Occurrence of parasitism promoted by Myzobdella lugubris Leidy, 1851 (Annelida: Piscicolidae) in Callinectes bocourti (Crustacea: Portunidae): A report of invasive leech in Brazilian Amazon province.

This study reports the presence of high parasitic load by Myzobdella lugubris Leidy, 1851 in the swimming crab Callinectes bocourti A. Milne-Edwards, 1879 from Amazon mangrove. We sampled the swimming crabs using a baited trap, between January and June 2023, in Santa Maria River, located in the municipality of Curuçá, state of Pará, Brazil (geographical coordinates 0°40'3.705"S, 047°54'43.405"W). After sampling, each swimming crab was individually placed in plastic containers for the count of leeches per individual. In the laboratory, the specimens were sexed, measured (parasite and host) and fixed in 70% alcohol. For the leech species identification, macroscopic techniques were combined with light microscopy (LM) and scanning electron microscopy (SEM). We examined 86 specimens of C. bocourti (75 males and 11 females) in a ratio of 1 M:0.14 F, all infested with leeches. In total, 186 leech specimens were collected, ranging from 1 to 21 leeches per host. Leeches oviposited the cocoons in greater quantities in ventral area of swimming crab carapace (32%), followed by dorsal area of carapace (29.09%), chelipeds (24.34%) and ambulatory legs (14.57%). The presence of M. lugubris is a risk to the health of the host, once it may transmit a range of diseases to aquatic organisms, and subsequently risk to human health.

Biomechanics of transduction by mechanosensory cilia for prey detection in aquatic organisms.

Surface-feeding aquatic animals navigate towards the source of water disturbances and must differentiate prey from other environmental stimuli. Medicinal leeches locate prey, in part, using a distribution of mechanosensory hairs along their body that deflect under fluid flow. Leech's behavioral responses to surface wave temporal frequency are well documented. However, a surface wave's temporal frequency depends on many underlying environmental and fluid properties that vary substantially in natural habitats (e.g., water depth, temperature). The impact of these variables on neural response and behavior is unknown. Here, we developed a physics-based leech mechanosensor model to examine the impact of environmental and fluid properties on neural response. Our model used the physical properties of a leech cilium and was verified against existing behavioral and electrophysiological data. The model's peak response occurred with waves where the effects of gravity and surface tension were nearly equal (i.e., the phase velocity minimum). This suggests that preferred stimuli are related to the interaction between fundamental properties of the surrounding medium and the mechanical properties of the sensor. This interaction likely tunes the sensor to detect the nondispersive components of the signal, filtering out irrelevant ambient stimuli, and may be a general property of cilia across the animal kingdom.

'Pearls' of the nineteenth-century: from therapeutic actors to global commodities medicinal leeches in the Ottoman Empire.

Nineteenth-century physicians increasingly favoured leeching - the placing of a live leech onto a patient's skin to stimulate or limit blood flow - as a cure for numerous ailments. As conviction in their therapeutic properties spread, leech therapy dominated European medicine; France imported over fifty million leeches in one year. Demand soon outpaced supply, spawning a lucrative global trade. Over-collection and farming eventually destroyed leech habitats, wreaked environmental havoc and forced European merchants to seek new supply sources. Vast colonies of leeches were found to inhabit the immense wetlands of the Ottoman Empire, which soon became a major exporter of medicinal leeches. Following the Treaty of Balta Liman (1838), the Ottoman state moved to exert control over the lucrative trade, imposing a tax on leech gathering and contracting with tax-farmers (mültezim) to collect the taxes. British diplomats, merchants and other stakeholders protested the imposition of the tax, as had previously happened with the commodification of wildlife; their pursuit of profit led collectors and farmers to over-gather leeches, with catastrophic consequences. By the end of the century, so great had their worth climbed that the leech population faced extinction. This paper situates medicinal leeches as therapeutic actors of history and adopts an interscale approach in formulating the human-leech interaction. It offers a substantive contribution to the history of medicine, in revealing the centrality of leeches to the rise of modern medicine and global trade, but also by making visible their role in shaping imperial diplomacy and worldwide economic markets.

