Temporary Freshwater Research Articles

Terrestrial support of wetland food webs via a dissolved inorganic carbon pathway

AbstractStudies of terrestrial support of aquatic food webs have focused primarily on terrestrial organic matter (t‐OM) directly used by animal and microbial consumers. However, dissolved inorganic carbon (DIC) released from t‐OM might also support aquatic primary producers, a key resource for zooplankton and upper trophic levels. Using 2‐yr 13C measurements of algae, zooplankton, terrestrial detritus, sediments, dissolved and particulate organic matter from six seasonal wetlands, we found that in January when algal concentration was low, zooplankton used t‐OM directly or heterotrophic microbes that decompose t‐OM, but in March and May zooplankton was mainly supported by algae as their basal resources, and the algae used DIC derived from the mineralization and methanogenesis of t‐OM, suggesting the DIC pathway of terrestrial support of aquatic food webs. The decomposition of abundant t‐OM from both bed sediments and water column caused high concentrations of DIC. Despite uptake by algae, about half of DIC produced in January and March ended up as emissions to the atmosphere in the form of CO2. This finding revealed the dual roles of t‐OM in maintaining the productivity and stability of aquatic food webs and in contributing to global carbon emissions. This duality poses challenges for simultaneously mitigating carbon emissions and conserving biological communities in seasonal wetlands. Finally, wide seasonal differences in δ13C in DIC (−12.4‰ to 6.7‰) were observed, mainly driven by air–water CO2 exchange and photosynthesis, suggesting that 13C may be a powerful tool to investigate carbon cycling in shallow, temporary freshwater ecosystems that are widespread but understudied.

Love story or fling? The first observation of Vorticella sp. (Ciliophora: Peritrichia) on the freshwater snail Gyraulus rossmaessleri (Auerswald, 1852)

Love story or fling? The first observation of Vorticella sp. (Ciliophora: Peritrichia) on the freshwater snail Gyraulus rossmaessleri (Auerswald, 1852)

The use of 18S rRNA for identification of the first record of Tadpole Shrimp Lepidurus apus (Linnaeus, 1758) from Jordan

Abstract. Abushattal S, Alnasarat H, Alnaimat SM, Eid E. 2024. The use of 18S rRNA for identification of the first record of Tadpole Shrimp Lepidurus apus (Linnaeus, 1758) from Jordan. Biodiversitas 25: 1223-1229. The tadpole shrimps are living fossils that survive in temporary water pools and are considered benthic organisms that swim and feed on tiny organisms. Two genera of tadpole shrimps exist worldwide: Triops and Lepidurus, which share similar ecological niches and habitats but never coincide. The occurrence of the tadpole shrimp Lepidurus apus in Jordan was confirmed for the first time using the 18S rRNA Gene-Based PCR Assay. The experimental approach encompassed the amplification of biological specimens utilizing a universal amplicon measuring approximately 1,500 base pairs (bp), specifically targeting the 18S ribosomal RNA (rRNA) gene. Subsequently, the acquired sequences underwent scrutiny through BlastN search against the GenBank database for validation and comparative analysis. Genetic analysis was needed to define the species accurately, especially with tadpole shrimp's generally high morphological variability. This species was recorded in a temporary freshwater pool in the desert of Al-Jafr in Ma’an governorate in southern Jordan. The DNA analysis showed a 100% identical sequence of the 18S rRNA gene with Lepidurus apus sequences. This record is important due to the ecological role of the tadpole shrimps as filter feeders, consuming algae, bacteria, and other small particles of organic matter, and as a food source for various fish, birds, and other aquatic animals. In addition, it will pave the way for future research to identify its economic importance, such as its role in mosquito control and its potential role in aquaculture. Finally, tadpole shrimp are sensitive to environmental changes and could be used to indicate climatic changes and habitat degradation. This is mainly due to their sensitivity to changes in water quality, including temperature, pH, and chemical composition, and their niche requirements for survival and reproduction. In addition, monitoring tadpole shrimp distribution and comparing it with historical records can reveal patterns of climate-induced range shifts.

