical composition and cytotoxic activity of essential oils isolated from Dalmatian sage (Salvia officinalis L.) collected along the Adriatic coast of Croatia. Scanning electron microscopy (SEM) was used to examine the morphology of the stem and leaf surfaces. Essential oil excretory glands were detected on both the leaves and stem surfaces. The essential oils were isolated by hydrodistillation, and their chemical composition was determined by gas chromatography and mass spectrometry (GC-MS). Sage essential oils were mixtures of terpene compounds, among which the most common were: α- and β-thujone, camphor, and 1,8-cineol. Cytotoxic activity was tested using MTS assay on multiple cell lines: normal and immortalized fibroblasts (HF77FA and HDF-Tert), immortalized lung line (BEAS-2B), and breast adenocarcinoma (MDA-MB-231). The growth of treated cells was determined relative to control conditions without treatment. The immortalized lung line was the least resistant to the activity of the essential oils, whereas immortalized fibroblasts were the most resistant. Statistical analysis has connected the cytotoxic effect and chemical composition of the studied essential oils. To the best of our knowledge, this work is the first testing of the cytotoxic activity of S. officinalis EO's on the BEAS-2B, HF77FA, and HDF-Tert cell lines. The presented data on essential oil chemical composition and cytotoxic effect on 4 types of human cells supports pharmacotherapeutic potential this plant is known to have.

Abstract 2098: Molecular based blood screening assay for metastatic adenoid cystic carcinoma

Abstract Adenoid cystic carcinoma (ACC) is a rare, yet highly lethal, tumor that originates in the excretory glands of the head and neck. It is a locally aggressive disease, with more than half of patients presenting with perineural invasion and/or bone erosion. The current standard of care consists of surgical excision followed by radiation. Many healthy patients are left with positive margins after surgery and receive systemic therapy in addition to adjuvant radiation without evidence showing added benefit for that approach. Despite extensive surgery, long-term survival rates remain low due to a high incidence of locoregional disease progression and metastatic disease to the lung, liver, brain, or bones, sometimes several years after definitive treatment. Extensive genome analysis of ACC has uncovered a high mutation rate of the proto-oncogene MYB, typically as a genomic translocation resulting in an MYB fusion event with one of the following gene loci: nuclear factor I/B (NFIB), transforming growth factor beta 3 (TGFBR3) or RAD51B. The translocation approximates an enhancer to the MYB regulatory element which can be transactivated by the MYB fusion product, thus leading to the overexpression of MYB via a positive feedback mechanism. Currently, detection of systemic disease relies on diagnostic imaging, which can be imprecise, costly and time-consuming. Years of routine imaging is very impractical, attrition rates are high, and patients often present years later with symptomatic metastatic disease. A rapid and sensitive molecular-based blood screening tool would allow for continuous monitoring and earlier detection of metastatic ACC, ideally improving survival outcomes for these patients. In this study, we hypothesized that the dysregulation in MYB, and the reliance of ACC cells on MYB for survival and progression could be harnessed to develop a molecular screening tool to monitor for metastatic disease of ACC. We developed a blood-based PCR screening probes for metastatic disease in patients with a history of ACC. By analyzing blood samples from four cohorts: healthy individuals (no history of cancer) (n=6), patients with a history of ACC but with no evidence of disease for longer than 3yrs (NED) (n=16), patients with newly diagnosed local disease (n=8), and patients with confirmed metastatic disease (n=7), we demonstrate that our assay was able to detect significantly elevated levels of aberrant MYB expression in all blood specimens from our metastatic disease cohort (p < 0.01). Additionally, our probes were able to detect aberrant levels of MYB in the blood of a patient 11 months before imaging techniques identified metastatic lung lesions. These findings indicate that we can harness a specific ACC genetic trait to develop a sensitive, rapid, and inexpensive molecular screening assay from blood to detect metastatic ACC. We continue to expand and validate these findings by screening existing and new ACC patients every 6 months. Citation Format: Acadia H. Moeyersoms, Ryan A. Gallo, Zoukaa B. Sargi, Jason M. Leibowitz, Andrew J. Rong, David T. Tse, Daniel Pelaez. Molecular based blood screening assay for metastatic adenoid cystic carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2098.

Three new species of Rhabdias Stiles et Hassall, 1905 (Nematoda: Rhabdiasidae) parasitic in Ptychadena spp. (Amphibia: Anura: Ptychadenidae) and an identification key to Rhabdias spp. from Afrotropical anurans.

