Production,maturity, reproductive cycle and fecundity of small-spotted catshark,Scyliorhinus canicula (Chondrichthyes: Scyliorhinidae) from the northern coast of Tunisia (Central Mediterranean)

Small-spotted catshark Scyliorhinus canicula (Linnaeus, 1758) is confronted since 2000 to a drastic decline of captures in Tunisian northern areas where it formerly was quantitatively and qualitatively very abundant. A study conducted in the area showed that males and females sexually matured between 400–478 mm and 364–471 mm total length (TL), respectively. The largest male and female were 521 mm and 531 mm TL, respectively and weighed 449 and 445.5 g, respectively. There was a not significant relationship total mass vs. TL between males and females. Conversely, the relationship of liver mass vs. TL significantly differed between sexes. Both HSI and GSI values did not present significant differences between males and females. There was a not significant relationship total mass vs. TL between males and females. Both male and female HSI reached high values in adults, significantly higher than those of sub-adults. Both male and female GSI values increased with TL of specimens; additionally, they showed significant differences between juveniles and sub-adults and also between sub-adults and adults. The OSI values were significantly different between juvenile and sub-adult females, and also between sub-adult and adult females. Monthly mean values of adult male and female HSI did not significantly vary throughout the year. Significant monthly changes were recorded in mean values of adult male however no significant changes were observed in monthly mean of GSI and OSI for adult females. Vitellogenic activity and production of egg cases permanently occurred throughout the year. The diameter of largest yolky oocytes ranged from 20.1 to 23.0 (mean = 21.43 ± 1.07) and weighed from 0.64 to 0.90 g (mean = 0.78 ± 0.09). Some measurements were taken on egg cases: length with horns ranged 42.5-55.7 mm with mean = 48.9 ± 2.9, width ranged 13.6–19.7 mm with mean = 16.50 ± 0.9, while the total mass of complete egg case ranged between 1.8 and 4.6 g, with mean = 2.4 ± 0.4. An annual fecundity estimation based on production of egg cases and oocytes during one year counted in adult females, enabled us to consider it between 40 and 240.     

Lipid dynamics in the common torpedo, Torpedo torpedo, from the south eastern Mediterranean

Liver stores the major part of lipids in both mature and immature Torpedo torpedo of both sexes. Fluctuations in the lipid contents of liver, muscles and gonads of immature fish followed the relative change in food intake throughout the year. Hepatic and muscular lipids in mature females diminished before ovulation and were intensively consumed in the last stages of gestation (in pregnant females). After parturition, the recovery of the lipid stores was very rapid due to intensive food uptake. Storing of hepatic lipids in mature males started a little before gonadal maturation and reached a maximum in January and February while gonadal lipids decreased in the same period. Gonadal lipid for both sexes fluctuated in parallel to fluctuations in the GSI. Water content showed significant inverse correlation with lipid content in both liver and gonad tissues, but no significant correlation was recorded in the muscle tissues.     

Bacterial Community Structure of Sediments of the Bizerte Lagoon (Tunisia), a Southern Mediterranean Coastal Anthropized Lagoon

In order to estimate how pollution affects the bacterial community structure and composition of sediments, chemical and molecular approaches were combined to investigate eight stations around the Bizerte lagoon. Terminal restriction fragment length polymorphism (T-RFLP) analysis of PCR-amplified 16S rRNA genes revealed that each station was characterized by a specific bacterial community structure. The combination of this data with those of chemical analysis showed a correlation between the bacterial fingerprint and the pollutant content, principally with hydrocarbon pollution. The composition of the bacterial community of two contrasted stations related to the pollution revealed sequences affiliated to α, β, γ, δ, ε subclass of the Proteobacteria, Actinobacteria, and Acidobacteria in both stations although in different extent. Gamma and delta subclass of the Proteobacteria were dominant and represent 70% of clones in the heavy-metal-contaminated station and 47% in the polyaromatic hydrocarbon (PAH)-contaminated. Nevertheless, most of the sequences found were unaffiliated to cultured bacteria. The adaptation of the bacterial community mainly to PAH compounds demonstrated here and the fact that these bacterial communities are mainly unknown suggest that the Bizerte lagoon is an interesting environment to understand the capacity of bacteria to cope with some pollutants.     