Lovers in strange places: amphibian calling and amplexus detected in tidal mangrove creeks after rainfall

Temporary freshwater lenses can form in saline environments after rainfall, providing essential resources for species including drinking water and dispersal routes. However, there is limited indication that these lenses can also be used for breeding. Herein, we provide evidence of the green and golden bell frog, Litoria aurea, performing breeding activities, including calling and amplexus, in tidal mangrove creeks on Kooragang Island, NSW, Australia. Our findings suggest that these creeks intermittently phase into a low salinity state after the influx of fresh water from rainfall, forming temporary freshwater lenses that can be exploited before the creeks revert to a saline state. These lenses had salinities (1.4 ppt and 4.5 ppt) within the tolerance limit of L. aurea tadpoles, although we are unsure whether oviposition took place and offspring survival to metamorphosis was achieved. It is possible that anthropogenic disturbances to hydrology on Kooragang Island have benefited L. aurea by restricting tidal influences in mangrove creeks, prolonging the duration of freshwater lenses.

Temporal variation and spatial factors affecting zooplankton communities in freshwater rock pools

Most ecological studies have focused on one snapshot in a single location and the same taxonomic group. However, relatively few studies have explored different taxonomic groups over multiple periods, particularly in ephemeral Neotropical ecosystems. This study investigates the seasonal dynamics of zooplankton communities and their responses to environmental variables in temporary pools in the Seasonally Dry Forest Ecosystem, Pernambuco, Brazil. We examined the seasonal fluctuations in species abundance, diversity, and evenness of Cladocera, Copepoda, and Rotifera in five temporary rock pools. Although these pools are meters apart, they vary considerably in size and drought sensitivity, leading to distinct seasonal dynamics mediated by water availability. We asked (i) how do species abundance, diversity, and evenness vary seasonally? (ii) Do these seasonal shifts differ between Cladocera, Copepoda, and Rotifera? And (iii) How do water environmental variables and spatial distance of pools influence zooplankton species composition? We hypothesized that Cladocera and Copepoda are positively affected by the rainy season, but Rotifera is mostly common in the dry season. Our findings underscore seasonality as the primary driver influencing the abundance and composition of zooplankton communities. We observed higher abundance of Cladocera and Rotifera during the post-rainy season, but Copepoda was dominant in the rainy season, leading to a strong seasonal variation in species evenness. These differences among groups highlight the importance of considering biological differences when understanding the ecological drivers of temporary freshwater ecosystems.

Experimental warming and drying act independently on developmental responses for two amphibian species

AbstractClimate change is affecting freshwater habitats worldwide through multiple, interacting pressures that influence the survival, distribution, and persistence of freshwater organisms. Temporary freshwater habitats, or those that dry seasonally, can exhibit wide‐ranging variability in temperature and drying regimes. Aquatic organisms with complex life cycles (e.g., aquatic and terrestrial life stages) often rely on temporary freshwaters to complete their life cycles and have specific adaptations to cope with these dynamic environments. However, climate change is altering the temperature and drying regime of many temporary freshwater habitats worldwide, often resulting in warmer habitats available for shorter durations of time. The ways in which climate‐mediated changes to temporary freshwaters affect development and life history of aquatic organisms, particularly those with complex life cycles, remains poorly understood.We investigated larval development, growth, survival, and relationships among them for two amphibian species, wood frogs (Lithobates sylvaticus) and spring peepers (Pseudacris crucifer), in response to warming and drying in a mesocosm experiment. Over 13 weeks in the spring and summer of 2019, we manipulated temperature and water levels in 48 outdoor pond mesocosms to produce four treatments: control (ambient conditions), drying alone, warming alone, and drying + warming. We predicted reduced survival, body size, and time to metamorphosis for both species in response to warming and drying, with the effect being stronger on wood frogs due to their more specialised habitat use and shorter breeding period.Warming treatments were on average 2°C higher than controls, and drying treatments decreased water depth by 2.5 cm each week. We found that warming temperature shortened larval time to metamorphosis, and drying decreased body size at metamorphosis. Surprisingly, we also found that, across all treatment groups, individuals with early metamorphosis tended to be larger in size and from mesocosms with generally higher survival.Our study indicates that warming and drying may act independently on different developmental responses for amphibians. Our results echo recent calls for a better understanding of the nuanced, often unpredictable, relationships among climate‐mediated environmental pressures and the developmental traits they influence, particularly for organisms with complex life cycles that depend upon dynamic habitat for growth and survival. Understanding these relationships and the mechanisms that drive them will be key to predicting the response of freshwater organisms to climate change.

The conservation value of freshwater habitats for frog communities of lowland fynbos.

