Observations on parasitism in deep-sea hydrothermal vent and seep limpets

Parasite burdens of shallow-water molluscs have been well documented, but little is known about parasite burdens of molluscs from deep-sea chemosynthetic environments (e.g. hydrothermal vents and seeps). Chemosynthetic habitats are characterized by high concentrations of reduced sulfur and, in the case of vents, high heavy metal concentrations. These compounds are noxious and even stress-inducing in some environments, but are part of the natural chemical milieu of vents and seeps. To examine parasite types and infection intensities in limpets from vents and seeps we documented parasite burdens in 4 limpet species from 4 hydrothermal vent fields (3 on the East Pacific Rise, 1 on the Mid-Atlantic Ridge) and 1 seep site (Florida Escarpment). Approximately 50 % of all limpets examined were infected with 1 or more types of parasites. Limpet parasites were predominantly rickettsia-like inclusions in the digestive and gill epithelia. Limpets collected from the vent field on the Mid-Atlantic Ridge were free of parasites. We detected no histopathological effects that we could attribute to parasites.     

The anatomy of Lepetella sierrai (Vetigastropoda,Lepetelloidea): implications for reproduction,feeding, and symbiosis in lepetellid limpets

The Lepetelloidea, a clade of small limpet‐shaped gastropods, can be used as a case study in continental margin and deep‐sea diversification. Lineages in this clade have been found associated with a combination of different substrates, including hydrothermal vents, seeps, wood, whale carcasses, polychaete tubes, chondrichthyan egg cases, seagrass rhizomes, algal holdfasts, crab carapaces, and sponges. Members of one lepetelloidean family, Lepetellidae, live on or inside empty tubes of members of the polychaete genus Hyalinoecia. The detailed morphology of a Mediterranean species, Lepetella sierrai Dantart & Luque 1944, was reconstructed in three dimensions from serial semi‐thin sections and compared with that of eleven other members of Lepetellidae. The hermaphroditic lepetellid limpets possessed a ciliated seminal groove, distinct testis and ovary with a common distal gonoduct, and a seminal receptacle containing mature sperm. A unique alimentary tract, with huge esophageal pouches, no true stomach, an extensive multilobed midgut, and short intestine, was present. Additionally, a bacteriocyte system throughout the entire mantle rim was revealed via light and transmission electron microscopy. This is the first recognized evidence for intracellular microbial symbiosis in lepetelloidean limpets. Semi‐thin sections showed evidence of a parasite, potentially a chitonophilid copepod, penetrating the body wall of the limpet. Hypotheses about reproductive biology, feeding, and symbiosis are presented based on anatomical features and knowledge of the habitat described herein.     

Phylogenetic characterization of episymbiotic bacteria hosted by a hydrothermal vent limpet (lepetodrilidae, vetigastropoda)

Marine invertebrates hosting chemosynthetic bacterial symbionts are known from multiple phyla and represent remarkable diversity in form and function. The deep-sea hydrothermal vent limpet Lepetodrilus fucensis from the Juan de Fuca Ridge complex hosts a gill symbiosis of particular interest because it displays a morphology unique among molluscs: filamentous bacteria are found partially embedded in the host's gill epithelium and extend into the fluids circulating across the lamellae. Our objective was to investigate the phylogenetic affiliation of the limpet's primary gill symbionts for comparison with previously characterized bacteria. Comparative 16S rRNA sequence analysis identified one γ- and three ε-Proteobacteria as candidate symbionts. We used fluorescence in situ hybridization (FISH) to test which of these four candidates occur with the limpet's symbiotic gill bacteria. The γ-proteobacterial probes consistently hybridized to the entire area where symbiotic bacteria were found, but fluorescence signal from the ε-proteobacterial probes was generally absent. Amplification of the γ-proteobacterial 16S rRNA gene using a specific forward primer yielded a sequence similar to that of limpets collected from different ridge sections. In total, direct amplification or FISH identified a single γ-proteobacterial lineage from the gills of 23 specimens from vents separated by a distance up to about 200 km and collected over the course of 2 years, suggesting a highly specific and widespread symbiosis. Thus, we report the first filamentous γ-proteobacterial gill symbiont hosted by a mollusc.     