Motor neural networks in the leech.

Leeches display robust motor patterns and exhibit a relatively simple nervous system where neurons are unambiguously identified. This brief article focuses on Hirudo verbana and summarizes how research in this organism has contributed to insights in the field of motor control, where networks have been studied from population down to individual neuron perspectives.

Repetitive nociceptive stimulation elicits complex behavioral changes in Hirudo: evidence of arousal and motivational adaptations.

Appropriate responses to real or potential damaging stimuli to the body (nociception) are critical to an animal's short- and long-term survival. The initial goal of this study was to examine habituation of withdrawal reflexes (whole-body and local shortening) to repeated mechanical nociceptive stimuli (needle pokes) in the medicinal leech, Hirudo verbana, and assess whether injury altered habituation to these nociceptive stimuli. While repeated needle pokes did reduce shortening in H. verbana, a second set of behavior changes was observed. Specifically, animals began to evade subsequent stimuli by either hiding their posterior sucker underneath adjacent body segments or engaging in locomotion (crawling). Animals differed in terms of how quickly they adopted evasion behaviors during repeated stimulation, exhibiting a multi-modal distribution for early, intermediate and late evaders. Prior injury had a profound effect on this transition, decreasing the time frame in which animals began to carry out evasion and increasing the magnitude of these evasion behaviors (more locomotory evasion). The data indicate the presence in Hirudo of a complex and adaptive defensive arousal process to avoid noxious stimuli that is influenced by differences in internal states, prior experience with injury of the stimulated areas, and possibly learning-based processes.

Single and combined effects of cadmium, microplastics, and their mixture on whole-body serotonin and feeding behaviour following chronic exposure and subsequent recovery in the freshwater leech, Nephelopsis obscura.

Microplastics and metals are contaminants detected in many freshwater systems globally. Interactions of microplastics with other contaminants including cadmium poses potential threats to the health of aquatic organisms including Nephelopsis obscura, a predatory leech species that is widespread and serves important ecological and economic roles. The feeding biology of N. obscura has been well-described, including that serotonin regulates feeding behaviour. Further, exposure to cadmium has been found to cause decrease whole-body concentrations of serotonin. The influence that microplastic contamination and co-contamination of cadmium and microplastics has on N. obscura is unknown. The present study had three objectives: (1) to determine if water or sediment contaminated with cadmium, microplastics, or their mixture resulted in greater cadmium uptake by N. obscura, (2) to assess effects of chronic (21-day) exposure of N. obscura to waterborne cadmium, microplastics, and their mixture on bioaccumulation of cadmium, concentrations of serotonin, and feeding behaviour (latency to feeding, time spent feeding, and distance moved), and (3) to reassess the bioaccumulation of cadmium, concentrations of serotonin, and feeding behaviour following transfer to an uncontaminated environment for a one-week recovery period. This study revealed that access to and presence of sediment is protective against cadmium uptake and that cadmium is more readily accumulated from waterborne sources, even in environments where both sediment and surface water are contaminated. After 21-days of exposure to waterborne cadmium, microplastics, and their mixture, accumulation of cadmium, decreased concentrations of serotonin, and impaired feeding behaviours were greatest in leeches from the co-exposures compared to leeches from either single contaminant exposure group. Finally, after one week of depuration and recovery in freshwater following the 21-day exposures, concentrations of serotonin and feeding behaviour were restored in individuals from the microplastic exposure; however, cadmium-exposed individuals continued to show decreased concentrations of serotonin and behavioural deficits. The co-exposure of leeches to cadmium and microplastics resulted in additive effects to serotonin synthesis and feeding behaviour; however, this study demonstrated that leeches were able to recover from microplastic toxicity within a week whereas cadmium toxicity persisted.

Effects of mammalian blood with different glucose levels on reproduction, growth and survival of the southern medicinal leech, Hirudo verbana Carena, 1820.