Relative growth and reproductive cycle of the date mussel Lithophaga lithophaga (Linnaeus, 1758) sampled from the Bizerte Bay (Northern Tunisia)

Relative growth and the reproductive cycle of the date mussel Lithophaga lithophaga were studied from September 2002 to October 2003 in the Bizerte Bay (Northern Tunisia). Relationships between shell length and shell width, shell height as well as volume-related variables (shell dry weight, soft tissues dry weight and total wet weight) showed negative allometries in both sexes except for shell width in males (isometry) and shell height in both sexes (positive allometry). The sex ratio was unbalanced, within the length range of 6–48 mm, 49.43 % of the animals were males, 24.39 % females and 26.17 % sexually undifferentiated; within the length range of 49–92 mm, the respective values were 41.19, 52.7 and 6.1 %. Histological investigations as well as analyses of the condition index and the gonadosomatic index (CI and GSI) revealed the presence of a single reproductive cycle per year. Spawning occurred at the end of August and early September and was associated with a decrease in seawater temperature and salinity. A resting phase occurred in winter, coinciding with the lowest water temperatures. Histological examinations of the gonads of a total of 130 specimens revealed only two cases of hermaphroditism. The present study constitutes a useful baseline for a sustainable management of local wild stocks of L. lithophaga.     

Molecular characterization and chromosomal localization of spermatogenesis related sequences in Torpedo torpedo (Chondrichthyes, Torpediniformes)

Cytogenetic data in cartilaginous fishes are currently very inconsistent, considering that the karyotype morphology of only about seventy living species is actually known. Only in the last few years different molecular approaches, first of all physical mapping on metaphase chromosomes, have been used to investigate the cytotaxonomic relationship existing inside this interesting group of vertebrates. The aim of the work was to characterize new molecular chromosomal markers both to discriminate the different chromosome pairs and to distinguish the probable sex chromosomes in the species Torpedo torpedo, since its karyotype does not seem to exhibit heterochromosomes. Evolution of the SRY gene has received considerable attention, mainly because it has been shown to be the sex-determining locus in mammals. The gene is located in the Y chromosome where it normally occurs as a single copy. Using primers taken from the conserved SRY sequences, we characterized these regions at the molecular level and localized them on metaphase chromosomes. The PCR products revealed similar patterns in specimens of both sexes of T. torpedo, but only one fragment of the male amplification product showed a high percentage of identity with human spermatogenesis related genes, SPATA 16, SPATA 18 and UTY. Fluorescent in situ hybridization with these sequences showed the presence of spots at the subtelomeric level of two chromosome pairs in the male and of one pair in the female. Finally, these sequences are particularly useful as chromosome markers to differentiate between the male and the female karyotypes in this species.     

Effects of N and P supply on phytoplankton in Bizerte Lagoon (western Mediterranean)

Enriched bottle experiments were conducted in situ during winter (January and February) and summer (July and August) 2001 to examine the effects of nutrient enrichments (+ N, + P and + NP) on phytoplankton in Bizerte Lagoon, Tunisia. Chlorophyll a (Chl a), ranging from 3.05 μg L⁻¹ in winter to 4.52 μg L⁻¹ in summer, was dominated by the small size-faction (<5 μm) during both seasons. However, the contribution of the large size-fraction (5-200 μm) to Chl a increased from winter (26%) to summer (37%). Similarly, the carbon biomass of the 5-200 μm algae increased during the July/August period that was characterised by the high proliferation of several diatom taxa. In winter, N was the limiting element for phytoplankton growth. Its addition alone (+ N) or with P (+ NP) increased both the <5 μm and 5-200 μm Chl a concentrations. There was no change in the phytoplankton size structure, with the small cells dominating the final algal biomass in all treatments after 5 days. In summer, N and P limited the phytoplankton, but small and large algae exhibited diverse responses to different nutrient enrichments: addition of P increased the Chl a only in the 5-200 μm fraction, the + N treatment enhanced both size classes, and the NP fertilisation mostly stimulated the biomass of large cells. Consequently, the N and P addition in summer was followed by a significant change in the phytoplankton size structure, since both size-fractions contributed equally to the final Chl a biomass. Within the 5-200 μm algal community, various taxa had diverse responses to the nutrient supply during both seasons, leading to a change in the final community composition. The autotrophic flagellates appeared to grow well under N-deficient conditions. In contrast, diatom growth and biomass were mostly stimulated by the N enrichment while dinoflagellates exhibited the highest increase in their growth and biomass with P fertilisation. Our results suggest that the increasing anthropogenic supply of nutrients in the lagoon may influence algal dynamics as well as productivity in different ways depending on the nutrient composition.     