Three new species of lung-dwelling nematodes are described from the frogs Ptychadena anchietae (Bocage), P. oxyrhynchus (Smith), and P. uzungwensis (Loveridge) in southern Africa. All three species are medium-sized species of Rhabdias Stiles et Hassall, 1905, with the thick-walled buccal capsules measuring 11-13μm×6-11μm, consisting of longer anterior and shorter posterior parts. Rhabdias athos n. sp. and R. porthos n. sp. are characterised by the rounded anterior end of the body and the presence of short dilatation of the oesophagus at its mid-length. Rhabdias porthos n. sp. has distinct excretory glands which are absent in two other species. Rhabdias aramis n. sp. is characterised by the truncated anterior end and the slight constriction of the oesophagus at the level of its mid-length. Phylogenetic analysis based on ITS-28S rDNA sequences placed R. aramis n. sp. in the clade consisting of R. engelbrechti Kuzmin et al., 2017 from South Africa and Eurasian Rhabdias spp., while R. athos n. sp. and R. porthos n. sp. formed a sister group to that clade. Identification key to 14 Rhabdias spp. parasitic in anuran amphibians from the Afrotropical Realm is provided.

PRIMARY SJÖGREN SYNDROME AND PULMONARY INVOLVEMENT: UTILITY OF SALIVARY GLAND BIOPSY IN DIAGNOSIS

PRIMARY SJÖGREN SYNDROME AND PULMONARY INVOLVEMENT: UTILITY OF SALIVARY GLAND BIOPSY IN DIAGNOSIS

A new species of Serpentirhabdias Tkach, Kuzmin et Snyder, 2014 (Nematoda: Rhabdiasidae) parasitic in the herald snake, Crotaphopeltis hotamboeia (Laurenti) (Reptilia: Serpentes: Colubridae) in South Africa.

Serpentirhabdias mamlambo n. sp. is described from the lung of the herald snake, Crotaphopeltis hotamboeia (Laurenti) in KwaZulu-Natal Province, South Africa. The new species is characterised by the small lips, the absence of the buccal capsule, the large excretory glands (1.5-2.3 times longer than the oesophagus), and the numerous eggs in uteri. Serpentirhabdias mamlambo n. sp. differs from other Serpentirhabdias spp. in the presence of M-shaped cuticular folds on dorsal and ventral sides of apical surface, the eye-shaped internal labial papillae, and the distinct dilatation of the oesophagus at its mid-length. Morphological differentiation of S. mamlambo n. sp. from its congeners is supported by molecular evidence based on cox1 mitochondrial gene, and ITS and partial 28S region of nuclear rDNA. Serpentirhabdias mamlambo n. sp. is the first species of the genus Serpentirhabdias described from the Afrotropical Realm.

The discovery of the lymphatic system in the 17th century. Part VII: the rise of vascular injection

In the second half of the seventeenth century, vascular injection was introduced in anatomy for the study of the mechanism of secretion of bodily fluids, a phenomenon into which the lymphatic system plays an important role. Injection became a routine procedure in the second half of the seventeenth century. Reinier de Graaf developed an appropriate syringe to inject liquid into minuscule tubules. He was the first to observe that water injected into seminiferous tubules was partially repelled by transudation to be absorbed by neighbouring lymph vessels. He also injected lymph vessels in and around the uterus and ovaries. His study friend Johannes Swammerdam developed a coloured hardening wax and Ruysch injected coloured hardening wax into vessels and ducts of lymph nodes and excretory glands. Ruysch introduced combined injection – corrosion procedures which resulted in delicate structures, including capillaries. He denied the presence of glandular structures in organs as described by Malpighi, and made blood vessels inclusive lymph vessels agents instead of aids to fluid secretion. His ideas resulted in the concept of the body being completely vascular, a theory which became commonplace in Dutch medical circles. Antony Nuck, the professor in medicine at the Leiden University, injected an amalgam of quicksilver and tin for further evaluation of the lymphatic system. He thought that lymph vessels originated from distal arteries and sustained that the shape of pores in these arteries determined the mechanisms of secretion in secretory glands and in lymph glands. He introduced lymphography.

Molecular cloning, expression and characterization of aspartyl protease inhibitor from Ancylostoma ceylanicum.

