Alexandre Lobo-da-Cunha

University of Porto, Oporto, Porto, Portugal

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Publications (36)76.18 Total impact

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    ABSTRACT: Twenty two strains of Rhodopirellula were isolated from the epiphytic community of several marine macroalgae and separated into two groups, designated as group B and group C. In this study, we characterized these groups as two novel species belonging to the genus Rhodopirellula. These strains were represented by pleomorphic cells that were arranged in rosettes and formed pink- or red-pigmented colonies. The organisms were chemoorganotrophic and required vitamin B12 for growth. Their optimal temperature for growth was around 25 °C. Major fatty acids were C18:1 ω9c, C16:0 and C16:1 ω7c/C16:1 ω6c. Phosphatidylcholine and phosphatidylglycerol were the major polar lipids. Unidentified phospholipids were also present. The 16S rDNA sequence analysis confirmed the affiliation of these organisms to the order Planctomycetales, genus Rhodopirellula, with R. baltica as the closest phylogenetic relative. The analysis of a partial sequence of the gene encoding the β-subunit of RNA polymerase (rpoB) confirmed the phylogenetic separation of the isolates into two different species of the genus Rhodopirellula. The 16S rRNA sequences from strains of group B revealed their widespread occurrence across the world, whereas strains of group C were not observed before. On the basis of physiological, biochemical, chemotaxonomic and genetic characteristics we propose that our isolates represent two new species of Rhodopirellula, Rhodopirellula rubra sp. nov. (type strain is LF2T = DSM 25,459 = CECT 8075) and Rhodopirellula lusitana sp. nov. (type strain is UC17T = DSM 25,457 = LMG 27,777).
    Systematic and Applied Microbiology 01/2014; · 3.29 Impact Factor
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    Olga Maria Lage, Joana Bondoso, Alexandre Lobo-da-Cunha
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    ABSTRACT: Knowledge of the interesting phylum of Planctomycetes has increased in the last decades both due to cultural and molecular methods. Although a restricted number of species have been described to date, this group presents a much larger diversity that has been mainly revealed by molecular ecology studies. Isolation experiments allowed us to get a number of new Planctomycetes taxa that extend the already described ones. In this work we present the ultrastructural morphological characterization of these new taxa as well as we give new details of Aquisphaera giovannonii ultrastructure. Furthermore, our interpretation on Planctomycetes cell envelope is provided.
    Antonie van Leeuwenhoek 07/2013; · 2.07 Impact Factor
  • Catarina Inês Gonçalves, Alexandre Lobo-da-cunha
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    ABSTRACT: This study was conceived to evaluate seasonal changes in the lipid reserves of the digestive gland of the carnivorous intertidal caenogastropod Nucella lapillus, using a stereological approach. Volume density of lipid droplets in the digestive gland, digestive gland weight and volume were assessed in animals collected in March, June, September and December on the Portuguese coast. Gonad development was evaluated to detect any relationship between lipid content in the digestive gland and the reproductive cycle. The quantitative light microscopic analysis demonstrates that lipid droplets are a major component of the digestive gland. In males, the digestive gland and its lipid reserves were quite stable without significant variations throughout the year. In females, the percentage of digestive gland volume occupied by lipid droplets was higher in June and December, coinciding with the highest values of digestive gland volume. Due to the conjugation of these two factors, in June and December the total amount of lipids in the digestive gland was substantially increased in females. In both males and females a relationship between the development status of the gonad and the lipid reserves of the digestive gland was not evident. However, significant differences in the digestive gland lipid reserves were detected between males and females in June and December, pointing to a sex related effect on lipid reserves. To evaluate the use of lipid reserves as energy source in N. lapillus, the consumption of digestive gland lipids was followed during a starvation experiment.
