Spawning of the colonial coral <Emphasis Type="Italic">Cladocora caespitosa</Emphasis> (Anthozoa,Scleractinia) in the Southern Adriatic Sea

Data on sexual reproduction of scleractinian coral species living in temperate zones, particularly in the Mediterranean Sea, are quite scarce. This paper describes sexual reproduction of the colonial coral Cladocora caespitosa from Veliko jezero (Mljet Island) in the Adriatic Sea. Spawned orange eggs and white sperm bundles were observed on the coral bank of C. caespitosa two nights before the full moon (20 June 2005) coinciding with increasing water temperature and correlated with the lunar cycle. Spawning was observed during five nights, involving about 30% of the colonies from the coral bank. Different colonies on the bank released only one type of gamete during the reproductive period. The diameter of the sperm bundles ranged from 100 to 200 μm (average 163 μm; SD = 47.08), while the female gametes diameter ranged from 300 to 500 μm (average 416 μm; SD = 73.12).     

The impact of seawater temperature on coral growth parameters of the colonial coral Cladocora caespitosa (Anthozoa, Scleractinia) in the eastern Adriatic Sea

The scleractinian coral Cladocora caespitosa deserves a special place among the major carbonate bioconstructors of the Mediterranean Sea. Annual coral skeleton growth, coral calcification, and skeleton density of the colonial coral C. caespitosa taken from 25 locations in the eastern Adriatic Sea were analyzed and compared with annual sea surface temperatures (SST). The growth rates of the coral C. caespitosa from the 25 stations in the Adriatic Sea ranged from 1.92 to 4.19?mm per year, with higher growth rates of the investigated corallites in the southern part of the Adriatic Sea. These growth rates are similar to those measured in other areas of the Mediterranean Sea. The correlation between coral growth and sea temperatures in the Adriatic Sea is seen as follows: An X-radiograph analysis of coral growth in C. caespitosa colonies that are over 60?years old showed that higher growth rates of this coral coincided with a warmer period in the Mediterranean Sea. A positive significant correlation exists between corallite growth rates and SST and coral calcification and SST. A negative correlation exists between coral density and SST. Coral growth rates also showed a correlation with higher eutrophication caused by nearby fish farms, along with a greater depth of the investigated colonies and high bottom currents.     

Release of sperm clusters in spheres by the black coral <Emphasis Type="Italic">Cupressopathes pumila</Emphasis> (Anthozoa,Antipatharia)

Fecund polyps from a bottlebrush-shaped colony of black coral Cupressopathes pumila (Brook 1889) from Ambon Island (Maluku Archipelago, Indonesia) were studied at the structural and ultrastructural levels. Five fragments, each 5 cm long and containing about 60 polyps, were cut from a single parental colony. The fragments underwent different treatments: two were fixed in 70% alcohol for species identification; the remaining three were reared in aerated aquaria and then processed for anatomic studies by light and transmission electron microscopy. Some polyps from one of these reared fragments displayed spheres of various sizes protruding from the mouth. Four hours after sampling, spheres settled on the bottom of the aquarium. Comparative analysis of the fecund polyps showed: (i) the differentiation of spermatocysts inside the transverse primary mesenteries that separate the lateral tentacles from one another; (ii) spermatocysts entering the gastric cavity after breakage of the mesentery wall and (iii) location of spermatocysts close to the basal part of the pharynx. Analysis of the largest spheres, both close to the external surface of the mouth and settled on the bottom of the small aerated aquaria, showed that they were irregularly bordered with residual mesentery tissue and contained clusters of spermatocysts. Gametes presented various phases of differentiation up to the final sperm, as it occurs in the spermatocysts developing inside the mesenteries of the fecund polyps. In consideration of the particular distribution of the colonies of C. pumila, which grow very far apart, the release of buoyant spheres is a particular modality of spawning which can be viewed as a strategy for successful fertilisation over long distances.     