The subcutaneous attachment of the monogenean Heterobothrium elongatum (Diclidophoridae) in the gills of Torquigener pleurogramma (Pisces: Tetraodontidae)

Heterobothrium elongatum predominantly occurred on the most anterior pair of gill arches of infected pufferfish, Torquigener pleurogramma. Juvenile and immature stages attached to gill lamellae and were concentrated on the ventral regions of the anterior pair of gill arches. Following penetration of subcutaneous tissues, young adult worms formed a subcutaneous channel either directly above or below the transverse septum, a tissue layer connecting the bases of adjacent gill rays. Large, sexually mature worms extended for most or all of the length of the gill arch where they filled the area normally occupied by the longitudinal lymphatic vessel (branchial vein) or lay adjacent to the efferent branchial and colateral arteries. Subcutaneous attachment by the parasite induced a marked inflammatory reaction in the host. During growth, the mid-region of the parasite became markedly elongate and formed a stalk connecting the haptor lying in the dorsal region of the gill and the body which emerged from the lower margin of the gill where penetration began.     

Differential precocious sexual development of Proctoeces lintoni (Digenea: Fellodistomidae) in three sympatric species of keyhole limpets Fissurella spp. may affect transmission to the final host.

The prevalence, abundance, and developmental status of the digenetic trematode Proctoeces lintoni Siddiqui et Cable 1960 were compared in 3 species of keyhole limpets Fissurella. A total of 197 limpets was collected at Caleta Chome, south-central Chile. Fissurella picta and F. costata had the highest prevalence of infection, whereas F. picta showed the greatest abundance of parasites, which increased with host shell length. However, the frequency of P. lintoni specimens with eggs in the uterus was greatest in F. costata. These results suggest that an increased rate of development of a parasite in the intermediate host may shorten the residence time necessary for maturation in the final host. Thus, faster development of the parasite in F. costata suggests the possibility that the parasites transmitted through this host species have shorter maturation times in clingfishes than individuals transmitted via other limpet species.     

Epizootiological studies of Amblyospora camposi (Microsporidia: Amblyosporidae) in Culex renatoi (Diptera: Culicidae) and Paracyclops fimbriatus fimbriatus (Copepoda: Cyclopidae) in a bromeliad habitat

The epizootiology of Amblyospora camposi was studied in a natural population of Culex renatoi, a bromeliad-inhabiting mosquito, and its intermediate host, Paracyclops fimbriatus fimbriatus, over a 2-year period. Twenty Eryngium cabrerae plants were sampled monthly from January 2003 to January 2005 and the prevalence of A. camposi in P.f. fimbriatus and Cx. renatoi populations was determined. The monthly prevalence rates of meiospore infections in Cx. renatoi larvae never exceeded 5.5% and was detected in 50% of the monthly samples. Meiospores were available in plants over the course of the study at a mean concentration of 2 x 10(4) meiospores/ml. Within each plant the parasite was maintained by horizontal transmission. P.f. fimbriatus with vegetative stages and mature spores were found regularly in bromeliads suggesting efficient meiospore infectivity to field copepod populations. The mean concentration of spores from copepods found in plants was 8 x 10(2) spores/ml. Infections in copepods were detected in 54% of the monthly samples with a prevalence rate ranging from 0.55 to 17.4% and an overall average of 5.1%. Vegetative stages in fourth instar mosquito larvae (probably derived from the horizontal pathway via spores formed in copepods) were detected in 12.5% of the monthly samples with an overall prevalence rate of 1.1%. Infections in female and male adults were detected in 20.8% of the monthly samples with an overall average of 4.1% and 6.8%, respectively.     

Epizootiology of Amblyospora stimuli (Microsporidiida: Amblyosporidae) infections in field populations of a univoltine mosquito, Aedes stimulans (Diptera: Culicidae), inhabiting a temporary vernal pool.

