Adhesive Disc Research Articles

A new species of Trichodina Ehrenberg, 1830 (Ciliophora: Trichodinidae) from Rhamdia quelen (Siluriformes: Heptapteridae) and Gymnotus sp. (Teleostei: Gymnotidae) in Brazil.

The present study describes a new species of Trichodina found in the mucus of the body surface of 137 farmed and wild silver catfish (Rhamdia quelen) located in southern Brazil and in the gills of 99 banded knife fish (Gymnotus sp.) captured in the basin of Pantanal. Smears of the body surface mucus and gills were air dried at room temperature and impregnated with silver nitrate and stained with Giemsa for analysis of the adhesive disc, denticles and macronucleus. Characterized as a medium trichodinid, Trichodina quelenii n. sp. resembles T. heterodentata and T. acuta. It differs from T. heterodentata by the shape of the blade and from T. acuta by the lack of central circle. Furthermore, the distinct shape of the blade of T. quelenii n. sp. differs from all previously described species for the genus, leaving no doubt it is a new species.

Modified Corneosurfametry as a new accelerated high-throughput ex vivo methodology for predicting cleanser effects towards human skin.

Corneosurfametry (CSM) was originally developed as a tool to predict irritation potential of cleansers. In this method, surface skin stripped using cyanoacrylate is contacted with surfactants/products, rinsed and stained with toluidine blue and basic fuschin dyes. The intensity of staining increases with increases in irritation potential of surfactant. Our objective was to modify the CSM technique to achieve better control of the tape stripping process. Another objective was to correlate the modified CSM (MCSM) with a traditional in-vivo forearm controlled application test (FCAT) for mildness and to explore its utility to assess the state of corneum after a clinical test. Surface skin cells were tape stripped from forearms of volunteers with D-Squame Adhesive Discs. Discs were treated with a 10% solution of the product in a 96-plate well for 10 min, rinsed, dried and treated with basic fuschin-toluidine blue dye solution, rinsed and dried again. Forearm Controlled Application Test (FCAT) was based on a published protocol. Tape strips obtained after product treatment were also analyzed by the MCSM procedure without additional product treatment. Mildness/barrier damage assessed from in-vivo FCAT showed a similar ranking to the MCSM results. MCSM, TEWL and Erythema analysis of between-treatment differences showed a good correlation indicating that barrier damage seen in in-vivo studies can be predicted from ex-vivo MCSM studies. MCSM analysis of tape strips after the FCAT study showed that the damage decreased with increase in tape strip number. A moisturizing body wash (MBW) with mild surfactants showed the least damage in all layers. In contrast, harsh dish washing liquid showed significantly higher damage down to several layers. Another MBW with petrolatum in a harsher base showed damage almost similar to that of the harsh dish washing liquid in the surface layers. Thus, the MCSM was able to show underlying damage which would have been normally masked by the deposited petrolatum. The MCSM assay was shown to be a valuable tool for accelerated high throughput evaluation of mildness of surfactants and fully formulated products. MCSM can also be used to assess the state of the corneum after a product treatment.

Differential localization and functional specialization of centrin analogs in the parasitic ciliate Trichodina pediculus.

Trichodinids are ciliated protozoans that reversibly attach to the tegument of marine and freshwater host-organisms via an adhesive disc. In this study, we have used permeabilized cell models of Trichodina pediculus to examine the distribution of centrins, a Ca(2+)-binding protein associated with centrioles and/or contractile filamentous structures in a large number of protists. The previous finding that filamentous material of the adhesive disc comprised a 23-kDa centrin analog suggested that this protein might be a disc-specific isoform. This possibility was explored through immunolabeling methods using two distinct antibodies, anti-ecto-endoplasmic boundary (EEB) and anti-Hscen2 previously shown to react respectively with centrin-based filament networks and with centrioles. Immunofluorescence microscopy showed that anti-EEB reacts with filamentous material of the disc but not with basal bodies. Conversely, anti-Hscen2 cross-reacted with basal bodies but failed to label any type of structure occurring in the disc area. More detailed data on localization of this protein was obtained by immunoelectron microscopy showing gold particles deposits in the lumen of basal bodies. The different patterns revealed by this immunochemical approach suggest that the two protein antigens concerned by this study are distinct centrin isoforms that presumably perform organelle-specific function in the ciliate T. pediculus.