Environmental Quality Assessment of Bizerte Lagoon (Tunisia) Using Living Foraminifera Assemblages and a Multiproxy Approach

This study investigated the environmental quality of the Bizerte Lagoon (Tunisia) through an integrated approach that combined environmental, biogeochemical, and living benthic foraminiferal analyses. Specifically, we analyzed the physicochemical parameters of the water and sediment. The textural, mineralogical, and geochemical characteristics of the sediment, including total organic carbon, total nitrogen, simultaneously extracted metals (SEM), acid volatile sulfides (AVS), chlorophyll a, CaCO₃, and changes in bacterial populations and carbon isotopes were measured. The SEM/AVS values indicated the presence of relatively high concentrations of toxic metals in only some areas. Foraminiferal assemblages were dominated by species such as A. parkinsoniana (20–91%), Bolivina striatula (<40%), Hopkinsina atlantica (<17%), and Bolivina ordinaria (<15%) that cannot be considered typical of impacted coastal lagoons both in Mediterranean and northeast Atlantic regions. The results of this work suggest that Bizerte Lagoon is a unique setting. This lagoon is populated by typical marine species that invaded this ecosystem, attracted not only by the prevailing favorable environmental conditions but also by the abundance and quality of food. The results indicate that the metal pollution found in some areas have a negative impact on the assemblages of foraminifera. At present, however, this negative impact is not highly alarming.     

Temporal variability of abundance and reproductive traits of Centropages kroyeri (Calanoida; Copepoda) in Bizerte Channel (SW Mediterranean Sea, Tunisia)

Abundance and reproductive traits (spawning female, egg production, egg hatching success) and naupliar survival of the dominant calanoid copepod Centropages kroyeri were investigated in relation to biotic (chlorophyll a, phytoplankton and microplankton) and abiotic (temperature and salinity) parameters at a monitoring station located in Bizerte Channel, northern Tunisia. The monitoring was carried out between July 2004 and December 2004, according to two temporal sampling strategies. First, during July, the supposed maximum abundance period of C. kroyeri, daily sampling was conducted. Thereafter, sampling was carried out weekly until the end of the sampling period.The temporal variability of the environmental factors in relation to the reproductive traits of C. kroyeri revealed, first, that temperature is important in controlling the reproduction and dynamics of this species, and second, that the quantity and quality of food available in the field also strongly influence its reproduction. On several sampling dates, the output of certain reproductive traits such as hatching success after 24 h was very weak. It is possible that C. kroyeri produced resting eggs during the study period. Nerveless, the analysis of the phytoplankton during these periods showed that low reproductive activity may have been caused by low phytoplankton quantity and/or by the dominant presence of certain diatom species that are supposed to have an inhibitory effect on the reproductive traits of C. kroyeri, especially on egg hatching success.Among the considered external parameters, temperature and food quality and availability seem to be the most important factors for the reproduction and dynamics of C. kroyeri. Under these biotic and abiotic field conditions, this copepod seems able to modulate the abundance of its population while controlling its reproduction.     