Aspartyl protease inhibitors (APIs) from parasitic intestinal nematodes are highly immunogenic and have been suggested as potential vaccine antigens. Ac-API-1 from Ancylostoma caninum showed strong immunogenicity and its polyclonal antibodies could specifically recognize the excretory/secretory products of adult worms. However, little is known about molecular characteristics and biological function of API from Ancylostoma ceylanicum (Ace-API). In this study, the Ace-API mature peptide coding sequence was cloned and expressed, and molecular characteristics of its full length sequence were analyzed. Ace-API cDNA was 684bp in length, which encoded 228 amino acids. The similarity of the Ace-API amino acid sequence to Ac-API-1 and Adu-API-1 was 96.93% and 96.49%, respectively, and they clustered together in the phylogenetic tree. Escheria coli-expressed recombinant protein was mainly soluble in the supernatant of bacterial cell lysate. Western blot showed that Ace-API protein had good reactivity to the serum of infected dogs. Pepsin inhibition assay revealed that the recombinant protein had inhibitory activity on pepsin. Immunofluorescence results demonstrated that Ace-API was mainly localized to the epidermis, excretory glands, and pseudocoelomic fluid of the adult. Using the quantitative real-time PCR, the expression of Ace-api mRNA in adults was significantly higher than that in the third stage (L3) larvae. Together, these data indicate that Ace-API is secreted extracellularly by the parasite, and might play a role in protecting the parasite against the proteolytic digestion by the host proteases, which stimulate further studies to explore this protein as a potential hookworm vaccine candidate.

OP0302 IANALUMAB (VAY736), A DUAL MODE OF ACTION BIOLOGIC COMBINING BAFF RECEPTOR INHIBITION WITH B CELL DEPLETION, REACHES PRIMARY ENDPOINT FOR TREATMENT OF PRIMARY SJOGREN’S SYNDROME

Background:Primary Sjogren’s syndrome (pSS) is a multi-organ autoimmune disease mainly affecting excretory glands and characterised by B-cell hyperactivity. No approved systemic treatment is available. Ianalumab (VAY736) is an anti-B-cell activating factor (BAFF) receptor fully human monoclonal antibody, engineered for direct ADCC-mediated B-cell depletion.Objectives:This phase 2b study aimed at establishing a dose-response relationship over a range of VAY736 doses, using change from baseline (BL) in EULAR Sjogren’s Syndrome Disease Activity index (ESSDAI) over 24 Weeks (Wks) as primary endpoint. The study is ongoing with a second blinded treatment period up to Wk52. Here we report efficacy and safety Wk24.Methods:190 patients (pts) were randomised 1:1:1:1 to receive monthly s.c. doses of VAY736 (5, 50, 300mg) or placebo (PBO). Prior to 1st-dose of study treatment, pts received methylprednisolone i.v. 250mg. Eligible pts fulfilled American European Consensus Group (AECG) criteria, were anti-Ro/SSA+, had ESSDAI ≥6 and EULAR Sjogren’s Syndrome Patient Reported Index (ESSPRI) ≥5. Statistical methods included MCP-Mod to assess dose-response on change of ESSDAI from BL and responder rate analysis to calculate the proportion of pts with ≥3 points improvement on ESSDAI. Secondary endpoints included ESSPRI, Functional Assessment of Chronic Illness Therapy Fatigue (FACIT-F), Physician’s (PhGA) and Patient’s Global Assessments (PaGA), SF-36, stimulated salivary flow (sSF), Schirmer’s test.Results:Primary endpoint was met with statistically significant dose-response for ESSDAI (Figure). The largest ESSDAI reduction was 1.92 points over PBO for VAY736 300mg at Wk24. Responder rate analysis on ESSDAI revealed for 300mg vs PBO responder rates of 42/47 (89.4%) vs 30/49 (61.2%), a difference of 28.1% (p=0.0019). No differences were seen for 5mg and 50mg vs PBO. PhGA change from BL was significantly different between 300mg and PBO (p=0.022). A numerical trend for sSF improvement for VAY736 300mg compared to PBO was notable at Wk24 (p=0.092). For secondary endpoints ESSPRI and FACIT-F, VAY736 treatment showed no benefits over PBO. PBO responses were generally high. Incidence of treatment emergent AEs was comparable across all studied groups, whereby site injection reactions were most frequent, mostly mild and showed a dose-response.Conclusion:Primary endpoint assessing ESSDAI was met, showing statistically significant dose-response for ianalumab with clinically important improvement for 300mg vs PBO. Preliminary safety profile of ianalumab was good.Figure.ESSDAI Change from Baseline over Time up to Week 24 Reveals a Statistically Significant Dose Response RelationshipDisclosure of Interests:Thomas Dörner Grant/research support from: Janssen, Novartis, Roche, UCB, Consultant of: Abbvie, Celgene, Eli Lilly, Roche, Janssen, EMD, Speakers bureau: Eli Lilly, Roche, Samsung, Janssen, Simon J. Bowman Consultant of: Astrazeneca, Biogen, BMS, Celgene, Medimmune, MTPharma, Novartis, Ono, UCB, xtlbio, Glapagos, Speakers bureau: Novartis, Robert Fox Consultant of: Novartis, Pfizer and Lilly, Xavier Mariette Consultant of: BMS, Gilead, Medimmune, Novartis, Pfizer, Servier, UCB, Athena Papas Grant/research support from: Novartis, Consultant of: Novartis, Thomas Grader-Beck Grant/research support from: Abbvie, Celgene, Consultant of: Novartis, Lilly, Ben A Fisher Consultant of: Novartis, Roche, BMS and Servier, Filipe Barcelos Consultant of: Pfizer and Lilly, Salvatore De Vita Consultant of: Roche, Human Genome Science, Glaxo Smith Kline and Novartis, Hendrik Schulze-Koops Grant/research support from: Pfizer Inc, Robert J Moots: None declared, Guido Junge Shareholder of: Novartis, Employee of: Novartis, Janice Woznicki Shareholder of: Novartis, Employee of: Novartis, Monika Sopala Shareholder of: Novartis, Employee of: Novartis, Wen-Lin Luo Shareholder of: Novartis, Employee of: Novartis, Wolfgang Hueber Shareholder of: Novartis, Employee of: Novartis