Medicinal leeches are generally fed using pure mammalian blood. In the present study reproduction, growth and survival of medicinal leeches (Hirudo spp.) fed by mammalian blood with modified glucose level were investigated for the first time. Leeches were fed by cattle blood in a final glucose level of 152 mg/dL (control group; Glucose-free), 200 mg/dL (G200 group), 300 mg/dL (G300 group), 500 mg/dL (G500 group), 750 mg/dL (G750 group), 1000 mg/dL (G1000 group), 2500 mg/dL (G2500 group) and 5000 mg/dL (G5000 group) with the addition of D-Glucose Monohydrate. Greatest growth performance was determined in the G2500 group with a specific growth rate of 2.34% (final body weight: 10.37 ± 3.86 g) (P < 0.05). A quadratic increase was observed in the body weight values of the leeches depending on the glucose dose (Plinear and Pquadratic < 0.05). The greatest survival and gravidity rates were 89% and 38%, respectively, in the G750 group (P < 0.05). The increased glucose level caused a sharp decrease in the survival and gravidity rates of leeches. The glucose level did not significantly effected the cocoon and offspring productivity (P > 0.05). According to the broken line model, optimum glucose levels based on growth, survival rate and gravidity rate were 2461 mg/dL, 750.0 mg/dL and 749.9 mg/dL, respectively. The study showed that, although the optimum growth performance was obtained in the G2500 group, blood with glucose level of 750 mg/dL should be used for profitable medicinal leech culture considering survival and gravidity rates.

Serotonin mediates stress-like effects on responses to non-nociceptive stimuli in the medicinal leech Hirudo verbana.

Noxious stimuli can elicit stress in animals that produce a variety of adaptations including changes in responses to nociceptive and non-nociceptive sensory input. One example is stress-induced analgesia that may be mediated, in part, by the endocannabinoid system. However, endocannabinoids can also have pro-nociceptive effects. In this study, the effects of electroshock, one experimental approach for producing acute stress, were examined on responses to non-nociceptive mechanical stimuli and nociceptive thermal stimuli in the medicinal leech (Hirudo verbana). The electroshock stimuli did not alter the leeches' responses to nociceptive stimuli, but did cause sensitization to non-nociceptive stimuli, characterized by a reduction in response threshold. These experiments were repeated with drugs that either blocked synthesis of the endocannabinoid transmitter 2-arachidonoylglycerol (2-AG) or transient receptor potential vanilloid (TRPV) channel, which is known to act as an endocannabinoid receptor. Surprisingly, neither treatment had any effect on responses following electroshock. However, the electroshock stimuli reliably increased serotonin (5-hydroxytryptamine or 5HT) levels in the H. verbana CNS. Injection of 5HT mimicked the effects of the electroshocks, sensitizing responses to non-nociceptive stimuli and having no effect on responses to nociceptive stimuli. Injections of the 5HT receptor antagonist methysergide reduced the sensitization effect to non-nociceptive stimuli after electroshock treatment. These results indicate that electroshocks enhance response to non-nociceptive stimuli but do not alter responses to nociceptive stimuli. Furthermore, while 5HT appears to play a critical role in this shock-induced sensitizing effect, the endocannabinoid system seems to have no effect.

Intersegmental Interactions Give Rise to a Global Network.

Animal motor behaviors require the coordination of different body segments. Thus the activity of the networks that control each segment, which are distributed along the nerve cord, should be adequately matched in time. This temporal organization may depend on signals originated in the brain, the periphery or other segments. Here we evaluate the role of intersegmental interactions. Because of the relatively regular anatomy of leeches, the study of intersegmental coordination in these animals restricts the analysis to interactions among iterated units. We focused on crawling, a rhythmic locomotive behavior through which leeches move on solid ground. The motor pattern was studied ex vivo, in isolated ganglia and chains of three ganglia, and in vivo. Fictive crawling ex vivo (crawling) displayed rhythmic characteristics similar to those observed in vivo. Within the three-ganglion chains the motor output presented an anterior-posterior order, revealing the existence of a coordination mechanism that occurred in the absence of brain or peripheral signals. An experimental perturbation that reversibly abolished the motor pattern in isolated ganglia produced only a marginal effect on the motor activity recorded in three-ganglion chains. Therefore, the segmental central pattern generators present in each ganglion of the chain lost the autonomy observed in isolated ganglia, and constituted a global network that reduced the degrees of freedom of the system. However, the intersegmental phase lag in the three-ganglion chains was markedly longer than in vivo. This work suggests that intersegmental interactions operate as a backbone of correlated motor activity, but additional signals are required to enhance and speed coordination in the animal.