Alexandrium pacificum Litaker sp. nov (Group IV): Resting cyst distribution and toxin profile of vegetative cells in Bizerte Lagoon (Tunisia,Southern Mediterranean Sea)

A high spatial resolution sampling of Alexandrium pacificum cysts, along with sediment characteristics (% H₂O, % organic matter (OM), granulometry), vegetative cell abundance and environmental factors were investigated at 123 study stations in Bizerte Lagoon (Tunisia). Morphological examination and ribotyping of cells obtained from a culture called ABZ1 obtained from a cyst isolated in lagoon sediment confirmed that the species was A. pacificum. The toxin profile from the ABZ1 culture harvested during exponential growth phase was simple and composed of the N-sulfocarbamoyl toxins C1 (9.82 pg toxin cell⁻¹), the GTX6 (3.26 pg toxin cell⁻¹) and the carbamoyl toxin Neo-STX (0.38 pg toxin cell⁻¹). The latter represented only 2.8% of the total toxins in this strain.High abundance of A. pacificum cysts correlated with enhanced percentages of water and organic matter in the sediment. In addition, sediment fractions of less than 63 μm were examined as a favorable potential seedbed for initiation of future blooms and outbreaks of A. pacificum in the lagoon. A significant difference in the cyst distribution pattern was recorded among the lagoon's different zones, with the higher cyst abundance occurring in the inner waters. Also, no correlation due to the specific hydrodynamics of the lagoon was observed in the spatial distribution of A. pacificum cysts and vegetative cells.     

Intra‐annual variation of the spatiotemporal distribution and abundance of Talitridae and Oniscidea (Crustacea,Peracarida) at Bizerte Lagoon (northern Tunisia)

During 1 year from spring 2007 to winter 2008, we conducted four seasonal samplings along a transect in the supralittoral zone of Bizerte Lagoon at Menzel Jmil (Tunisia) to study the intra‐annual variation of Peracarida diversity, Talitridae and Oniscidea. For each season, one transect was studied and quadrates were placed successively from the shoreline to the road backing the shore. Talitridae and Oniscidea were the most abundant taxa in our samples reaching highest densities in summer, when fourteen species were found. The minimum and maximum values of species richness of Talitridae were observed in winter (four species) and summer (eight species), respectively, while for Oniscidea, species richness ranged from one species in winter and five species in spring. Both Talitridae and Oniscidea exhibited intra‐annual variation in their spatial distribution; in that, they moved away from the shoreline in winter and were found near the waterline in the other seasons. ANOVA test shows a high significant correlation between both Talitridae with Suaeda maritima and Oniscidea with the plants S. maritima,Salicornia arabica and Obione portulacoides.     

Annual reproductive cycle,spawning periodicity and sexual maturity of blue runner Caranx crysos (Pisces,Carangidae) from the Gulf of Gabes (Tunisia,Eastern Mediterranean)

From June 2004 to May 2006, the reproductive biology of the blue runner Caranx crysos was studied in the Gulf of Gabes (Southern Tunisia). Of 1668 individuals examined, 777 were females (46.6%) and 891 were males (53.4%). The sex ratio significantly deviated in favour of males (♀:♂ = 0.87 : 1). The difference in the numbers of females and males was significant among size‐classes. However, there was no significant difference between sexes over months and seasons. Monthly changes in the Gonado‐Somatic Index (GSI) showed a rapid increase during summer to very high levels in July (3.51% for males and 2.55% for females) and August (3.47% for males and 2.59% for females) before declining sharply in September (0.58% for females and 0.38% for males). The gametogenesis activity began with a pre‐maturation phase, from 20 May to the first 10 days of July, followed by a pre‐spawning phase, from 20 July to 20 August. From the last 10 days of August to 20 September the gonads were in the ripe and spawning stages. From the end of September to early May, gonads were in the post‐spawning and resting stages. The size at which 50% of the population reached sexual maturity was significantly different in males and females: males attained sexual maturity at fork length FL₅₀ = 210.20, whereas females attained maturity at FL₅₀ = 222.3 mm. Age at maturity was 2.4 years for males and 2.8 years for females.     