The epizootiology of the microsporidium Amblyospora stimuli was studied in natural populations of a univoltine mosquito, Aedes stimulans, inhabiting a temporary vernal pool over an 18-year period. The yearly prevalence of benign oenocytic infections in adult females was variable, ranging from 1.0 to 9.6% (mean = 5.1%). The yearly prevalence of transovarially transmitted meiospore infections in larval populations was consistently lower but less variable, ranging from 1.3 to 5.9% (mean = 3.5%). Meiospore infections in F(1)-generation larvae were significantly correlated with infections in parental-generation females, thus suggesting that larval infection rates could be substantially increased if methods were available to facilitate transmission of A. stimuli to a larger portion of the female population via inundative or inoculative release of infected copepods. No correlation was found when infections in filial-generation adult females were measured against meiospore infections in larvae from the preceding year. Analysis of yearly prevalence data using Fine's Fundamental Vertical Transmission Equation revealed low rates of horizontal transmission from the intermediate copepod host to female larvae in most years, ranging from 0.1 to 8.7% (mean = 3.1%). A. stimuli is enzootic, persists at a very low level, and has minimal impact on Ae. stimulans populations at this site. The low incidence rate of horizontal transmission to larvae appears to be due largely to a paucity of copepods and is a major factor that limits the abundance and subsequent proliferation of A. stimuli in Ae. stimulans populations at this locale. Results support the view that host-parasite cospeciation is an important mechanism of evolution in this group of mosquito/copepod microsporidia.     

Functional anatomy of the internal gills of the tadpole of Litoria ewingii (Anura,Hylidae)

Summary (1) Scanning electron microscopy and vascular casting were used to study the morphology and vascular anatomy of the fully developed internal gills of Litoria ewingii tadpoles. — (2) The four pairs of gills were located in two branchial baskets on either side of the heart. Each gill consisted of a branchial arch with gill tufts projecting ventrally and gill filters running dorsally. The gills bore a variable number of gill tufts in which a complex three-dimensional array of capillary loops, of varying lengths and diameters, was trailed in the path of the ventilatory current. — (3) The evidence presented in this paper suggests that the gill tufts have greater potential as gas exchangers than either the gill filters or skin. — (4) The study revealed structural and functional evidence for the existence of branchial shunts between afferent and efferent branchial arteries.     

Ecology of a diplozoon parasite on the gills of the African cyprinid Barbus neumayeri

In oxygen‐deficient waters, the difficulties of oxygen uptake in gill parasites and their fish hosts may influence host and parasite densities, site selection by the parasite, and effects of the parasite on host condition. This study quantified the prevalence and intensity of the gill monogenean Neodiplozoon polycotyleus in the African cyprinid fish Barbus neumayeri from an intermittent forest stream in western Uganda. Oxygen levels were low in the stream over the 12‐month study, averaging only 2.5 mg litre^?1 (monthly range = 1.2–4.3 mg litre^?1). However, parasite prevalence was high (47.2%), suggesting high tolerance to low oxygen in N. polycotyleus. The prevalence of parasites varied with host body size, with the highest frequency of occurrence in the middle size classes. Prevalence also varied over the year; seasonal peaks of rainfall coincided with a lower frequency of N. polycotyleus. The significantly nonrandom frequency distribution of parasites among hosts suggests regulation of parasite numbers. Of the hosts infected, 37.1% harboured one N. polycotyleus parasite, and 62.9% harboured two parasites. No fish were infected with more than two diplozoons. There was evidence for strong site specificity by N. polycotyleus within hosts; 77.7% of the parasites were located on the filaments of the second gill arch, which may relate to increased oxygen availability. In addition, only one of the 178 infected fish had more than one parasite on one side of the branchial basket. Although N. polycotyleus is undoubtedly parasitic, we found no evidence of a negative parasitic effect on the condition or reproductive status of B. neumayeri.     

