Dynamics of actinotrichia regeneration in the adult zebrafish fin.

The skeleton of adult zebrafish fins comprises lepidotrichia, which are dermal bones of the rays, and actinotrichia, which are non-mineralized spicules at the distal margin of the appendage. Little is known about the regenerative dynamics of the actinotrichia-specific structural proteins called Actinodins. Here, we used immunofluorescence analysis to determine the contribution of two paralogous Actinodin proteins, And1/2, in regenerating fins. Both proteins were detected in the secretory organelles in the mesenchymal cells of the blastema, but only And1 was detected in the epithelial cells of the wound epithelium. The analysis of whole mount fins throughout the entire regenerative process and longitudinal sections revealed that And1-positive fibers are complementary to the lepidotrichia. The analysis of another longfin fish, a gain-of-function mutation in the potassium channel kcnk5b, revealed that the long-fin phenotype is associated with an extended size of actinotrichia during homeostasis and regeneration. Finally, we investigated the role of several signaling pathways in actinotrichia formation and maintenance. This revealed that the pulse-inhibition of either TGFß/Activin-ßA or FGF are sufficient to impair deposition of Actinodin during regeneration. Thus, the dynamic turnover of Actinodin during fin regeneration is regulated by multiple factors, including the osteoblasts, growth rate in a potassium channel mutant, and instructive signaling networks between the epithelium and the blastema of the regenerating fin.

Ultrastructural changes to the tegumental system and gastrodermal cells of adult Fasciola hepatica following treatment in vivo with a commercial preparation of myrrh (Mirazid).

An in vivo study in the laboratory rat model has been carried out to monitor changes to the tegument and gut of adult Fasciola hepatica following treatment with myrrh ('Mirazid'). Rats infected with the triclabendazole-resistant Dutch isolate were dosed orally with Mirazid at a concentration of 250 mg/kg and flukes recovered 2, 3 and 7 days post-treatment (pt). The flukes were processed for examination by scanning and transmission electron microscopy. A variety of changes to the external surface were observed, culminating in the sloughing of the tegumental syncytium. Internal changes to the syncytium and tegumental cell bodies were more severe and were evident from 2 days pt onwards. Swelling of the basal infolds (leading to flooding of the surface layer) and a decline in secretory body production were the major changes seen. The gastrodermal cells were less severely affected than the tegument, pointing to a trans-tegumental route of uptake for Mirazid by the fluke. Some loss of muscle fibres in the main somatic muscle layers was observed, which may be correlated with the decline in movement of flukes seen at recovery.

Structural and Developmental Disparity in the Tentacles of the Moon Jellyfish Aurelia sp.1.

Tentacles armed with stinging cells (cnidocytes) are a defining trait of the cnidarians, a phylum that includes sea anemones, corals, jellyfish, and hydras. While cnidarian tentacles are generally characterized as structures evolved for feeding and defense, significant variation exists between the tentacles of different species, and within the same species across different life stages and/or body regions. Such diversity suggests cryptic distinctions exist in tentacle function. In this paper, we use confocal and transmission electron microscopy to contrast the structure and development of tentacles in the moon jellyfish, Aurelia species 1. We show that polyp oral tentacles and medusa marginal tentacles display markedly different cellular and muscular architecture, as well as distinct patterns of cellular proliferation during growth. Many structural differences between these tentacle types may reflect biomechanical solutions to different feeding strategies, although further work would be required for a precise mechanistic understanding. However, differences in cell proliferation dynamics suggests that the two tentacle forms lack a conserved mechanism of development, challenging the textbook-notion that cnidarian tentacles can be homologized into a conserved bauplan.

In vitro screening for cestocidal activity of three species of Cassia plants against the tapeworm Raillietina tetragona.

Different species of Cassia plant are widely available in India and are commonly used either for their laxative, antimicrobial or antibacterial activity. In the present study the effectiveness in vitro of the crude alcoholic extracts of three species, namely Cassia alata, C. occidentalis and C. angustifolia, in the early paralysis and mortality of the fowl tapeworm Raillietina tetragona at concentrations ranging from 5 to 80 mg/ml was investigated. Time of paralysis and death were monitored frequently. Immediately after paralysis the tapeworms were processed for electron microscopic studies. While the untreated or control parasites survived for 81.93 ± 5.85 h, the parasites treated with C. alata took less time (1.68 ± 0.27 h) to be paralysed, followed by those treated with C. angustifolia (2.95 ± 0.29 h). Although C. occidentalis took more time (4.13 ± 0.31 h) to paralyse, in combination with either C. alata or C. angustifolia the time taken to paralyse became shorter. All the plant-treated parasites showed irrevocable changes in the scolex and proglottids as compared with the control, and these observations are comparable with those obtained with praziquantel. These results indicate that the three plants tested can be claimed to have anthelmintic activity in addition to their known properties, both when used individually and in combination. Further investigations will be required to evaluate their mechanism of action.