Intraspecific contact and egg production in the massive coral Goniastrea aspera in Okinawa, subtropical Japan

The effect of intraspecific contact (Contact) on egg production was examined in the massive coral Goniastrea aspera in Okinawa, subtropical Japan. The contact was non-aggressive without damaging soft tissues each other. Within Contact colonies, polyp volume, polyp fertility (%polyps with gonad), and NE/PV (number of eggs per polyp volume) were significantly smaller in marginal (Mg) polyps without direct intraspecific contact than other polyps, but no difference was found between non-marginal and Mg-Contact (marginal with direct intraspecific contact) polyps. Comparisons of non-marginal polyps (non-marginal and Mg-Contact polyps were combined in Contact colonies) between Non-Contact and Contact colonies showed that fertility and NE/PV were significantly larger in Contact colonies than in Non-Contact colonies, but polyp volume were not different significantly. Further analyses dividing colonies at Non-Contact maturation colony size (60 polyps) revealed that fertility and NE/PV were significantly larger in Contact colonies than in Non-Contact colonies only in the small colonies (<60 polyps), indicating that the intraspecific contact promoted sexual maturation at smaller colony size; one polyped Contact coral was also reproductive. The lack of correlation between polyp volume and NE/PV in the small Contact colonies, and the similarity of NE/PV in non-marginal and Mg-Contact polyps within a colony, suggest that the maturation at smaller size in Contact colonies is realized by reproductive integration of polyps at the colony level. The present results show that size-structured populations such as colonial corals may show phenotypic diversity in key demographic parameters, such as reproductive output, dependent on ecological conditions.     

Mesenterial filaments of the black coral <Emphasis Type="Italic">Cirrhipathes</Emphasis> cfr. <Emphasis Type="Italic">anguina</Emphasis> provide a home to developing nauplii

Inspection of two female colonies of the monopodial black coral Cirrhipathes cfr. anguina from the coral reef of the Marine Park of Bunaken (Indonesia) revealed the occurrence of crustacean developing eggs within the mesenterial filaments of the polyps. Egg diameter, which in the smallest gametes was about 50–60 μm, increased in tandem with embryo development, reaching the value of 170 μm, at the nauplius stage. The attribution to the crustacean taxon was derived from morphological investigations carried out in light and electron microscopy (TEM, SEM) on the eggs and on the embryos removed from them. The final stage of nauplius was characterised by three pairs of appendages: uniramouse antennulae, biramouse antennae and manidibulae. In addition, naupliar eye and caudal setae were also evident. These nauplii were ascribed to the larval stage of an unidentified species. Coral/copepod association could represent a reproductive strategy, put into action by some marine copepods. Incubation within an appropriate host prevents predation by planktotrophic organisms, thus reducing population depletion.     

Strategies for Gardening Denuded Coral Reef Areas: The Applicability of Using Different Types of Coral Material for Reef Restoration

Recreational and other human activities degrade coral reefs worldwide to a point where efficient restoration techniques are needed. Here we tested several strategies for gardening denuded reefs. The gardening concept consists of in situ or ex situ mariculture of coral recruits, followed by their transplantation into degraded reef sites. In situ nurseries were established in Eilat's (Northern Red Sea) shallow waters, sheltering three types of coral materials taken from the branching species Stylophora pistillata (small colonies, branch fragments, and spat) that were monitored for up to two years. Pruning more than 10% of donor colonies' branches increased mortality, and surviving colonies displayed reduced reproductive activity. Maricultured isolated branches, however, exceeded donor colony life span and reproductive activity and added 0.5–45% skeletal mass per year. Forty‐four percent of the small colonies survived after 1.5‐year mariculture, revealing average yearly growth of 75 ± 32%. Three months ex situ maintenance of coral spat (sexual recruits) prior to the in situ nursery phase increased survivorship. Within the next 1.5 years, they developed into colonies of 3–4 cm diameter. Nursery periods of 2 years, 4–5 years, and more than> 5 years have been estimated for small colonies, spat, and isolated branches, respectively. These and other results, including the possible use of nubbins (minute fragments the size of a single or few polyps), are discussed, revealing benefits and drawbacks for each material. In situ coral mariculture is an improved practice to the common but potentially harmful protocol of direct coral transplantation. It is suggested that reef gardening may be used as a key management tool in conservation and restoration of denuded reef areas. The gardening concept may be applicable for coral reefs worldwide through site‐specific considerations and the use of different local coral species.     