Encystation of Giardia intestinalis—a Journey from the Duodenum to the Colon

The intestinal protozoan parasite Giardia intestinalis has a simple life cycle consisting of disease-causing trophozoites and infectious cysts. The parasite differentiates into the cyst stage (encystation) when duodenal trophozoites are swept further down in the small intestine. The encystation process can be divided into an early and a late phase. Expressions of cyst wall sugars and proteins are induced early during encystation, and Giardia has developed special transport systems for cyst wall material. This is combined with disassembly of the flagella and adhesive disc. In the later phase of encystation, nuclei are divided, DNA is replicated, and the parasites prepare for the next step in differentiation (excystation) via large changes in gene expression. Even if many aspects of the encystation process have been studied and much information has been gained during the last years, much still remains to be explored regarding this highly complex process.

Trichodina modesta: an exotic ciliate in the Neotropical region parasitizing an unusual host.

In this study, an important ornamental fish, Betta splendens (Osphronemidae), from three different Brazilian states was examined for parasitic infestations. Smears with parasites were impregnated with silver nitrate or stained using Giemsa for taxonomic evaluation. A disc-shaped trichodinid with a body diameter of 39.7 ± 3.3 µm, adhesive disc diameter of 32.9 ± 3.1 µm and denticulate ring diameter of 19.5 ± 2.0 µm was found. The morphological characteristics resembled those of Trichodina modesta Lom, 1970, a species that shows clear host specificity for Cypriniformes. Until now, its occurrence was restricted to the Eurasian region. In the present study, a new host for T. modesta is reported and therefore the first occurrence of this species in the Americas. The parasite was possibly introduced into the Neotropical region through the exotic fish trade, especially of Cypriniformes used by aquarists. The distribution of this ciliate is discussed and a checklist of localities and hosts for the species is provided.

Helium ion microscopy and ultra-high-resolution scanning electron microscopy analysis of membrane-extracted cells reveals novel characteristics of the cytoskeleton of Giardia intestinalis.

Giardia intestinalis presents a complex microtubular cytoskeleton formed by specialized structures, such as the adhesive disk, four pairs of flagella, the funis and the median body. The ultrastructural organization of the Giardia cytoskeleton has been analyzed using different microscopic techniques, including high-resolution scanning electron microscopy. Recent advances in scanning microscopy technology have opened a new venue for the characterization of cellular structures and include scanning probe microscopy techniques such as ultra-high-resolution scanning electron microscopy (UHRSEM) and helium ion microscopy (HIM). Here, we studied the organization of the cytoskeleton of G. intestinalis trophozoites using UHRSEM and HIM in membrane-extracted cells. The results revealed a number of new cytoskeletal elements associated with the lateral crest and the dorsal surface of the parasite. The fine structure of the banded collar was also observed. The marginal plates were seen linked to a network of filaments, which were continuous with filaments parallel to the main cell axis. Cytoplasmic filaments that supported the internal structures were seen by the first time. Using anti-actin antibody, we observed a labeling in these filamentous structures. Taken together, these data revealed new surface characteristics of the cytoskeleton of G. intestinalis and may contribute to an improved understanding of the structural organization of trophozoites.