Toxicological and histopathological effects of hydramethylnon on Atta sexdens rubropilosa (Hymenoptera: Formicidae) workers.

The leaf-cut ants are important agricultural pest, because they can cause intense defoliation in plants and destroy large areas cultivated. Although there are several works for the control of these insects by examining the toxicity of natural chemical compounds on various species of ants, few are focused on analyses of morphological changes caused in the affected organs. The aim of this study was to evaluate the effects of hydramethylnon on Atta sexdens rubropilosa workers through toxicological bioassays and morphological analysis of the post-pharyngeal glands, midgut, and Malpighian tubules of these ants. Hydramethylnon dissolved either in acetone (HA) or in a mixture of acetone and soy oil (HAO) was added to the artificial diet at a concentration of 200µg/mL. The workers fed daily with the diet containing hydramethylnon showed higher mortality than the controls, especially when HAO was used. Moreover, light and electron microscopy revealed morphological alterations in the midgut and Malpighian tubules of workers treated with HA, whereas alterations of the post-pharyngeal glands were observed in the HAO-treated group. These results indicated that the presence of soy oil provided an alternate route for the ingestion of the formicide's active ingredient and corroborated previous studies that suggested a role for the post-pharyngeal glands in lipid metabolism. Our findings suggest that the oil may carry hydramethylnon to the gland lumen, resulting in lower quantity of the active ingredient in the intestinal lumen and Malpighian tubules that explains the lower degree of morphological alterations in these structures in the workers treated with HAO. These results may provide insight into the toxicological effects of hydramethylnon on leaf-cutting ants and the use of vegetable oil as an adjuvant in baits to control ants.

The stomatopod dactyl club: a formidable damage-tolerant biological hammer.

Nature has evolved efficient strategies to synthesize complex mineralized structures that exhibit exceptional damage tolerance. One such example is found in the hypermineralized hammer-like dactyl clubs of the stomatopods, a group of highly aggressive marine crustaceans. The dactyl clubs from one species, Odontodactylus scyllarus, exhibit an impressive set of characteristics adapted for surviving high-velocity impacts on the heavily mineralized prey on which they feed. Consisting of a multiphase composite of oriented crystalline hydroxyapatite and amorphous calcium phosphate and carbonate, in conjunction with a highly expanded helicoidal organization of the fibrillar chitinous organic matrix, these structures display several effective lines of defense against catastrophic failure during repetitive high-energy loading events.

Parathyroid hormone 1 (1-34) acts on the scales and involves calcium metabolism in goldfish.

The effect of fugu parathyroid hormone 1 (fugu PTH1) on osteoblasts and osteoclasts in teleosts was examined with an assay system using teleost scale and the following markers: alkaline phosphatase (ALP) for osteoblasts and tartrate-resistant acid phosphatase (TRAP) for osteoclasts. Synthetic fugu PTH1 (1-34) (100pg/ml-10ng/ml) significantly increased ALP activity at 6h of incubation. High-dose (10ng/ml) fugu PTH1 significantly increased ALP activity even after 18h of incubation. In the case of TRAP activity, fugu PTH1 did not change at 6h of incubation, but fugu PTH1 (100pg/ml-10ng/ml) significantly increased TRAP activity at 18h. Similar results were obtained for human PTH (1-34), but there was an even greater response with fugu PTH1 than with human PTH. In vitro, we demonstrated that both the receptor activator of the NF-κB ligand in osteoblasts and the receptor activator NF-κB mRNA expression in osteoclasts increased significantly by fugu PTH1 treatment. In an in vivo experiment, fugu PTH1 induced hypercalcemia resulted from the increase of both osteoblastic and osteoclastic activities in the scale as well as the decrease of scale calcium contents after fugu PTH1 injection. In addition, an in vitro experiment with intramuscular autotransplanted scale indicated that the ratio of multinucleated osteoclasts/mononucleated osteoclasts in PTH-treated scales was significantly higher than that in the control scales. Thus, we concluded that PTH acts on osteoblasts and osteoclasts in the scales and regulates calcium metabolism in goldfish.

Mechanical response of the tympanal membranes of the tree cricket Oecanthus henryi.