Nosocomial Infections in Gastrointestinal Cancer Patients: Bacterial Profile, Antibiotic Resistance Pattern, and Prognostic Factors.

BackgroundCancers of the gastrointestinal (GI) tract and its associated excretory glands are one of the most common causes of cancer-related death worldwide, and these patients are more likely to developing nosocomial infections due to immunodeficiency.ObjectiveTo explore the bacterial profile, antibiotic resistance pattern, and prognostic factors of nosocomial infections in hospitalized GI cancer patients.MethodsAll electronic medical records of nosocomial infection episodes in hospitalized GI cancer patients were retrospectively reviewed. In-hospital mortality was used to evaluate the prognosis of patients. Mann–Whitney test, Chi-square test, and binary logistic regression analysis were used to identify potential risk factors for in-hospital mortality. P-values <0.05 were considered statistically significant.ResultsA total of 428 GI cancer patients developed nosocomial infections during hospitalization. Respiratory tract infections (44.2%), bloodstream infections (BSIs) (11.7%), and abdominal cavity infections (11.4%) were the most common infection sites. The predominant causative pathogens were extended-spectrum β-lactamase (ESBL)-producing Escherichia coli (13.6%), ESBL-negative E. coli (11.9%), and Klebsiella pneumoniae (10.0%). Multidrug-resistant (MDR) strains were detected in 27.6% of isolates. Antimicrobial susceptibility analysis showed that the isolated Gram-negative bacteria (GNB) exhibited high sensitivity to amikacin, meropenem, imipenem, and piperacillin/tazobactam, while the isolated Gram-positive bacteria exhibited high sensitivity to tigecycline, linezolid, and vancomycin. The overall in-hospital mortality of all patients was 11.2% in the study. Multivariate analysis showed that ECOG performance status ≥two scores, length of antibiotic treatment <9.0 days, existence of septic shock, and hypoproteinemia were independent risk factors for in-hospital mortality.ConclusionThe burden of nosocomial infections in GI cancer patients is considerably high, with GNB being predominantly isolated causative pathogens. Surveillance on serum albumin level, adequate antibiotic treatment, early identification, and prompt treatment of septic shock could benefit the prognosis.

The anatomy of the third-stage larva of Toxocara canis and Toxocara cati.