    Journal of the Marine Biological Association of the UK 05/2013; 93(03). · 1.02 Impact Factor
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    ABSTRACT: In this study, semi-thin sections stained with histochemical techniques and transmission electron microscopy were used to obtain new data about the morphology and function of the male copulatory apparatus of the cephalaspidean gastropod Bulla striata. The apparatus comprises a vestibule, a penial papilla and a prostate consisting of a coiled unbranched tube ending in a blind caecum. The penial papilla and the coiled tubular prostate are enclosed by a muscular sheath, which is continuous with the muscular tissue of the vestibule. The epithelium lining the lumen of the vestibule is formed by ciliated and mucus-secreting cells. Two new types of subepithelial secretory cells were discovered in this region. The penial papilla is a muscular structure without secretory cells in the epithelium lining the narrow lumen. The tubule that constitutes the prostate possesses a muscular wall and can be divided in three distinct regions: a non-secretory duct connected to the penial papilla, a glandular region rich in large secretory cells and the terminal caecum containing just a few small secretory cells. In the terminal blind caecum, the muscular sheath is fused with the muscular wall of the tubular prostate. Large numbers of spermatozoa were found in the glandular region and in the terminal caecum of the prostate. A new functional mechanism is proposed to explain penial eversion during copulation. This differs from a previous hypothesis in two main aspects: (1) existence of a permanent penial papilla in mature animals acting as a functional penis and (2) functional role of vestibule during copulation, which everts and surrounds the penial papilla, while the latter protrudes outwards.
    Zoomorphology 01/2013; 132(2):145. · 1.13 Impact Factor
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    ABSTRACT: A negative correlation between female gonadal maturation kinetics and size variations of hepatic peroxisomes was earlier documented in brown trout, as a probable impact of serum estrogen changes during the reproductive cycle. Herein, we investigated whether the organelle volume/surface dynamics seen in female brown trout liver peroxisomes - without numerical changes within each hepatocyte - is followed by variations in the expression of the membrane peroxisome protein Pex11α gene. For comparison, we also studied males. We find in females a seasonal variation with the highest Pex11α expression in February, which was statistically different from all other tested periods. Overall, the expression of PEX11α had over a fivefold decrease from February to September. This period coincides with the reproductive transition between the earlier post-spawning gonadal remodeling and preparatory staging and the pre-spawning period. Males did not show changes. Our approach allowed the first characterization of a peroxin gene in a teleost, the Pex11α, while offering a correlation scenario were, as we hypothesized, the peroxisomal size kinetics is paralleled by membrane-related gene alterations (measured herein as proxy of Pex11α gene expression). Our data support and expand previous results on the regulation, function and morphology of peroxisome dynamics in brown trout, with a broader interest.
    Comparative biochemistry and physiology. Part A, Molecular & integrative physiology 09/2012; · 2.20 Impact Factor
  • A. D. Resende, A. Lobo-da-Cunha, E. Rocha
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    ABSTRACT: Fish renal morphology has been studied since the 1960’s, but is still incomplete for most species, even for some of the most studied. The functional and morphological diversity and specialization of the fish kidney, due to environmental adaptations, makes this a large subject that remains understudied. References to studies on fish renal structure are given in Table 1. Although authors as Hentschel and Elger contributed to the knowledge of fish renal morphology in a comparative perspective, the knowledge concerning this issue is still reduced. Here we give a short comparative summary of fish renal morphology, using as baseline model the brown trout.
    Microscopy and Microanalysis 08/2012; 18(S5):47-48. · 2.50 Impact Factor
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    ABSTRACT: A Gram-positive, aerobic, non-motile, endospore-forming rod, designated DS22(T), was isolated from a drinking-water treatment plant. Cells were catalase- and oxidase-positive. Growth occurred at 15-37 °C, at pH 7-10 and with <8% (w/v) NaCl (optimum growth: 30 °C, pH 7-8 and 1-3% NaCl). The major respiratory quinone was menaquinone 7, the G+C content of the genomic DNA was 36.5 mol% and the cell wall contained meso-diaminopimelic acid. On the basis of 16S rRNA gene sequence analysis, strain DS22(T) was a member of the genus Bacillus. Its closest phylogenetic neighbours were Bacillus horneckiae NRRL B-59162(T) (98.5% 16S rRNA gene sequence similarity), Bacillus oceanisediminis H2(T) (97.9%), Bacillus infantis SMC 4352-1(T) (97.4%), Bacillus firmus IAM 12464(T) (96.8%) and Bacillus muralis LMG 20238(T) (96.8%). DNA-DNA hybridization, and biochemical and physiological characterization allowed the differentiation of strain DS22(T) from its closest phylogenetic neighbours. The data supports the proposal of a novel species, Bacillus purgationiresistans sp. nov.; the type strain is DS22(T) (=DSM 23494(T)=NRRL B-59432(T)=LMG 25783(T)).
    INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY 02/2011; 62(Pt 1):71-7. · 2.11 Impact Factor
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    ABSTRACT: As part of a study of the diversity of planctomycetes, two novel strains, designated OJF2(T) and OJF8, were isolated from the sediments of a freshwater aquarium. The organisms were chemoheterotrophic, spherical and pink-pigmented, had an optimum growth temperature of about 30-35 °C and an optimum pH for growth of around 7.5-8.5. The predominant fatty acids were C(18:1)ω9c and C(16:0). The two strains were able to assimilate several sugars and organic acids. 16S rRNA gene sequence analysis confirmed the affiliation of these organisms to the phylum 'Planctomycetes'; they showed highest similarity to the type strains of Singulisphaera acidiphila (92.4%) and Isosphaera pallida (91.9%). On the basis of physiological, biochemical and chemotaxonomic characteristics, strains OJF2(T) and OJF8 are considered to represent a novel species of a new genus of the order Planctomycetales, for which the name Aquisphaera giovannonii gen. nov., sp. nov. is proposed. The type strain of Aquisphaera giovannonii is OJF2(T) (=CECT 7510(T) =DSM 22561(T)).
    INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY 01/2011; 61(Pt 12):2844-50. · 2.11 Impact Factor
  • Journal of Molluscan Studies 01/2011; 77(4):322-331. · 1.36 Impact Factor
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    ABSTRACT: The large variability in kidney morphology among fish makes it difficult to build a "universal" model on its function and structure. Therefore, a morphological study of brown trout trunk kidney was performed, considering potential seasonal and sex effects. Three-year-old specimens of both sexes were collected at four stages of their reproductive cycle. Kidney was processed for light and electron microscopy. The relative volumes of renal components, such as renal corpuscles and different nephron tubules, were estimated by stereological methods. Qualitatively, the general nephron structure of brown trout was similar to that described for other glomerular teleost species. Quantitatively, however, differences in the relative volume of some renal components were detected between sexes and among seasons. Particularly, highest values of vacuolized tubules and new growing tubules were observed after spawning, being more relevant in females. Despite seasonal changes, more linear correlations were found between those parameters and the reno-somatic index than the gonado-somatic index. Thus, we verified that some brown trout renal components undergo sex dependent seasonal variations, suggesting a morphological adaptation of the components to accomplish physiological needs. These findings constitute a baseline for launching studies to know which factors govern the morphological variations and their functional consequences.
    Microscopy and Microanalysis 10/2010; 16(6):677-87. · 2.50 Impact Factor
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    ABSTRACT: The posterior esophagus of Bulla striata, running from the gizzard to the stomach, was investigated with light and electron microscopy to obtain new data for a comparative analysis of the digestive system in cephalaspidean opisthobranchs. In this species, the posterior esophagus can be divided into two regions. In the first, the epithelium is formed by columnar cells with apical microvilli embedded in a cuticle. Many epithelial and subepithelial secretory cells are present in this region. In both, electron-lucent secretory vesicles containing filaments and a peripheral round mass of secretory material fill the cytoplasm. These acid mucus-secreting cells may also contain a few dense secretory vesicles. In the second part of the posterior esophagus, the cuticle is absent and the epithelium is ciliated. In this region, epithelial cells may contain larger lipid droplets and glycogen reserves. Subepithelial secretory cells are not present, and in epithelial secretory cells the number of dense vesicles increases, but most secretory cells still contain some electron-lucent vesicles. These cells secrete a mixture of proteins and acid polysaccharides and should be considered seromucous. The secretory cells of the posterior esophagus are significantly different from those previously reported in the anterior esophagus of this herbivorous species.