Arteriovenous anastomoses in the afferent region of trout gill filaments (Salmo gairdneri richardson,teleostei)

Summary Arteriovenous anastomoses (AVAs) in the afferent region of trout gill filaments originate from two small filament arteries (Fromm's arteries), which parallel the main afferent filament vessel on either side. As in the efferent filament arteries the origin of AVAs is bordered by specialized endothelial cells. Fromm's arteries originate from efferent filament or branchial arteries. A few extremely narrow connections between the afferent filament artery and Fromm's arteries (= afferent shunts) do exist in some gill filaments. Nevertheless, the AVAs in the afferent filament region carry mainly arterialized blood, or blood plasma, to the central venous sinus of the filament.Supported by the Deutsche Forschungsgemeinschaft (Vo 229/1)     

Mast cell responses to Ergasilus (Copepoda), a gill ectoparasite of sea bream

Immunocytochemical, light microscopy and ultrastructural studies were conducted on gill of sea bream, Sparus aurata L., naturally parasitized with the important parasitic copepod Ergasilus sp. to assess pathology and cellular responses. Thirty-seven S. aurata were examined from a fish farm; 26 (70%) were parasitized, with infection intensity ranging from 3 to 55 parasites per fish. Hosts were divided into two groups, lightly infected fish (<15 parasites per fish) and heavily infected fish (>15 parasites per fish). In histological sections, the copepod encircled gill lamellae with its second antennae, compressed the epithelium, provoked hyperplasia and hemorrhage, occluded arteries and often caused lamellar disruption. Fusion of the secondary lamellae due to epithelial hyperplasia was common in all infected fish; heavily infected fish showed more intense branchial inflammation. In both healthy and infected fish, mast cells (MCs) were free within the connective tissue inside and outside the blood vessels of the primary lamellae and made close contact with vascular endothelial cells, mucous cells and rodlet cells (RCs). MCs were irregular in shape with a cytoplasm filled by numerous electron-dense, membrane-bound granules. Immunostaining of primary and secondary gill filaments with an antibody against the antimicrobial peptide (AMP) piscidin 3 (anti-piscidin 3 antibody, anti-HAGR) revealed a subpopulation of MCs that were positive. These MCs were more abundant in gills of heavily infected fish than in either lightly infected or uninfected fish (ANOVA, P<0.05). Our report documents the response of gill to ectoparasite infection and provides further evidence that mast cells and their AMPs may play a role in responding to branchial ectoparasite infections.     

Potential negative effects and heterogeneous distribution of a parasitic copepod Salmincola edwardsii (Copepoda: Lernaeopodidae) on Southern Asian Dolly Varden Salvelinus curilus in Hokkaido,Japan

The genus Salmincola is an ectoparasitic copepod group commonly infesting the branchial and buccal cavities of salmonids. While negative impacts on hatchery fishes have been reported, their impacts on wild fish populations and distribution patterns are critically understudied. In the Shiretoko Peninsula, Hokkaido, Japan, we found parasites belonging to this genus on the branchial cavity of a stream salmonid, Southern Asian Dolly Varden Salvelinus curilus. All parasites recovered were identified as Salmincola edwardsii based on morphological characteristics and partial 28S rDNA sequences. Prevalence was highly heterogeneous even among neighboring streams (0–54.8%, < 10 km) with the mean intensity among streams being generally low (2.19 parasites/infeted fish). Despite the low intensity, quantile regression analysis showed negative trends between parasite intensity and host condition, suggesting that the infestation of S. edwardsii has a potential negative impact on the host salmonid. In addition, a single copepod was found from an anadromous fish, which could indicate some salinity tolerance of the copepods. It is important to evaluate the effects of Salmincola spp. on host species and determine the limiting factors on the parasite's distribution for proper management.     