Approaches to studying injury-induced sensitization and the potential role of an endocannabinoid transmitter.

Endocannabinoids are traditionally thought to have an analgesic effect. However, it has been shown that while endocannabinoids can depress nociceptive signaling, they can also enhance non-nociceptive signaling. Therefore, endocannabinoids have the potential to contribute to non-nociceptive sensitization after an injury. Using Hirudo verbana (the medicinal leech), a model of injury-induced sensitization was developed in which a reproducible piercing injury was delivered to the posterior sucker of Hirudo. Injury-induced changes in the non-nociceptive threshold of Hirudo were determined through testing with Von Frey filaments and changes in the response to nociceptive stimuli were tested by measuring the latency to withdraw to a nociceptive thermal stimulus (Hargreaves apparatus). To test the potential role of endocannabinoids in mediating injury-induced sensitization, animals were injected with tetrahydrolipstatin (THL), which inhibits synthesis of the endocannabinoid transmitter 2-arachidonoylglycerol (2-AG). Following injury, a significant decrease in the non-nociceptive response threshold (consistent with non-nociceptive sensitization) and a significant decrease in the response latency to nociceptive stimulation (consistent with nociceptive sensitization) were observed. In animals injected with THL, a decrease in non-nociceptive sensitization in injured animals was observed, but no effect on nociceptive sensitization was observed.

Reproduction of Limnatis paluda (Tennent 1859) in laboratory conditions.

The reproductive functions of Limnatis paluda were studied in laboratory conditions including imposing of two thermal conditions (T1: 18 ± 2 °C and T2: 26 ± 2 °C) and with there being placement of three bedding substrates (tile, moss, and stone) in the containers specimens were confined. Furthermore, values for reproductive variables of cocoons and hatchlings were recorded. With the T2 treatment, gravid adults had cocoon depositions, whereas those when there was imposing of the T1 treatment did not have cocoon depositions. There was cocoon deposition only on the moss bedding, and it is presumed that moss could provide a robust and stable nursing microenvironment for developing embryos. Frequency (%) of cocoon depositions did not vary during the months of the deposition period. Hatching occurred after 26 ± 2 days when there was imposing of the T2 treatment. There was no correlation between the values for size of the ovigerous cocoons and number of offspring. There was a negative correlation between number and mean weight of hatchlings. There was a difference in cocoon weight for the different months and fluctuation in mortality and hatching rate which is assumed to be related to differences in feeding behaviors.

Function of the waterfowl nests as reproduction and living areas for leeches (Annelida: Hirudinea).

Some species of leeches migrate into waterfowl nests and use these both as general habitats and to deposit cocoons, but ecological associations between leeches and birds are not well understood. In the present study, characteristics of waterfowl nests both as living area for leeches (Hirudinea) and as reproduction areas for select hirudinid leeches (Hirudo verbana and Haemopis sanguisuga) was investigated in both natural and controlled laboratory conditions. A total of 48 leeches were detected in 23 of the 51 nests surveyed in natural habitats. The leeches were detected more frequently but there were less dense populations in the nests of the great crested grebe (Podiceps cristatus) (frequency: 57 %, population density: 0.71 ± 0.76 leeches/nest), and leeches were detected less frequently but there were more dense populations in the nests of coots (Fulica atra) (frequency: 36 %, population density: 0.91 ± 1.70 leeches/nest). Although the hirudinid leeches naturally reproduce between June and September, cocoons were only detected in August and September, when the nests were not actively used by the waterfowl and when water depths were less. In laboratory conditions, gravid medicinal leeches prefer moist peat rather than waterfowl nests for cocoon deposition. Results from the present study indicate leeches more frequently use the nests of coots, which have both dry and wet layers, compared to the nests of the great crested grebe, which have only a wet layer. Leeches were also found to be more prevalent in bird nests during spring months, during the reproductive periods of the waterfowl.