    Microscopy and Microanalysis 10/2010; 16(6):688-98. · 2.50 Impact Factor
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    ABSTRACT: Sex differences exist in fish hepatocytes, but studies for characterizing their cytology throughout the breeding cycle are still scarce; suggesting changes, but most lacking quantitative data. To address this limitation, to complement baseline data generated from the brown trout model, and to prove that sex-specific seasonal changes exist, we made an unbiased stereological evaluation of the hepatocytic cytoplasm. Unprecedentedly for fish liver, the stereological design was exempt from model (biased) assumptions. Five (3 years old) animals per sex were studied in endogenous vitellogenesis, exogenous vitellogenesis, and spawning season end. Liver pieces for analysis were systematically sampled. Stereology was done in transmission electron microscopy (TEM) micrographs. Primary data generated relative volume estimates of the major cytoplasmic components. Such values were used for deriving absolute volumes (per cell and per liver). Lipid droplets did not show changes. As to other targets, trends at cell and liver levels were not always equal. If the hepatocyte was the reference space, the contents in mitochondria, dense bodies, glycogen, and cytosol changed seasonally, in both sexes. If taking the liver as the reference, changes attained the Golgi apparatus and rough endoplasmic reticulum (RER), besides dense bodies, glycogen (in females), and cytosol. The components volumes (namely per liver) were often positively (negatively for glycogen) correlated with the ovary weight, disclosing new associations and implications in fish. While also offering gold-standard data for backing morphofunctional correlations and pathology, we revealed a new process by which females increase the amount of RER and Golgi throughout vitellogenesis, breaking from the idea on how this event happens in fish.
    Microscopy Research and Technique 08/2010; 73(8):766-78. · 1.59 Impact Factor
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    ABSTRACT: Neurotoxicity induced in fish by domoic acid (DA) was assessed with respect to occurrence of neurotoxic signs, lethality, and histopathology by light microscopy. Sparus aurata were exposed to a single dose of DA by intraperitoneal (i.p.) injection of 0, 0.45, 0.9, and 9.0 mg DA kg(-1) bw. Mortality (66.67 ± 16.67%) was only observed in dose of 9.0 mg kg(-1) bw. Signs of neurological toxicity were detected for the doses of 0.9 and 9.0 mg DA kg(-1) bw. Furthermore, the mean concentrations (±SD) of DA detected by HPLC-UV in extracts of brain after exposure to 9.0 mg DA kg(-1) bw were 0.61 ± 0.01, 0.96 ± 0.00, and 0.36 ± 0.01 mg DA kg(-1) tissue at 1, 2, and 4 hours. The lack of major permanent brain damage in S. aurata, and reversibility of neurotoxic signs, suggest that lower susceptibility to DA or neuronal recovery occurs in affected individuals.
    Marine Drugs 01/2010; 8(10):2721-32. · 3.98 Impact Factor
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    Alexandre Lobo-da-Cunha, Iris Ferreira, Rita Coelho, Gonçalo Calado
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    ABSTRACT: Cephalaspideans are a group of opisthobranch gastropods that comprises carnivorous and herbivorous species, allowing an investigation of the relationship between these diets and the morphofunctional features of the salivary glands. In this study, the salivary glands of the carnivorous cephalaspidean Philinopsis depicta were observed by light and electron microscopy. The secretory epithelium of these ribbon-shaped glands is formed by ciliated cells, granular cells and cells with apical vacuole. In ciliated cells the nucleus and most cytoplasmic organelles are located in the wider apical region and a very thin stalk reaches the base of the epithelium. These cells possess significant amounts of glycogen. Granular cells are packed with electron-dense secretory granules and also contain several cisternae of rough endoplasmic reticulum and Golgi stacks. The other type of secretory cell is mainly characterized by the presence of a large apical vacuole containing secretion. These cells possess high amounts of rough endoplasmic reticulum cisternae and several Golgi stacks. Vesicles with peripheral electron-dense material are also abundant, and seem to fuse to form the apical vacuole. The available data point out to a significant difference between the salivary glands of carnivorous and herbivorous cephalaspidean opisthobranchs, with an intensification of protein secretion in carnivorous species.