Functional Morphology of the Remora Adhesive Disc

Remoras (Family Echeneidae) are best known for their cranial suction disc modified from dorsal fin elements. Remoras use the disc to adhere to a number of different hosts (e.g. sharks, fish, turtles, marine mammals, and ship hulls) for several possible reasons, including efficient travel, feeding opportunities, and locating mates. While previous work has shown that remoras can generate suction forces opposing pull-off removal, our research has found that the remora disc is a complex hierarchical mechanism with many different factors contributing to the long term but easily reversible nature of the remora adhesive disc. Here we focus on two morphological features that are responsible for maintaining attachment to a host: lamellar spinules and cranial vasculature. Using a friction model to estimate the spinule contribution to shear resistance, we found that the tooth-like spinules on the flat lamellae that make up the disc generate friction an order of magnitude greater when interacting with a rough substrate like shark skin than with a smooth surface, thereby explaining the difference in adhesive strength on different surfaces observed in previous work. The cranial vasculature of remoras is highly modified with respect to the position and relative size of anterior veins as compared to other fishes. Most notably, the anterior cardinal sinus lies dorsal to the cranium, in direct contact with the ventral surface of the disc. Presumably the orientation of the remora cranial vessels contributes to applying pressure on the disc against the host to maintain strong adhesion for extended periods of time.

Ontogenetic metamorphosis and extreme sexual dimorphism in lumpsuckers: Eumicrotremus asperrimus, Cyclopteropsis bergi and Cyclopteropsis lindbergi, may be synonymous.

Cyclopterids were hatched from egg batches that were laid in two empty buccinid snail shells collected from a depth of 340 m in the Sea of Japan. Larvae were reared to identify species and to describe the morphological changes associated with metamorphosis. The fin rays of all fins were mostly complete and the pelvic fins were modified to form suckers, or adhesive discs, at the time of hatching. Juveniles immediately attached themselves to the bottom and there was no planktonic stage. The body surface was smooth with no spines or bony tubercles. At 4 months after hatching, the fine spines present on the head and trunk of juveniles transformed into bony tubercles. At 7 months after hatching, fishes became sexually dimorphic including the position and development patterns of bony tubercles. Importantly, these sexually dimorphic changes in morphology corresponded closely with descriptions of different species. Specifically, females could be classified as Eumicrotremus asperrimus, and young and fully developed males as Cyclopteropsis bergi and Cyclopteropsis lindbergi, respectively. These observations resolved a previously ambiguous hypothesis regarding the taxonomy of these cyclopterid taxa. Cyclopteropsis bergi and C. lindbergi may be synonyms of E. asperrimus.

Trichodinid Parasites on the Gills ofChanna punctatusfrom the Wild and Cultured Environments in Sylhet, Bangladesh

Two trichodinid parasites,Trichodina cyprinocolaandTrichodina pediculus, were acknowledged in the gills during a sampling preparation of thirty fishChanna punctatusfrom the wild and cultured environments. However,Trichodina pediculuswas recorded only in one farmed fish. From a total of 33 parasites, 26.67% were encountered in wild and 46.67% in cultured fish. The overall incidence of infection byTrichodina cyprinocolawas 33.33% whilstTrichodina pediculuswas only 3.33%. Records from statistical analysis as well affirmed that fish host from the cultured environment constituted the highest rate of intensity(3.43±1.4), density(1.6±2.0), and infection index (11.2±13.95)values. Both trichodinid species are identified based on the study in their unique denticles structure. Relatively large size of adhesive disc with tiny curved denticles and elongated denticle rays are the exceptional denticle morphology observed inTrichodina pediculusand compact association of sickle denticles with round tangent point alongside short vigorous upright rays evaluated theTrichodina cyprinocola.

Structural Changes of Adhesive Discs during Attachment of Boston Ivy

This study investigates the developmental pattern of adhesive discs (ADs) to highlight the ontogeny and structural changes that occur during the growth of Boston ivy. Initiation to postmortem features of ADs were examined through light and scanning electron microscopy. The study also reveals a new finding of the dislocation of peripheral tissues of adaxial origin. Four phases of attachment are suggested with regards to its climbing behavior: 1) pre-attachment, 2) upon attachment, 3) after attachment, and 4) final attachment. During initiation, several ADs originate from tendril primordia without epidermal differentiation. However, different growth rates in the epidermis results in completely different ADs. ADs were discerned by size, shape, and color during expansion, but cells in the adaxial surface remained alive longer than the other side. Upon contact, the ADs demonstrate simultaneous growth and deterioration, but once attachment is established the latter process subdues to final stages. Epidermal transformation, adhesive secretion, cellular disruption, and mechanical stress were essential for the self-clinging nature of Boston ivy. The post-attachment sequence is also believed to be critical in achieving maximum mechanical strength to provide extensive support. The developmental process of ADs is prompted by tactile stimulation but in a highly organized and systematic manner.