Oysters as habitat for limpets in a temperate mangrove forest

In temperate mangrove forests in New South Wales, Australia, the limpet Patelloida mimula Iredale lives on the oyster Saccostrea commercialis Iredale and Roughley, which, along with mangrove trees, provide the only hard substrata in a habitat otherwise dominated by soft-sediment. The objective of this study was to ascertain the degree of association between the limpet and the oyster by examining their patterns of co-occurrence in the forest and the relationship between individual pairs of limpets and oysters. Sampling of the distribution and abundance of limpets and oysters throughout the mangrove forest revealed that limpets were rarely present on substrata other than oysters. Patterns of abundance of limpets were, however, not directly related to the amount of habitat provided by the oysters. For example, there was a dramatic decline in the abundance of oysters from seaward to landward in the mangrove forest which was not reflected in the densities of limpets. Consequently, oysters appear to set the limits of distribution of limpets, but other factors modify their broad-scale patterns of distribution and abundance within these bounds. Limpets leave scars on oysters which might be home sites. About 98% of oysters with limpets had only one limpet per surface of oyster, and the distribution of limpets was overdispersed or repulsed. Moreover, in some areas of the forest, the lengths of limpets were directly related to lengths of oysters. Tracking of individual limpets for 13 days revealed that > 70% remained on the same scar of an oyster. This evidence suggests that for some limpets there is a strong association with particular oysters. Factors determining the distribution, abundance, and sizes of oysters are likely to be important sources of variation to the structure of populations of P. mimula in temperate mangrove forests.     

Functional Organization of the Teleost Gill

The circulation of the gills has been studied in the perch, trout and eel combining the conventional histological methods and casting techniques. The existence of two blood pathways in each gill arch was confirmed. 1 — An arterio-arterial pathway assuming the respiratory function. It includes the afferent branchial artery and in each primary lamella the afferent primary artery, the secondary lamellae capillaries and the primary and branchial efferent arteries. 2 — An arterio-venous pathway arising from both the branchial artery, in the gill arch, and the primary arteries in each primary lamella. This pathway includes the central venous sinus of the primary lamella, several small veins and is finally connected with the branchial veins. 3 — The lack of connections between afferent primary arteries and cvs in the trout and the perch makes impossible a direct blood flow from the afferent to the efferent artery (shunt). In the eel connections between cvs and both afferent and efferent arteries do not mean that a shunt is operating according to the pressure gradient.     

Some ecological aspects of the copepod parasites of the common hake, Merluccius hubbsi, from the Argentine-Uruguayan coasts

A total of 2164 common hake Merluccius hubbsi captured in the Argentine-Uruguayan Common Fishing Zone was examined for parasitic copepods. The fish were infested with Chondracanlhus palpifer and Neobrachietia insidiosa f. lageniformis, the former parasitizing the buccal cavity and the latter the branchial arches. C. palpifer showed an increase in both prevalence and intensity in relation to the host size; infestation with N. insidiosa f. lageniformis decreased with increasing length of hake. Attachment site preferences and variations in the parasite distribution patterns as related to the host biology were observed. Evidence of negative association between copepod species and seasonal changes in the parasite composition was also found.     

Copepod ectoparasites of Merlangius merlangus and Platichthys flesus

Seasonal changes in the levels of copepod ectoparasitic infections from Merlangius merlangus and Platichthys flesus were examined and related to the annual migrations of young fish into an estuary. Lernaeocera branchialis (L.) was the only parasite common to both fish species; its life-cycle stages infected the branchial chambers of 96·7% flounders ( P. flesus ) and 40·4% whiting ( M. merlangus ). Infection in the two fish species exhibited similar seasonal fluctuations and a temporal relationship between the parasite's occurrence on its two hosts was apparent. Whiting were also parasitized by Clavella uncinata (Milller); 26·9 % fish were affected. The copepods' distribution was highly localized with the majority restricted to the long gill rakers of the first gill arch. Infection was related to size of whiting up to 12·5 cm; there was little increase beyond this length. In addition to L. branchialis , flounder was also host to Acanthochondria depressa (T. Scott) and Lepeophtheirus pectoralis (Müller). A. depressa showed a preference for the ocular side of the body with its developing stages attached mainly to the second gill arch and the adults located in the posterior pocket of the branchial chamber. L. pectoralis was associated with the paired fins. Factors which could result in such localized distributions are discussed.     