    Tissue and Cell 05/2009; 41(5):367-75. · 1.04 Impact Factor
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    ABSTRACT: Previously, it was documented that liver peroxisomes display seasonal size changes in the adult Salmo trutta fario, especially in females (and negatively correlated with ovary maturation). It was then hypothesized that decreases in peroxisome size could be paralleled by changes in peroxisomal beta-oxidation and estradiol catabolism actions. The 17beta-hydroxysteroid dehydrogenase 4 has been portrayed as playing an important role in both processes. To elucidate its function in the described peroxisomal pattern, we isolated the cDNA and predicted the protein sequence of the enzyme in that species. The seasonal gene expression pattern in both genders was addressed through quantitative PCR. Fish sampling was in post-spawning period, early and advanced gonad maturation, and pre-spawning. Males did not vary seasonally. As to females, a seasonal pattern was evidenced according to our previous hypothesis. We suggest that the decreased levels observed during vitellogenesis are related to lipid needs for ovary maturation, and, additionally, with the need of modulating estradiol titers.
    Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology 04/2009; 153(2):157-64. · 1.61 Impact Factor
  • Eniko Kadar, Alexandre Lobo-da-Cunha, Carlos Azevedo
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    ABSTRACT: Calcium carbonate transfer was experimentally examined in hydrothermal mussels Bathymodiolus azoricus, which were collected from 850m depth at Menez Gwen hydrothermal vent site (31°31′W, 37°50′N) on the Mid Atlantic Ridge in May 2007. In each of four 10-day experiments, groups of mussels were maintained at atmospheric pressure or re-pressurised to depths relevant to their site of occurrence, i.e. 850m depth at Menez Gwen, 1,750m at Lucky Strike (31°31′W, 37°18′N) and 2,300m at Rainbow (31°31′W, 36°13′N). The shells of experimental mussels were perforated and mantle tissue was fixed for light and TEM studies at days 7 and 10 following the injury. Simultaneously, haemocytes from the extrapallial fluid (EPF) at the site of induction were studied. At day 7 the response was most intense in the middle fold of the mantle margin and possibly proportional to hydrostatic pressure. At day 10 the epithelial cells on the mantle surface facing the body cavity produced copious organic secretions that avidly bound calcium. Haemocyte migration was noted within the mantle tissue, and the haemocytes at the mantle surface facing the shell had a Ca-positive granular content. Large haemocytes were detected in the EPF at the injury site, and some showed evidence of an immune reaction while others showed Ca-positive granular content. These results suggest that haemocytes are involved in shell repair in these deep-sea mussels just as in some freshwater and shallow marine molluscs.
    Marine Biology 03/2009; 156(5):959-967. · 2.47 Impact Factor
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    ABSTRACT: Lobo-da-Cunha, A., Oliveira, E., Alves, Â., Coelho, R. and Calado, G. 2010. Light and electron microscopic study of the anterior oesophagus of Bulla striata (Mollusca, Opisthobranchia). —Acta Zoologica (Stockholm) 91: 125–138.The anterior oesophagus of Bulla striata was investigated with light and electron microscopy. In the most anterior region, the ridges of the oesophageal wall are covered by a ciliated columnar epithelium forming large apical blebs which are released into the lumen, an activity that is particularly intense in the oesophageal pouch. In the last two-thirds of the anterior oesophagus, the epithelium is covered with microvilli embedded in a cuticle, but apocrine secretion and cilia are absent. Subepithelial secretory cells are very abundant in the oesophageal wall, except in the roof of the pouch. They have a long neck that crosses the epithelium, whereas the cell body containing the nucleus is embedded in the connective tissue. Large electron-lucent secretory vesicles and many Golgi stacks fill most of their cytoplasm. The histochemical and cytochemical assays show that these cells secrete acid mucopolysaccharides. With the current and future studies we aim to obtain data for the establishment of relationships between morphofunctional features of the digestive system and food types in cephalaspideans. Additionally, the new data about the oesophageal pouch of B. striata may be useful for the establishment of eventual homologies with the oesophageal diverticula of other opisthobranchs.