Crickets have two tympanal membranes on the tibiae of each foreleg. Among several field cricket species of the genus Gryllus (Gryllinae), the posterior tympanal membrane (PTM) is significantly larger than the anterior membrane (ATM). Laser Doppler vibrometric measurements have shown that the smaller ATM does not respond as much as the PTM to sound. Hence the PTM has been suggested to be the principal tympanal acoustic input to the auditory organ. In tree crickets (Oecanthinae), the ATM is slightly larger than the PTM. Both membranes are structurally complex, presenting a series of transverse folds on their surface, which are more pronounced on the ATM than on the PTM. The mechanical response of both membranes to acoustic stimulation was investigated using microscanning laser Doppler vibrometry. Only a small portion of the membrane surface deflects in response to sound. Both membranes exhibit similar frequency responses, and move out of phase with each other, producing compressions and rarefactions of the tracheal volume backing the tympanum. Therefore, unlike field crickets, tree crickets may have four instead of two functional tympanal membranes. This is interesting in the context of the outstanding question of the role of spiracular inputs in the auditory system of tree crickets.

Dynamic range compression in the honey bee auditory system toward waggle dance sounds.

Honey bee foragers use a "waggle dance" to inform nestmates about direction and distance to locations of attractive food. The sound and air flows generated by dancer's wing and abdominal vibrations have been implicated as important cues, but the decoding mechanisms for these dance messages are poorly understood. To understand the neural mechanisms of honey bee dance communication, we analyzed the anatomy of antenna and Johnston's organ (JO) in the pedicel of the antenna, as well as the mechanical and neural response characteristics of antenna and JO to acoustic stimuli, respectively. The honey bee JO consists of about 300-320 scolopidia connected with about 48 cuticular "knobs" around the circumference of the pedicel. Each scolopidium contains bipolar sensory neurons with both type I and II cilia. The mechanical sensitivities of the antennal flagellum are specifically high in response to low but not high intensity stimuli of 265-350 Hz frequencies. The structural characteristics of antenna but not JO neurons seem to be responsible for the non-linear responses of the flagellum in contrast to mosquito and fruit fly. The honey bee flagellum is a sensitive movement detector responding to 20 nm tip displacement, which is comparable to female mosquito. Furthermore, the JO neurons have the ability to preserve both frequency and temporal information of acoustic stimuli including the "waggle dance" sound. Intriguingly, the response of JO neurons was found to be age-dependent, demonstrating that the dance communication is only possible between aged foragers. These results suggest that the matured honey bee antennae and JO neurons are best tuned to detect 250-300 Hz sound generated during "waggle dance" from the distance in a dark hive, and that sufficient responses of the JO neurons are obtained by reducing the mechanical sensitivity of the flagellum in a near-field of dancer. This nonlinear effect brings about dynamic range compression in the honey bee auditory system.

Exploring gradients of halogens and zinc in the surface and subsurface of Nereis jaws.

The outstanding mechanical properties of impact-bearing tissues, such as Nereis jaws, make their morphology and chemical composition a subject of particular interest. The complex structure of the jaw was recently reported to exhibit molecular gradients that were closely correlated with stiffness and hardness.(18) Accordingly, we have explored the spatial distribution and bonding chemistries of Zn and the halogens in the surface structure of the jaws. Using secondary ion mass spectrometry (SIMS) and scanning electron microscopy (SEM), we found that Cl, Br, and I distributions are enhanced in surface layers of the basal protected portion of the jaw but are shifted to greater depths toward the exposed jaw tip. There are thus two complementary halogen gradients in the jaw: one on the surface that decreases from the base to the tip, coupled to an increasing one in the subsurface layers. The outer surface coating appeared to have granular morphology, in contrast to the anisotropic, fibrous core that dominates the subarchitecture. Using X-ray photoelectron spectroscopy (XPS), we discovered that Zn, I, and Br in the jaws have single chemical environments whereas chlorine is present in two distinct modes (Cl-Zn and Cl-C). Given the inverse relationship between surface exposure and halogen abundance in the jaws, it is unlikely that the halogens contribute directly to mechanical properties such as wear and hardness.

The sensilla of Aedes and Anopheles mosquitoes and their importance in repellency.

The aim of this study was to detect the role of some mosquito organs in their sensation of repellent materials. A total of 250 females (15 days old) of the target species Aedes aegypti and Anopheles stephensi were prepared and divided into five groups: group 1, without antenna; group 2, without maxillary bulbs; group 3, without proboscis; group 4, without frontal tarsus; and group 5, normal females as control. A mixture of five oils containing Litsea cubeba 1%, Melaleuca leucadendron 1%, Melaleuca quinquenervia 1%, Viola odorata 1%, and Nepeta cataria 1% was included in a complex solvent containing 20% genapol, 10% polyethylene glycol, 20% ethanol, and 50% water. Furthermore, Bayrepel was used in this experiment at a 20% concentration in the same solvent. Pure water was used as control in this study. The test was carried out by spreading 100 microl of the repellent material or water on a 30-cm2 exposure area of a human volunteer's arm. In A. aegypti, the biting and landing percentages increased significantly in those mosquito groups that lacked some organs (especially maxillary bulbs), while in A. stephensi, it became not clear which organ is responsible for perception of repellents.