Examination of the bioindicator potential of Oncholaimus campylocercoides (Oncholaimidae,Nematoda) from Bizerte bay (Tunisia)

We have tested the use of the free-living nematode Oncholaimus campylocercoides as a bioindicator of petroleum pollution. This was analysed using a polluted-reference sampling strategy for one year in Bizerte bay (Tunisia). We noted a decrease in body dimensions together with many biometric anomalies when organisms are exposed to petroleum compounds. The three first moults and the egg-production phase of specimens of O. campylocercoides from the polluted site TRIC (Tunisian Refining Industries Company) took longer. The egg volume was negatively affected by pollution and the development from egg to juveniles J1 in one side and from juveniles J4 to males from TRIC beach on another side seems to be faster.Our results demonstrated that biometry, life cycle and fecundity are useful indices to biomonitor hydrocarbon pollution in marine ecosystems. Juveniles of the second and third stage and gravid females of O. campylocercoides seem to be the most appropriate bioindicators.     

Seasonal phytoplankton responses to N:Si:P enrichment ratio in the Bizerte Lagoon (southwestern Mediterranean)

This study investigated the role of N, P and Si enrichments on phytoplankton in the Bizerte Lagoon (southwestern Mediterranean Sea, Tunisia) during March, June, August, October and December 2004. Polycarbonate bottles were enriched with different nutrients according to four treatments N:Si:P ratios [+NSi/-P (40:40:1), +P/-NSi (20:20:2,5), +NP/-Si (16:0:1) and +Si/-NP (16:32:1)] and incubated in situ during six days. Chl a and carbon biomass of phytoplankton varied significantly during the course of months, with the highest levels recorded in summer (4-4.4 microg Chl a L(-1) or 1126-1721 microg C L(-1)). Dinoflagellates dominated the initial phytoplankton communities, except in August, when diatoms represented a high fraction of microalgae (48%). Enrichment experiments induced significant increases in Chl a and in the final phytoplankton carbon biomasses. In summer (June/August), Si was the main limiting element for phytoplankton. Diatoms strongly responded to +Si/-NP and +NSi/-P enrichments and dominated the final phytoplankton communities (52-61%) in both treatments. Si played the most important role in the growth and development of diatoms. The biomasses and growth rates of dinoflagellates were significantly stimulated by +P/-NSi and +NP/-Si enrichments. After 6 days, dinoflagellates represented more than 70% of the total phytoplankton biomass in samples subjected to these treatments. Moreover, the addition of +P/-NSi increased the biomasses of several dinoflagellates. This suggests that dinoflagellates were mostly controlled by P availability. Unlike diatoms and dinoflagellates, flagellates showed weak responses to nutrient treatments during only some months of the year. The results showed that phytoplankton dynamics in the lagoon were influenced by nutrients in different manners.     

Distribution of hydrocarbon residues in the sediment from Bizerte lagoon (Northern Tunisia)

The study of hydrocarbon residue by infrared spectrophotometry in the deposits of the Bizerta Lagoon allows us to recognize a polluted zone sector of the Menzel Bourguiba Arsenal. The Eastern part of the lagoon, a zone of "oyster parks" is not affected by pollution. Its average contents of hydrocarbon residues are clearly below the maximum content determined by international regulations. The distribution of hydrocarbon residues in the superficial sediments following three well differentiated zones, reflects the weak hydrodynamism in the lagoon. The vertical variations of this pollution agent contents show the preferential accumulation of the terrigenous material, from lake Ichkeul , in the western part and the weak rate of sedimentation in the eastern zone.     

Bacterial pollution of the littoral waters of the north of Tunisia (Tabarka and Bizerte regions) and of Lake Bizerte

The inshore areas of Tabarka and Bizerte are not submitted to the bacterian pollution. The "Old Port" of Bizerte is mainly contaminated all the year round and sometimes polluted the neighbouring urban beach. The lagoon of Bizerte presents any bacterian pollution and could be adapted to the conchyliculture and to the stabulation of Bivalves, with before some important precautions.     