    Acta Zoologica 02/2009; 91(2):125 - 138. · 1.47 Impact Factor
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    ABSTRACT: PPAR isotypes have been previously identified in the teleost brown trout (Salmo trutta f. fario) and their organ distribution pattern established. Being that the liver is a vital metabolic organ presenting expression of all isotypes and also knowing that estrogens/estrogen receptors seem to interact with PPARs, we hypothesized that the latter may very well change seasonally. So, we studied the expression of these receptors in the liver, along the annual reproductive cycle and in both genders. According to real-time RT-PCR, PPARalpha mRNA expression in females was significantly higher in May and lower in September than in other seasons. No significant variation was observed along the year in males. A significant difference between genders occurred in May, when PPARalpha expression was higher for females. PPARbeta expression showed little variation along the reproductive cycle in females, but in males it was significantly higher in December than in the other seasons. No significant differences existed between genders. PPARgamma was more expressed in February than in September and December, for females. As to males, it was more expressed in February than in all other seasons. No significant differences were observed between genders. The study proved our hypothesis that PPARs gene expression varies along the year. Moreover, PPARalpha expression in females followed the same annual variation pattern as peroxisome volumes and enzyme activities, and an inverse pattern relatively to the salmonid type annual plasma estradiol levels. The data agrees with the idea that PPARalpha is under estradiol modulation and that cross-talk between this receptor and the estrogen receptor possibly exists.
    General and Comparative Endocrinology 01/2009; 161(1):146-52. · 2.82 Impact Factor
  • A. Lobo-da-Cunha, I. Ferreira, G. Calado
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    ABSTRACT: Cephalaspideans are a group of opisthobranch gastropods comprising carnivorous and herbivorous species, allowing an investigation of the relationship between these diets and the morphofunctional features of the salivary glands.In this study, the salivary glands of the carnivorous cephalaspidean Philinopsis depicta were observed by light microscopy using semithin sections and by transmission electron microscopy. A central duct runs along the length of these thin ribbon-shaped glands dividing them in two halves, each formed by a single row of tubules perpendicularly attached to the central duct. The simple epithelium of the central duct and lateral tubes contains ciliated cells and two types of secretory cells, named granular cells and cells with apical vacuole (Fig. 1). A very thin outer layer of connective tissue covers the epithelium (Fig. 1). The ciliated cells are numerous but very thin, forming small clusters between secretory cells. The nucleus, several mitochondria and a few lysosomes are located in the apical region were the cells are wider. A very thin cytoplasmic stalk reaches the base of the epithelium and contains bundles of filaments in addition to some mitochondria.
    Microscopy and Microanalysis 01/2009; 15. · 2.50 Impact Factor
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    Enikö Kádár, Sean A. Davis, Alexandre Lobo-da-Cunha
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    ABSTRACT: Invertebrates harbouring endosymbiotic chemoautotrophic bacteria are widely distributed in a variety of reducing marine habitats, including deep-sea hydrothermal vents. Bathymodiolids are dominants of the biomass at geochemically distinct vent sites of the Mid Atlantic Ridge (MAR) and thus are good candidates to study biological processes in response to site-specific conditions. To satisfy their nutritional requirements, these organisms depend to varying extent on two types of chemoautotrophic symbionts and on filterfeeding. The quantitative relationships of the nutritional modes are poorly understood. Using enzyme cytochemistry, electron microscopy and X-ray microanalysis, the structural and functional aspects of the cellular equipment necessary for lysosomal digestion was studied. We provide evidence for the following: (1) the basis of intracellular digestion of symbionts in Bathymodiolus azoricus from two geochemically distinct vent sites was not mainly in the large lysosomal bodies as previously thought (based on the membranous content resembling bacteria); (2) senescent bacteria are autolysed, possibly by bacterial acid phosphatase, that is more likely a cell cycling of the symbionts rather than an active lysosomal digestion by the host; (3) the consistent absence of hydrolases may indicate the improper use of the name “lysosome” for large vesicles at the base of the gill bacteriocytes (4) nutrient transfer in B. azoricus, therefore, may more likely be accomplished through leaking of metabolites from the symbiont to the host, not excluding lysosomal resorption of dead bacteria as an auxiliary strategy for organic molecule transfer; (5) evidence is provided for microvillar transfer of substances from the seawater that may indicate filter-feeding, in non-symbiotic ciliated gill cells of mussels from Lucky Strike; (6) two types of lysosomal vesicles can be distinguished in digestive cells based on their enzymatic content and their elemental composition.
    Marine Biology 01/2008; 153(5):995-1004. · 2.47 Impact Factor