Therapeutic enhancement of newly derived bacteriocins against Giardia lamblia

Trials for identifying efficient anti-giardial agents are still ongoing. Nowadays, bacteriocins have attracted the attention as potential antimicrobial compounds. For the first time, the current study evaluated the therapeutic efficacy of bacteriocins derived from newly isolated Egyptian strains of probiotics Lactobacilli; L. acidophilus (P106) and L. plantarum (P164) against Giardia lamblia. Bacteriocins' efficacy was evaluated both in vitro; by growth inhibition and adherence assays, and in vivo; through estimation of parasite density, intestinal histopathological examination and ultrastructural analysis of Giardia trophozoites. In vivo bacteriocins' clinical safety was assessed. In vitro results proved that 50 µg of L. acidophilus bacteriocin induced reduction of the mean Giardia lamblia trophozoites by 58.3 ± 4.04%, while at lower concentrations of 10 and 20 µg of both L. acidophilus and L. plantarum, non significant reduction of the mean parasite density was achieved. In vitro trophozoites adherence was susceptible to the tested bacteriocins at all studied concentrations with variable degrees, while the highest adherence reduction was demonstrated using 50 µg of L acidophilus bacteriocin. In vivo, oral inoculation of 50 µg/mouse L. acidophilus bacteriocin for 5 successive days resulted in a noteworthy decline of the intestinal parasite density, along with amelioration of intestinal pathology of infected mice. Ultrastructural examination proved thatfive doses of L. acidophilus bacteriocin showed marked changes in cellular architecture of the trophozoites with evident disorganization of the cell membrane, adhesive disc and cytoplasmic components. This is the first reported study of the safe anti-giardial efficacy of L. acidophilus (P106) derived bacteriocin, hence highlighting its great promise as a potential therapeutic safe alternative to existing commercial drugs.

Effects of Adding Barium-Borosilicate Glass to a Simplified Etch-and-rinse Adhesive on Radiopacity and Selected Properties

To evaluate the radiopacity, ultimate tensile strength (UTS), microhardness (KHN), degree of conversion (DC), water sorption (WS) and solubility (SL) of experimental adhesives. Five experimental adhesives with different concentrations of barium-borosilicate oxide microfillers [0% (R0), 30% (R30), 40% (R40), 50% (R50), 60% (R60)] were formulated based on the adhesive system Ambar (FGM). The adhesive Adper Single Bond 2 (SB, 3M ESPE) was used as commercial reference. For the radiopacity (n = 5), KHN (n = 5), WS (n = 10), and SL (n = 10) tests, adhesive disks were constructed (5.0 mm in diameter and 1.0 mm thick), while for UTS (n = 5), hourglass-shaped specimens with a cross-sectional area of 0.8 mm2 were used. The FTIR spectra of unpolymerized and polymerized adhesives were used to determine the DC. Data were submitted to a one-way ANOVA and Tukey's test (α = 0.05). All experimental adhesives showed radiopacity similar to enamel, except those of R0 and SB. Filler addition did not jeopardize the UTS, KHN, or WS of the filled adhesives in comparison with the unfilled version. Except for R40, filler addition reduced the SL. The filled adhesives showed lower DC when compared with R0, but the DC was similar or higher when compared with SB. The addition of barium-borosilicate glass up to 50% did not jeopardize the mechanical properties of the adhesive layer and seems to reduce its solubility.