This review describes the morphology and ultrastructure of the third-stage larva of Toxocara canis, the larval stage that hatches from the egg and persists in the tissues of various paratenic hosts including infected humans. This larva remains unchanged as it passes from host to host and lives for extended periods in this same body form until it makes its way to the gastro-intestinal tract of its final host. There has been less work on the anatomy and ultrastructure of the larva of Toxocara cati, but overall, it appears very similar to that of T. canis with the only apparent difference being that the larva is a few microns thinner in diameter. The paper describes the anatomy and ultrastructure of the larva and the various major structures of the various body systems including the cuticle, hypodermis, the nervous tissue, oesophagus and the lumen-free intestine, and the large excretory gland cell that fills much of the pseudocoel and which appears to be the source of the components that are recognized immunologically by the infected host. It is suggested that further detailed studies on the larva could provide significant information that would allow a greater understanding of how the larva persists within these hosts and may provide information that would make the larva an excellent link to work on nematode anatomy that would link the work being done with this model system to that of Caenorhabditis elegans and the large amount of historical work already performed on adult ascaridoid nematodes.

Functional Ultrastructure of the Excretory Gland Cell in Zoonotic Anisakids (Anisakidae, Nematoda).

Excretory and secretory products are crucial for parasite infectivity and host immunomodulation, but the functioning and ultrastructure of the excretory gland cell (EC) that produces these products are still scarcely understood and described. In light of growing reports on anisakiasis cases in Europe, we aimed to characterise the EC of larval Anisakis pegreffii and adult Pseudoterranova azarasi. In the latter, EC starts 0.85 mm from the head tip, measuring 1.936 × 0.564 mm. Larval EC shows a long nucleus with thorn-like extravaginations toward the cytoplasm, numerous electron-dense and -lucent secretory granules spanning from the perinuclear to subplasmalemmal space, an elevated number of free ribosomes, small, spherical mitochondria with few cristae and a laminated matrix, small and few Golgi apparatuses, and few endoplasmic reticula, with wide cisternae complexes. Ultrastructure suggests that anaerobic glycolysis is the main metabolic pathway, obtained through nutrient endocytosis across the pseudocoelomic surface of the EC plasmalemma and its endocytic canaliculi. Thorn-like extravaginations of EC karyotheca likely mediate specific processes (Ca2+ signaling, gene expression, transport, nuclear lipid metabolism) into the extremely wide EC cytosol, enabling focal delivery of a signal to specific sites in a short time. These functional annotations of parasitic EC should help to clarify anisakiasis pathogenesis.

Description, molecular characterization and life cycle of Serpentirhabdias mussuranae n. sp. (Nematoda: Rhabdiasidae) from Clelia clelia (Reptilia: Colubroidea) in Brazil.

Serpentirhabdias mussuranae n. sp. is described from the lungs of the mussurana, Clelia clelia (Daudin, 1803), from vicinities of Lábrea, Amazonas State, Brazil. The species is characterized by the triangular oral opening, the presence of teeth (onchia) in the oesophastome, the excretory glands longer than the oesophagus and the tail abruptly narrowing in its anterior half and gradually tapering in posterior half. Among the Neotropical representatives of the genus, three species are known to possess the onchia in the oesophastome: S. atroxi, S. moi and S. viperidicus. Serpentirhabdias mussuranae n. sp. differs from S. atroxi and S. viperidicus by its triangular shape of the oral opening and the oesophastome in apical view, vs. round in the latter two congeners. Additionally, S. viperidicus has a larger oesophastome, 13-22 micrometers wide and 13-23 micrometers deep. The new species has relatively longer excretory glands than S. moi. The new species is morphologically and genetically close to S. atroxi, S. moi and S. viperidicus, all parasitic in Brazilian snakes, based on the presence of onchia and the comparison of nucleotide sequences of nuclear ribosomal DNA and mitochondrial cox1 gene (differences varied between 3.8% and 7.1%). Data on the life cycle of S. mussuranae n. sp. is provided, and the life cycle is typical of the genus Serpentirhabdias, with the combination of direct development and heterogony. Free-living larval stages and the adults of amphimictic free-living generation are described. The results of molecular phylogenetic analysis based on nuclear ribosomal internal transcribed spacer (ITS) + partial 28S region and partial mitochondrial cox1 gene are provided.