Social kin associations and genetic structuring of striped dolphin populations (Stenella coeruleoalba) in the Mediterranean Sea

We investigated hierarchical patterns of genetic subdivision, and assessed kinship within and between social groups of striped dolphins (Stenella coeruleoalba) in the Tyrrhenian Sea. A total of 165 samples were analysed at eight microsatellite DNA loci, including outgroup samples from the Adriatic, Scotland and Spain for population-level comparisons. We found population genetic structure within the Mediterranean basin, including small but significant differentiation between the Adriatic and Tyrrhenian Seas (FST=0.0047, P=0.008), and between putative 'inshore' and 'offshore' (FST=0.0217, P=0.005) populations in the Tyrrhenian Sea. Assessment of kinship within and among 12 association groups showed higher average kinship for females within than between groups, and smaller groups showed higher average kinship. Comparisons of relatedness for both sexes showed a significant difference between males and females, with females more likely to associate with adult kin. Together these data emphasize the importance of the social cohesion of kin in small groups to the structuring of striped dolphin populations in this environment.     

Quantifying Spatial Genetic Structuring in Mesophotic Populations of the Precious Coral Corallium rubrum

While shallow water red coral populations have been overharvested in the past, nowadays, commercial harvesting shifted its pressure on mesophotic organisms. An understanding of red coral population structure, particularly larval dispersal patterns and connectivity among harvested populations is paramount to the viability of the species. In order to determine patterns of genetic spatial structuring of deep water Corallium rubrum populations, for the first time, colonies found between 58–118 m depth within the Tyrrhenian Sea were collected and analyzed. Ten microsatellite loci and two regions of mitochondrial DNA (mtMSH and mtC) were used to quantify patterns of genetic diversity within populations and to define population structuring at spatial scales from tens of metres to hundreds of kilometres. Microsatellites showed heterozygote deficiencies in all populations. Significant levels of genetic differentiation were observed at all investigated spatial scales, suggesting that populations are likely to be isolated. This differentiation may by the results of biological interactions, occurring within a small spatial scale and/or abiotic factors acting at a larger scale. Mitochondrial markers revealed significant genetic structuring at spatial scales greater then 100 km showing the occurrence of a barrier to gene flow between northern and southern Tyrrhenian populations. These findings provide support for the establishment of marine protected areas in the deep sea and off-shore reefs, in order to effectively maintain genetic diversity of mesophotic red coral populations.     

The ecology of the Mediterranean stony coral Cladocora caespitosa (Linnaeus, 1767) in the Gulf of Trieste (northern Adriatic Sea): a 30-year long story

Cladocora caespitosa is an endemic coral of the Mediterranean Sea and an important carbonate bioconstructor that adds 3D complexity to the habitat, thus increasing marine biodiversity. Despite its important role in the ecosystem, the real status of the population along most of the Mediterranean coastline is still poorly investigated and very little is known about the resilience of the species. Using non-destructive visual surveys, colonies of C. caespitosa were investigated by SCUBA diving in 2013 and 2015 at seven sites of the northern Adriatic Sea (southern part of the Gulf of Trieste). Data about colony size, index of sphericity and corallite diameter were collected. Almost all biometrical parameters differed significantly among sampling sites, showing low occurrence of the larger size classes compared to the abundance of small-sized colonies. This pattern of distribution is typical of long-lived organisms. The positively skewed colony size distribution could be due to both a high mortality rate of small colonies unable to reach larger size classes, and to a high fragmentation rate of colonies, related to a strong hydrodynamic forces. The northern Adriatic population of C. caespitosa has previously been investigated by Schiller, who reported size and abundance data of colonies from one site, at a depth range of 2–5?m. We compared these data with our findings from the same sampling site, adding new information about the ecology of C. caespitosa. After a 30-year period, the comparison shows a change in the size distribution of colonies, with a decrease of the small class and an increase of the medium class of colonies. In view of these conclusions, further assessments are required in order to evaluate the trend of the northernmost C. caespitosa population in the Mediterranean Sea.     