Rohdella amazonica n. sp. (Aspidogastrea: Aspidogastridae) from the Amazoninan banded puffer fish Colomesus psittacus (Bloch & Schneider, 1801)

Aspidogastreans are commonly found infecting freshwater and marine molluscs, teleosts fishes and freshwater turtles. The subclass comprises four families - Rugogastridae Schell 1973, Stichocotylidae Faust & Tang 1936, Multicalycidae Gibson & Chinabut 1984 and Aspidogastridae Poche 1907 - and it is characterized by the presence of a ventral adhesive disc divided into rows of alveoli. In the current work, using light and scanning electron microscopy and molecular approaches, a new species of Aspidogastridae of the genus Rohdella Gibson & Chinabut, 1984, is described as a parasite of Colomesus psittacus in Brazil. The new taxon is distinguishable by the presence of oesophageal glands, teguments covered by ciliated papillae, and the position and shape of the hermaphroditic duct. The present work describes the third species of the genus Rohdella, thereby adding new morphological and molecular data regarding Aspidogastridae.

Mechanical properties and modeling of drug release from chlorhexidine-containing etch-and-rinse adhesives

ObjectivesTo evaluate the effects of chlorhexidine (CHX) addition in different concentrations into simplified etch-and-rinse adhesives on the ultimate tensile strength (UTS), water sorption (WS), solubility (SO) and the rate of CHX release over time. MethodsWe added CHX diacetate to Ambar [AM] (FGM) and XP Bond [XP] (Dentsply) in concentrations of 0, 0.01, 0.05, 0.1 and 0.2wt%. For UTS (n=10 for each group), adhesive specimens were constructed in an hourglass shape metallic matrix with cross-sectional area of 0.8mm2. Half of specimens were tested after 24h and the other half after 28 days of water storage in tension of 0.5mm/min. For WS and SO (n=10 for each group), adhesive discs (5.8mm×1.0mm) were prepared into a mold. After desiccation, we weighed and stored the cured adhesive specimens in distilled water for evaluation of the WS, SO and the cumulative release of CHX over a 28-day period. For CHX release (n=10 for each group), spectrophotometric measurements of storage solution were performed to examine the release kinetics of CHX. We subjected data from each test to ANOVA and Tukey’ test (α=0.05). ResultsXP Bond adhesive showed significantly more WS and SO and lower UTS than Ambar. In general, the addition of CHX did not alter WS, SO and UTS of the adhesives. XP showed a higher CHX release than AM (p<0.05) in all concentrations and the final amount of CHX release was directly proportional to the initial CHX concentration added to the adhesives. After 28 days of water storage, approximately 20% of CHX was released from XP and 8.0–12.0% from AM. ConclusionsAddition of CHX to commercial adhesive is a feasible method to provide a controlled release of CHX over time without jeopardizing WS, SO and UTS of the adhesives. SignificanceManufacturers should consider adding CHX to commercial adhesives to provide a controlled release of CHX over time.

Synthetic adhesive attachment discs inspired by spider's pyriform silk architecture

ABSTRACTSpiders attach their major ampullate silk dragline fibers to surfaces using attachment discs spun from pyriform silk fibers. These attachment discs allow spiders to move safely from place to place while secured to a dragline and to attach their webs to a variety of surfaces. Here, we demonstrate a simple electrospinning process to mimic the “staple‐pin” architecture used by spiders for these attachment discs. Experimental and theoretical evidence are provided to confirm the advantages of thousands of micron‐size “staple‐pins” and their low peeling angles to enhance the adhesive forces required to peel the natural and synthetic attachment discs. These results provide a unique architectural design for fabricating new adhesives that uses very little material for various biomedical and material science applications. © 2014 Wiley Periodicals, Inc. J Polym Sci Part B: Polym. Phys. 2014, 52, 553–560

Extraction and Characterization of Three Polysaccharides from Adhesive Discs of Parthenocissus tricuspidata

Extraction and Characterization of Three Polysaccharides from Adhesive Discs of Parthenocissus tricuspidata

Stick Tight!