The discovery of the lymphatic system in the seventeenth century. Part V: an ode to the nerves

The English anatomists Francis Glisson and Thomas Wharton introduced theories on the use of recently discovered lymphatic system in the1650s. Their main idea was that membranous tissues were supplied by nerves with a vital fluid produced within nutritive glands, to be carried to the brain and thence from the brain to all membranous tissues, including glands. They stated that the distribution of the vital fluid was based on the similarity between its ingredients and the needs of the recipient tissues, and not on the weight, size or location of pores encountered, as maintained by Descartes. Lymph, a mixture of waste from the consumed vital fluid and moisture transuded by arterial capillaries, was absorbed by the lymphatic vessels to be excreted via excretory glands, or to be diverted to the venous system by reductive glands. The theories of Glisson and Wharton were very soon rejected to be replaced by a mechanistic philosophy, a legacy of Descartes’ theories.

Description, biology and molecular characterisation of Serpentirhabdias moi n. sp. (Nematoda: Rhabdiasidae) from Chironius exoletus (Serpentes: Colubridae) in Brazil

A new species of the genus Serpentirhabdias Tkach, Kuzmin et Snyder, 2014, S. moi n. sp., is described from a colubroid snake Chironius exoletus from Caxiuanã National Forest, State of Pará, Brazil. The species is characterised by having a triangular oral opening, absence of the buccal capsule, presence of six minute onchia in the oesophastome, and excretory glands of approximately the same length as the oesophagus. These qualitative morphological characters, as well as some measurements, differentiate the new species from other Neotropical and Nearctic Serpentirhabdias spp. The morphological description of parasitic adults of S. moi n. sp. is complemented by the description of free-living stages including males, females, and infective larvae. Comparative analysis of partial sequences of cox1 and 12S mitochondrial genes strongly supported the status of S. moi n. sp. as a new species. Molecular phylogeny based on sequences of the nuclear DNA region spanning the 3′ end of the 18S nuclear rRNA gene, ITS region (ITS1 + 5.8S + ITS2) and 5′ end of the 28S gene supported monophyly of all rhabdiasid genera included in the analysis and placed the new species into the Serpentirhabdias clade as sister taxon to S. fuscovenosa.

The morphology of free-living stages and immature parasites of Rhabdias paraensis (Nematoda: Rhabdiasidae), a parasite of Rhinella marina (Anura: Bufonidae) in Brazil.

Rhabdias paraensis Santos, Melo, Nascimento, Nascimento, Giese et Furtado, 2011 was described based on fully gravid worms. Further investigations on the free-living stages, immature worms and young individuals were facilitated by cultivation in the laboratory, which allowed us to add new information about the morphology and development of the species. Observations on the free-living development of R. paraensis showed that the life cycle is typical of Rhabdias, with alternation of gonochoristic and hermaphroditic generations and without homogony. Males of the free-living generation were different from those in several species of the genus studied previously. In the original description, the excretory glands and duct were absent in gravid specimens of R. paraensis, while in this study, distinct excretory glands and a duct were observed in immature and young individuals. Additionally, we recognised the separation of the buccal capsule walls into anterior and posterior portions and described the specific shapes of these portions in lateral and apical view. Studies on the morphology and development of free-living stages of Rhabdias spp. from Neotropical regions may provide additional information for species determination.

Phytoextraction of Cd by Atriplex nummularia L., a xero-halophyte species

This work is in the perspective to test if the species halo-xerophyte Atriplex nummularia L. which is a perennial shrub type C4 metabolism present in addition to its water resistance capacity and salt stress, a resistance metal stress. Atriplex nummularia plants were exposed to a mixture of cadmium (20 microM) and a range of increasing concentrations of NaCl [0; 34.2 mm (2 g / l) and 102.6 mm (6 g / l)]. After the analysis of physiological parameters such as dry matter and the amount of total chlorophylls, it has interested to become of the metal in the plant. Again, it has determined the amount of cadmium accumulated in the different organs of the plant and the amount excreted by the leaves. The results obtained show that Atriplex nummularia plant salt tolerance can also accumulated the cadmium. While the cadmium content of roots does not vary regardless of amount of NaCl present in the medium, these of leaves increases with the addition of salt and proportionally to the amount of NaCl in the medium. Cadmium excretion by excretory glands present in these leaves was observed and quantified. This characterization of the properties of completely original Atriplex nummularia vis-a-vis cadmium allows to be considered this xero-halophytic plant as a new potential candidate for the use in phytoremediation strategies of soil contaminated with heavy metals and more precisely phytoextraction.