Sperm dispersal distances estimated by parentage analysis in a brooding scleractinian coral

Within populations of brooding sessile corals, sperm dispersal constitutes the mechanism by which gametes interact and mating occurs, and forms the first link in the network of processes that determine specieswide connectivity patterns. However, almost nothing is known about sperm dispersal for any internally fertilizing coral. In this study, we conducted a parentage analysis on coral larvae collected from an area of mapped colonies, to measure the distance sperm disperses for the first time in a reef‐building coral and estimated the mating system characteristics of a recently identified putative cryptic species within the Seriatopora hystrix complex (ShA; Warner et al. 2015). We defined consensus criteria among several replicated methods (colony 2.0, cervus 3.0, mltr v3.2) to maximize accuracy in paternity assignments. Thirteen progeny arrays indicated that this putative species produces exclusively sexually derived, primarily outcrossed larvae (mean t_m = 0.999) in multiple paternity broods (mean r_p = 0.119). Self‐fertilization was directly detected at low frequency for all broods combined (2.8%), but comprised 23% of matings in one brood. Although over 82% of mating occurred between colonies within 10 m of each other (mean sperm dispersal = 5.5 m ± 4.37 SD), we found no evidence of inbreeding in the established population. Restricted dispersal of sperm compared to slightly greater larval dispersal appears to limit inbreeding among close relatives in this cryptic species. Our findings establish a good basis for further work on sperm dispersal in brooding corals and provide the first information about the mating system of a newly identified and abundant cryptic species.     

Transfer of intracolonial genetic variability through gametes in Acropora hyacinthus corals

In recent years, the new phenomenon of intracolonial genetic variability within a single coral colony has been described. This connotes that coral colonies do not necessarily consist of only a single genotype, but may contain several distinct genotypes. Harboring more than one genotype could improve survival under stressful environmental conditions, e.g., climate change. However, so far it remained unclear whether the intracolonial genetic variability of the adult coral is also present in the gametes. We investigated the occurrence of intracolonial genetic variability in 14 mature colonies of the coral Acropora hyacinthus using eight microsatellite loci. A grid was placed over each colony before spawning, and the emerging egg/sperm bundles were collected separately in each grid. The underlying tissues as well as the egg/sperm bundles were genotyped to determine whether different genotypes were present. Within the 14 mature colonies, we detected 10 colonies with more than one genotype (intracolonial genetic variability). Four out of these 10 mature colonies showed a transfer of different genotypes via the eggs to the next generation. In two out of these four cases, we found additional alleles, and in the two other cases, we found only a subset of alleles in the unfertilized eggs. Our results suggest that during reproduction of A. hyacinthus, more than one genotype per colony is able to reproduce. We discuss the occurrence of different genotypes within a single coral colony and the ability for those to release eggs which are genetically distinct.     

Organic aggregates formed by benthopleustophyte brown alga Acinetospora crinita (Acinetosporaceae,Ectocarpales)

This work presents the elemental, polysaccharide, and fatty acid compositions of benthic aggregates formed by the filamentous brown alga Acinetospora crinita, which are widely spread on the rocky bottoms of the Mediterranean Sea. The aggregates can be characterized as mineralized centers in which regeneration of nutrients and recycling of dissolved organic matter actively occur and favor the development of an abundant phytoplankton community. Analyses of the stable isotopes of C and N display their marine origin and could provide evidence of the processes that occur inside/outside of the aggregates. The monosaccharide compositions of Adriatic and Tyrrhenian mucilages produced by brown alga A. crinita were quite similar. In particular, the Adriatic sample compositions resembled the average composition of the Tyrrhenian high molecular weight exopolymers, and the observed differences could be ascribed to different degradation stages. The fatty acid patterns found for the aggregates were similar to those observed in the isolated A. crinita algae with variable contributions from embedded diatom species. The bacterial contribution to the fatty acid pool was quite low, most likely due to the known poor conditions for their heterotrophic growth.     

Fungi in Porites lutea: association with healthy and diseased corals.