The rocky intertidal zone is an extreme environment with high, variable forces from crashing waves and strong ocean currents. A family of fishes, including the northern clingfish (Gobiesox maeandricus), has evolved an adhesive disc that allows them to adhere to rocks in the intertidal zone and even launch predatory attacks on molluscs that are attached to the rocks (Figure 1). Dylan Wainwright, Thomas Kleinteich, Anja Kleinteich, Stanislav Gorb, and Adam Summers studied the morphology of this fish disc to understand the properties of a reversibly adhesive disc that has a strong tenacity to stick to irregular, slippery, and wet surfaces.

Insemination of the monogenean Neobenedenia girellae (Capsalidae, Benedeniinae)

In vitro spermatophore formation and insemination of Neobenedenia girellae (Monogenea: Capsalidae, Benedeniinae) were recorded on video and described for the first time. Upon contact of two individuals, the anterior adhesive discs of the donor firmly attached to the dorsal tegument of the recipient and the donor's fore body strongly contracted such that the genital pore region protruded and the penis was pushed anteriorly to protrude through the genital pore. It is hypothesised that the donor penis mechanically damaged the tegument of the recipient. The sperm and spermatophore matrix were released together through the penis, which was placed under the left anterior attachment disc immediately behind the adhesive pad. The spermatophore matrix containing the spermatozoa became solid and attached to the dorsal surface of recipient's body. When observed under scanning electron microscopy, the spermatophores were irregularly shaped, with a diameter of 52–83μm. Under light microscopy they consisted of a proximal eosinophilic matrix portion and a distal thin-walled portion containing spermatozoa. Both parts were enclosed with a thin outer casing. Insemination occurred during and after spermatophore formation. Three types of insemination were recorded, unilateral and mutual insemination and self-insemination. The presence of self-insemination indicates that even a single N. girellae on a cultured fish may cause a significant parasite infection in the entire aquaculture system.

An early fossil remora (Echeneoidea) reveals the evolutionary assembly of the adhesion disc

The adhesion disc of living remoras (Echeneoidea: Echeneidae) represents one of the most remarkable structural innovations within fishes. Although homology between the spinous dorsal fin of generalized acanthomorph fishes and the remora adhesion disc is widely accepted, the sequence of evolutionary-rather than developmental-transformations leading from one to the other has remained unclear. Here, we show that the early remora †Opisthomyzon (Echeneoidea: †Opisthomyzonidae), from the early Oligocene (Rupelian) of Switzerland, is a stem-group echeneid and provides unique insights into the evolutionary assembly of the unusual body plan characteristic of all living remoras. The adhesion disc of †Opisthomyzon retains ancestral features found in the spiny dorsal fins of remora outgroups, and corroborates developmental interpretations of the homology of individual skeletal components of the disc. †Opisthomyzon indicates that the adhesion disc originated in a postcranial position, and that other specializations (including the origin of pectination, subdivision of median fin spines into paired lamellae, increase in segment count and migration to a supracranial position) took place later in the evolutionary history of remoras. This phylogenetic sequence of transformation finds some parallels in the order of ontogenetic changes to the disc documented for living remoras.

Paratrichodina africana (Ciliophora: Trichodinidae) of wild and cultured Nile tilapia in the Northern Brazil

The present work morphologically characterizes Paratrichodina africana from the gills of wild and farmed Nile tilapia from Northern Brazil (eastern Amazonia). Ninety fish were captured for parasitological analysis in Macapá, State of Amapá, from a wetland area bathed by the Amazon River commonly called 'Ressaca do Zerão' (n = 52), as well as from a local fish farm (n = 38). Wet smears of the skin and gills of the captured fish were air dried at room temperature and impregnated with silver nitrate by Klein's method for posterior examination of the adhesive disc structures. Total prevalence of parasitism was 16.6% (23% in fish from wetland and 7.8% in farmed fish). Characterized as a small-sized trichodinid, it presented the following measures: 33.2 ± 4.7 µm body diameter, 17.5 ± 2.1 µm adhesive disc, 10.0 ± 0.9 µm denticulate ring, and 22.6 ± 2.0 denticles. Paratrichodina africana reported in this study strongly resembles those described for other localities, but it differs by presenting greater body length. This is the fourth report of P. africana parasitizing a host fish.