Morphological mechanism allowing a parasitic leech, Ozobranchus jantseanus (Rhynchobdellida: Ozobranchidae), to survive in ultra-low temperatures.

Ozobranchus jantseanus is the largest metazoan known to survive in liquid nitrogen without pretreatment to date; however, the mechanism underlying this tolerance remains unclear. In this study, the first analyses of histological and morphological changes in normal, frozen, and dehydrated states were performed. Adults survived after direct placement in liquid nitrogen for 96 h, with a survival rate of approximately 86.7%. The leech could withstand rapid desiccation and its survival rate after rehydration was 100% when its water loss was below about 84.8%. After freezing, desiccation, and ethanol dehydration, the leech immediately formed a hemispherical shape. Particularly during drying, an obvious transparent glass-like substance was observed on surface. Scanning electron microscopy revealed many pores on the surface of the posterior sucker, creating a sponge-like structure, which may help to rapidly expel water, and a hemispherical shape may protect the internal organs by contraction and folding reconstruction in the anterior-posterior direction. A substantial amount of mucopolysaccharides on the surface and acid cells and collagen fibers in the body, all of which contained substantial polysaccharides, may play a key protective role during freezing. Our results indicate that the resistance of leeches to ultra-low temperatures can be explained by cryoprotective dehydration/vitrification strategies. This article has an associated First Person interview with the first author of the paper.

Behavioral variation according to feeding organ diversification in glossiphoniid leeches (Phylum: Annelida).

Adaptive radiation is a phenomenon in which various organs are diversified morphologically or functionally as animals adapt to environmental inputs. Leeches exhibit a variety of ingestion behaviors and morphologically diverse ingestion organs. In this study, we investigated the correlation between behavioral pattern and feeding organ structure of leech species. Among them, we found that Alboglossiphonia sp. swallows prey whole using its proboscis, whereas other leeches exhibit typical fluid-sucking behavior. To address whether the different feeding behaviors are intrinsic, we investigated the behavioral patterns and muscle arrangements in the earlier developmental stage of glossiphoniid leeches. Juvenile Glossiphoniidae including the Alboglossiphonia sp. exhibit the fluid ingestion behavior and have the proboscis with the compartmentalized muscle layers. This study provides the characteristics of leeches with specific ingestion behaviors, and a comparison of structural differences that serves as the first evidence of the proboscis diversification.

Myxobdella socotrensis sp. nov., a new parasitic leech from Socotra Island, with comments on the phylogeny of Praobdellidae (Hirudinida: Arhynchobdellida).

The Socotra Archipelago (Yemen) is remarkable for unique ecosystems and high endemism, for which it is often referred to as the "Galápagos of the Indian Ocean". Here we describe a new parasitic leech Myxobdella socotrensis sp. nov. from Socotra, the largest island of the archipelago. The new species was found in a freshwater spring attached to the endemic crab Socotrapotamon socotrensis (Hilgendorf, 1883) (Crustacea: Potamidae). Based on its morphology, ecology and a phylogenetic analysis of DNA sequence data (12S, 18S, 28S and COI gene markers), the new leech species is classified into the highly diversified family Praobdellidae (Hirudinida: Arhynchobdellida), distributed in tropical regions of Africa, Asia and Central and South America. The praobdellid leeches are known to infest mammalian mucous membranes, however, some taxa are associated with other hosts, namely crabs, amphibians and birds. By its morphology, the new species fits quite well in the current concept of the genus Myxobdella Oka, 1917. However, the monophyly of Myxobdella was not supported here by molecular data, pointing at the need of a more comprehensive systematic revision of the genus and family. As far as known, Myxobdella socotrensis sp. nov. is endemic to the Socotra Island, but more data are needed to understand its evolutionary origin, biology and distribution.