Random Forest model and TRIX used in combination to assess and diagnose the trophic status of Bizerte Lagoon,southern Mediterranean

A combined multimetric trophic index (TRIX) and the Random Forest (RF) model were used to characterize the trophic status of Bizerte Lagoon. The RF model was used to build a predictive model of chlorophyll a using physicochemical variables (nitrite, nitrate, ammonium, phosphate, oxygen, temperature and salinity) as predictors. The approach is based on physicochemical and biological parameters measured in samples collected twice weekly from January to December 2012 at one representative sampling station located at the lagoon center.The observed TRIX values vary from 5.18 to 6.12, reflecting waters ranging from moderate to poor quality with a high trophic level. The results of the RF model show that the most important predictor of chlorophyll a variation appears to be dissolved oxygen, followed by nitrate and temperature, suggesting a reduced model for this variable. The link between chlorophyll a and observed variables was also studied using numerical models, analyzing the linear statistical correlations and a recently proposed non linear model, the Random Forest. Both methods highlight a high correlation between chlorophyll a and dissolved oxygen as opposed to chlorophyll a and nitrate.     

Occurrence of epibenthic dinoflagellates in relation to biotic substrates and to environmental factors in Southern Mediterranean (Bizerte Bay and Lagoon,Tunisia): An emphasis on the harmful Ostreopsis spp., Prorocentrum lima and Coolia monotis

Harmful events associated with epibenthic dinoflagellates, have been reported more frequently over the last decades. Occurrence of potentially toxic benthic dinoflagellates, on the leaves of two magnoliophytes (Cymodocea nodosa and Zostera noltei) and thalli of the macroalgae (Ulva rigida), was monitored over one year (From May 2015 to April 2016) in the Bizerte Bay and Lagoon (North of Tunisia, Southern Mediterranean Sea). The investigated lagoon is known to be highly anthropized. This is the first report on the seasonal distribution of epibenthic dinoflagellates hosted by natural substrates, from two contrasted, adjacent coastal Mediterranean ecosystems. The environmental factors promoting the development of the harmful epibenthic dinoflagellates Ostreopsis spp., Prorocentrum lima and Coolia monotis were investigated. The highest cell densities were reached by Ostreopsis spp. (1.9 × 10³ cells g⁻¹ FW, in October 2015), P. lima (1.6 × 10³ cells g⁻¹ FW, in June 2015) and C. monotis (1.1 × 10³ cells g⁻¹ FW, in May 2015). C. nodosa and Z. noltei were the most favorable host macrophytes for C. monotis (in station L2) and Ostreopsis spp. (in station L3), respectively. Positive correlations were recorded between Ostreopsis spp. and temperature. Densities of the epibenthic dinoflagellates varied according to the collection site, and a great disparity was observed between the Bay and the Lagoon. Maximum concentrations were recorded on C. nodosa leaves from the Bizerte Bay, while low epiphytic cell abundances were associated with macrophytes sampled from the Bizerte Lagoon. The observed differences in dinoflagellate abundances between the two ecosystems (Bay-Lagoon) seemed not related to the nutrients, but rather to the poor environmental conditions in the lagoon.     

Annual reproductive cycle and condition index of Mactra corallina (Mollusca: Bivalvia) from the north coast of Tunisia

This work aims to examine the reproductive cycle of the bivalve Mactra corallina inhabiting the Gulf of Tunis (Tunisia). Histological analysis indicated that the reproductive cycle is composed of six stages and was synchronous between sexes. Gonadal development began in January. From April to July, an increase in temperature (from 16 to 28°C) and a rise in chlorophyll a (from 3.25 to 6.35 mg/L) marked the period of sexual maturity in both sexes, which was also confirmed by an elevation in condition index (CI). Partial spawning started in April for males and in May for females, peaking in late summer indicating a prolonged reproductive period. The reproductive cycle of M. corallina finished with an inactive stage during autumn. From the total number of animals used in this study, we identified 104 females (52%) and 96 males (48%) corresponding to a sex ratio 1:0.92 that did not differ significantly from a 1:1 ratio.