Healthy and diseased scleractinian corals have been reported to harbour fungi. However, the species of fungi occurring in them and their prevalence in terms of biomass have not been determined and their role in coral diseases is not clear. We have found fungi to occur regularly in healthy, partially dead, bleached and pink-line syndrome (PLS)-affected scleractinian coral, Porites lutea, in the reefs of Lakshadweep Islands in the Arabian Sea. Mostly terrestrial species of fungi were isolated in culture from these corals. Hyaline and dark, non-sporulating fungi were the most dominant forms. Fungal hyphae extended up to 3 cm within the corals. Immunofluorescence detection using polyclonal immunological probes for a dark, initially non-sporulating isolate (isolate # 98-N28) and for a hyaline, non-sporulating fungus (isolate # 98-N18) revealed high frequencies of these in PLS-affected, dead and healthy colonies of P. lutea. Total fungal biomass accounted for 0.04 to 0.05% of the weight of corals in bleached corals and was higher than in PLS-affected and healthy colonies. Scanning electron microscopy revealed the presence of fungi within the carbonate skeleton and around polyps. Fungi appear to be a regular component of healthy, partially dead and diseased coral skeleton.     

On the occurrence of Calycophorae (Siphonophora) in the southern Adriatic and Tyrrhenian Sea: a comparison of the annual cycles off Dubrovnik and Naples

The 1965/66 samples taken monthly at three stations in the Bayof Naples and in the south of Dubrovnik were used to comparethe calycophore fauna of the Tyrrhenian Sea and the Adriatic.Nearly identical calycophore fauna were found, except for threerare Atlantic forms registered only by the deep-sea stationin the Tyrrhenian Sea. Of numerical importance were the neriticspecies Muggiaea kochi, Lensia subtilis, Sphaeronectes gracilisand S. irregularis, and the characteristic open-sea speciesEudoxoida spiralis and the deep-sea species Lensia melton. Theannual numerical abundance maxima in both areas were noted inspring. However, in the Tyrrhenian Sea, the values were twotimes higher, which agrees with the productivity patterns. ¹Prof. Dr Tomo Gamulin died in Dubrovnik on June 4, 1991     

Propagation of the threatened staghorn coral Acropora cervicornis: methods to minimize the impacts of fragment collection and maximize production

Coral reef restoration methods such as coral gardening are becoming increasingly considered as viable options to mitigate reef degradation and enhance recovery of depleted coral populations. In this study, we describe several aspects of the coral gardening approach that demonstrate this methodology is an effective way of propagating the threatened Caribbean staghorn coral Acropora cervicornis: (1) the growth of colonies within the nursery exceeded the growth rates of wild staghorn colonies in the same region; (2) the collection of branch tips did not result in any further mortality to the donor colonies beyond the coral removed for transplantation; (3) decreases in linear extension of the donor branches were only temporary and donor branches grew faster than control branches after an initial recovery period of approximately 3–6 weeks; (4) fragmentation did not affect the growth rates of non-donor branches within the same colony; (5) small branch tips experienced initial mortality due to handling and transportation but surviving tips grew well over time; and (6) when the growth of the branch tips is added to the regrowth of the fragmented donor branches, the new coral produced was 1.4–1.8 times more than new growth in undisturbed colonies. Based on these results, the collection of small (2.5–3.5 cm) branch tips was an effective propagation method for this branching coral species resulting in increased biomass accumulation and limited damage to parental stocks.     

Sperm ultrastructure of a member of the black coral family Aphanipathidae: Rhipidipathes reticulata (Anthozoa, Antipatharia)

Fertile male polyps of three colonies of the black coral Rhipidipathes reticulata (Aphanipathidae) from Togian Islands (Indonesia) have been the source of the sperm investigated at ultrastructural level, in order to compare their organization with that of other members belonging to the family Antipathidae and Myriopathidae. The extension of the studies to a representative of the family Aphanipathidae stresses once more the structural similarity of the male gametes in antipatharians. A sketch of the sperm model reports the similarity and differences in the examined taxa. Among the micro-characters, the cup-like body linked to the pericentriolar apparatus, is exclusive of the antipatharians. Other inclusions concern the more common pro-acrosomal vesicles or the acrosomelike structure observed only in Antipathella subpinnata and Myriopathes japonica. Lipid vesicles are occasionally present. A typical inclusion, the electron-dense content of which has a C-shaped configuration, is restricted to Rhipidipathes reticulata and is associated to the cup-like body or to the mitochondrion.     