Anatomy and activity patterns in a multifunctional motor neuron and its surrounding circuits.

Dorsal Excitor motor neuron DE-3 in the medicinal leech plays three very different dynamical roles in three different behaviors. Without rewiring its anatomical connectivity, how can a motor neuron dynamically switch roles to play appropriate roles in various behaviors? We previously used voltage-sensitive dye imaging to record from DE-3 and most other neurons in the leech segmental ganglion during (fictive) swimming, crawling, and local-bend escape (Tomina and Wagenaar, 2017). Here, we repeated that experiment, then re-imaged the same ganglion using serial blockface electron microscopy and traced DE-3's processes. Further, we traced back the processes of DE-3's presynaptic partners to their respective somata. This allowed us to analyze the relationship between circuit anatomy and the activity patterns it sustains. We found that input synapses important for all the behaviors were widely distributed over DE-3's branches, yet that functional clusters were different during (fictive) swimming vs. crawling.

The Vascularity of Ayurvedic Leech Therapy: Sensory Translations and Emergent Agencies in Interspecies Medicine.

This article offers vascularity as a multi-dimensional imaginary for the interspecies entanglements constituting Ayurvedic leech therapy. Whether, when, where, and how a leech decides to bite, suck, and release comprise pivotal junctures in leech therapy as practiced in southern Kerala, India. In the course of leech-human intra-actions, leeches translate matter, providing sensory mediation, relief, and amusement. Enmeshed in social and ecological relations inflected by gender, religion, class, and caste, this analysis of Ayurvedic leech therapy reframes questions of agencies starting with and from the viewpoint of the vascular capacities of leeches in their interactions with humans. This image of vascularity provides an analytic for the emergent agencies of humans and leeches constituted by sensory intra-actions at branching points in this multispecies clinical practice.

The life cycle of the Siamese shield leech, Placobdelloides siamensis Oka, 1917.

Placobdelloides siamensis is a glossiphoniid leech with a short life cycle. In a laboratory setting, ten mature P. siamensis left their host (a turtle) after feeding for approximately three days and initiated copulation. The adults spent 3-4 days gestating before depositing eggs (272.8±62.9 eggs/clutch; range: 186-359 eggs/clutch). The eggs then changed from a creamy white to a creamy brownish color before hatching. Hatching occurred after incubation on the parent's ventral surface for 5-7 days. The transparent brood, with a single pair of red eyes, spent a couple of weeks under the venters of their parents. After this period, they left their parents and grew to maturity in 10-15 days; leeches were considered mature when their color was similar to that of their parents and they performed their first copulation. In addition, the mature leeches survived for 163 days on one feeding.

A tale of two leeches: Toward the understanding of the evolution and development of behavioral neural circuits.

In the animal kingdom, behavioral traits encompass a broad spectrum of biological phenotypes that have critical roles in adaptive evolution, but an EvoDevo approach has not been broadly used to study behavior evolution. Here, we propose that, by integrating two leech model systems, each of which has already attained some success in its respective field, it is possible to take on behavioral traits with an EvoDevo approach. We first identify the developmental changes that may theoretically lead to behavioral evolution and explain why an EvoDevo study of behavior is challenging. Next, we discuss the pros and cons of the two leech model species, Hirudo, a classic model for invertebrate neurobiology, and Helobdella, an emerging model for clitellate developmental biology, as models for behavioral EvoDevo research. Given the limitations of each leech system, neither is particularly strong for behavioral EvoDevo. However, the two leech systems are complementary in their technical accessibilities, and they do exhibit some behavioral similarities and differences. By studying them in parallel and together with additional leech species such as Haementeria, it is possible to explore the different levels of behavioral development and evolution.