Parasite fauna of introduced pumpkinseed fish Lepomis gibbosus: first British record of Onchocleidus dispar (Monogenea)

A survey of pumpkinseed Lepomis gibbosus from a stillwater fishery in southern England revealed infections by the ancyrocephalid monogenean Onchocleidus dispar at 100% prevalence. Host specificity of O. dispar to North American centrarchid fish suggests that it is a non-native parasite, introduced to Britain with L. gibbosus. Mean intensity of O. dispar was significantly higher in male (25.5 parasites) compared to female (15) and immature (7) hosts, but was not influenced by host standard length or sampling time. This sex-biased parasitism is likely to be related to both ecological and physiological factors, such as differential exposure to infective stages during nest building, or higher susceptibility to infection due to enhanced host stress levels. O. dispar significantly dominated the external surfaces and the outer gill arches. Analysis of the spatial distribution of the parasite between the 9 regions of gill surface determined that as density increased, higher numbers of parasites were found on the anterior and central gill regions compared to posterior (distal) regions. Apparent microhabitat selection is discussed in terms O. dispar life cycle, maximising oxygen availability, avoiding unstable areas and increasing intraspecific contact. The absence of O. dispar in the gills of roach, rudd and gudgeon sampled from the same fishery supports the assumption that this parasite is currently of little threat to native fish populations.     

Die Vaskularisation der Kiemen von Myxine glutinosa L. (Cyclostomata)

The angioarchitecture of the gills in Myxine glutinosa L. was studied by scanning electron microscopy of vascular corrosion casts (methylmethacrylate). It was found that the afferent branchial artery may be connected to the sinus peribranchialis by a papilla. The sinus is connected to delicate vessels and sinuses, which are interposed between meridionally arranged radial arteries. These delicate vascular formations continue into the interior of the gill and form a plexus on both poles of the gill folds. Light and scanning electron microscopical studies on the vascular endothelium of the afferent vessels of the gill lamellae reveal rounded endothelial cells, which are characterized by a high content of granula, by long cellular processes and by bridge-formations towards neighbouring cells. Supporting columns within the vascular system of the lamellae were found to be set up by several spirally arranged cells. SEM observations reveal goblet cells and numerous superficial epithelial cells with various numbers of microvilli and microridges forming the epithelial surface of the gill folds.     

Microhabitats of gill parasites (Monogenea and copepoda) of teleosts (Scomberoides spp.)

Ramasamy P., Ramalingam K., Hanna R. E. B. and Halton D. W. 1985. Microhabitat of gill parasites (Monogenea and Copepoda) of teleosts (Scomberoides spp.). International Journal for Parasitology15: 385–397. The parasites Vallisia indica, Allodiscocotyla chorinemi, Heterapta chorinemi, and Dionchus remorae exhibited site specificity on the gills of Scomberoides commersonianus, S. tol, S. lysan and S. tala, whereas the copepod Caligus sp. did not. Observations on site preference revealed microhabitat differences along the length of gill filaments, along the anterio-posterior axis of the gill, between external and internal gill filaments and on different gill arches. Site preference varied with parasite density on each gill and host. An interspecific association test between pairs of species revealed, in some cases, a positive association, and in other cases a negative association. There were apparently no association? between certain pairs of species. A comparison of intensity within pairs of parasite species infecting the same host revealed either an inverse or a direct correlation. The numerical dominance and prevalence of parasites differed on each host species. This study indicates that intra- and interspecific competition may occur among the gill parasites. The direction and speed of ventilation water-currents and certain intrinsic factors of the parasite themselves may determine their microhabitat restriction on the gills.     

An alternative model to study the association of rainbow trout (Oncorhynchus mykiss L.) pathogens with the gill tissue

This study describes the development of a trout gill perfusion model, consisting of an excised branchial arch from rainbow trout (Oncorhynchus mykiss L.), perfused via the afferent branchial artery and suspended in a circular organ chamber filled with Ringer solution. Different perfusion fluids were tested: Ringer, Cortland, Ringer + procaine, Ringer + adrenalin, Cortland + procaine, Cortland + adrenalin and Cortland + dextran 1%. The latter perfusion fluid proved to be satisfactory, maintaining the gill tissue in a healthy condition outside the body of the fish for at least 180 min. Using this model, the interaction of damaging agents with the trout gill tissue may be studied under carefully controlled conditions. The trout gill perfusion model leads effectively to a reduction in the number of experimental animals to be used and also involves an elimination of pain and/or suffering, which is as good as complete.