Genetic and morphological analyses reveal a complex biogeographic pattern in the endemic barbel populations of the southern Italian peninsula

The Italian peninsula is a biodiversity hotspot, with its freshwater fish fauna characterized by high levels of local endemism. Two endemic fluvio‐lacustrine fishes of the genus Barbus (barbel, family Cyprinidae) have allopatric distributions in the Tyrrhenian and Adriatic basins of Italy. Barbus plebejus inhabits the mid‐ to northern Adriatic basins, while B. tyberinus is widespread in all central‐northern basins draining into the Tyrrhenian Sea. For basins in Southern Italy draining into the southern parts of these seas, there remains a knowledge gap on their barbel populations due to no previous genetic and morphological studies, despite their apparent biogeographic isolation. Correspondingly, this study quantified the presence and distribution of barbels in the Adriatic and Tyrrhenian basins of Southern Italy through genetic and morphological analyses of 197 fish sampled across eight populations. Testing of how local isolation has influenced the evolution and persistence of these populations was completed by examining sequence variation at two mitochondrial loci (cytochrome b and D‐loop) and performing geometric morphometric analyses of body shape, plus measuring 11 morphometric and meristic characters. Phylogenetic and morphological analyses revealed the presence of two genetically distinct lineages that differed significantly from adjacent B. tyberinus and B. plebejus populations. These two new taxa, here described as SI1 and SI2 Barbus lineages, are highly structured and reflect a complex mosaic biogeographic pattern that is strongly associated with the underlying hydrographical scenarios of the basins. The geographic isolation of these basins thus has high evolutionary importance that has to be considered for maintaining endemism.     

The use of coral nubbins in coral reef ecotoxicology testing

While there is an urgent demand to establish reliable ecotoxicological assays for reef corals, there has not been yet an available source material that can supply the high number of colony replicates needed for reliable tests. In past experiments, the major obstacle to obtaining as many fragments as possible had been the damage inflicted to donor colonies by pruning. In this paper, we present the application of coral nubbins, a novel source material for coral ecotoxicology assays. Nubbins from the branching Red Sea coral Stylophora pistillata (n>450) were used for evaluating the impacts of water soluble fractions from a crude oil, an oil dispersant and dispersed oil. Coral nubbins (minute coral fragments in the size of one to several polyps) harvested from a single colony are genetically identical to each other, may be obtained in any quantity needed and whenever research activities demand their use. Several dozens of nubbins can be obtained from a single small branch in branching coral species, a procedure that has minimal impact on donor genotypes. Nubbins production is a low cost procedure and requires limited maintenance space. Results of short and long-term acute ecotoxicological tests are revealed and discussed here, indicating the advantageous use of nubbins as ubiquitous coral material for toxicology assays and physiological studies.     

Sediment rejection by reef corals: The roles of behavior and morphology in Montastrea cavernosa (Linnaeus)

The sediment rejection abilities of colonies of Montastrea cavernosa (Linnaeus) were monitored in a series of field and laboratory experiments. Under conditions of natural sediment deposition, colonies were capable of removing virtually all sediment from their surfaces. Equivalent sections of dead coral skeleton accumulated as much as 345 mg sediment · 25 cm^?2 · day^?1. Laboratory experiments show the sediment rejection process to consist of passive and active phases. Passive movement of sediment off colonies occurs at the time of initial sediment impact. Fine grain size, tall polyps, and convex colonies all enhance passive removal. Active removal involves the behavioral responses of the colony and is independent of sediment type or colony morphology. The presence of appropriate morphologies can increase the proportion of sediment passively removed and reduce the amount of work a colony must perform to keep its surface clear of sediment.