Amphibians are more threatened than any other vertebrate class, yet evidence for many threats is missing. The Cape lowland fynbos (endemic scrub biome) is threatened by habitat loss, and natural temporary freshwater habitats are removed in favour of permanent impoundments. In this study, we determine amphibian assemblages across different freshwater habitat types with special attention to the presence of invasive fish. We find that anuran communities differ primarily by habitat type, with permanent water habitats having more widespread taxa, while temporary water bodies have more range restricted taxa. Invasive fish are found to have a significant impact on frogs with toads most tolerant of their presence. Temporary freshwater habitats are a conservation priority in the area, and their amphibian assemblages represent endemic taxa that are intolerant of invasive fish. Conservation of a biodiverse amphibian assemblage in lowland fynbos areas will rely on the creation of temporary freshwater habitats, rather than a northern hemisphere pond based solution.

New leptestherid clam shrimps (Pancrustacea: Branchiopoda: Spinicaudata: Leptestheriidae) from peninsular India.

Spinicaudatan clam shrimps are an evolutionarily ancient lineage restricted to temporary freshwater pools. Use of classical morpho-taxonomic approaches alone have led to some issues in the taxonomy of this group, which are now being resolved through integrative taxonomy. Here, we describe two new leptestherid spiny clam shrimps Leptestheria chalukyae sp. nov. and Leptestheria gomantaki sp. nov. from peninsular India based on their unique morphological characters and distinct phylogenetic position. We also re-describe Leptestheria nobilis and present an overview of the morphological characters of all the Indian leptestherids. Most of the conventional taxonomic characters appear to overlap among all the Indian species, although the combination of occipital condyle shape and the cercopod marginal spines arrangement in combination, appear to be useful in separating leptestherid species.

Influence of rainfall events on zooplankton community characteristics and feeding habits in estuarine–coastal environments

In temperate estuaries, rainfall causes environmental fluctuations, such as salinity and suspended particulate matter (SPM), and can affect zooplankton distribution patterns. This study focused on the effect of temporary freshwater inflow on the composition of major zooplankton species and changes in their ecological status in the Seomjin River estuary before (June) and after (August) rainfall in 2018. Environmental data were collected from 14 and 15 stations before and after rainfall, respectively. All factors except for chlorophyll-a (Chl-a) concentration differed significantly before and after rainfall (p<0.05), and a salinity gradient extended to Yeosu Bay from Gwangyang Bay. Zooplankton abundance decreased significantly after rainfall. There was a high correlation between indicator species abundance and environmental factors after rainfall (correlation coefficient: 0.7521); however, the indicator species and environmental factors did not exhibit a significant correlation with salinity before rainfall. In terms of feeding habit composition, the carnivore proportion showed a significant decrease after rainfall compared to before rainfall (p<0.001), while the particle feeder proportion showed a significant increase after rainfall compared to before (p<0.001). In particular, Corycaeus spp. contributed significantly to the decrease in carnivore abundance after rainfall. Among the particle feeders, Copepodites significantly increased in abundance after rainfall. Carnivore abundance was negatively correlated with salinity, and particle feeder abundance was positively correlated with potential prey sources (SPM and Chl-a concentration), suggesting that particle feeders respond to the food-rich environment after rainfall.

Hidden diversity of Chlorococcum (Chlorophyta) in a shallow temporary freshwater lake: description of Chlorococcum szentendrense sp. nov

ABSTRACT Chlorococcum is a coccoid green algal genus, which contains almost 50 species. The genus is polyphyletic, but in a traditional sense, the cells are coccoid and non-motile with a cup-shape chloroplast in the vegetative stage, while the spores have chlamydomonad appearance. They are distributed worldwide mainly in terrestrial habitats, and the biotechnological potential of several strains has been reported. In this study, three new green algal strains from a shallow, temporary freshwater lake in Hungary are characterized using microscopic (light and transmission electron microscopy) and DNA-based methods (phylogenetic analysis of the ribosomal ITS region, the 18S ribosomal RNA and rbcL genes, and ITS secondary structure comparison). Based on the obtained results, one of the three new isolates is considered to represent a new species, which is described here as Chlorococcum szentendrense sp. nov.

Contrasting biodiversity and food web structure of three temporary freshwater habitats in a tropical biodiversity hotspot

Abstract In many biomes, a variety of different small freshwater habitats, such as pools and phytotelmata can occur together in the same habitat matrix. However, both the biodiversity and the functioning of these ecosystems remain poorly known. Three freshwater habitat types in a tropical West African biodiversity hotspot were studied. The study demonstrated that animal communities in water‐filled tree holes, temporary ponds and granite rock pools were strongly differentiated with exclusive faunas despite their spatial proximity and similar disturbance in the form of seasonal drought. In particular, granite rock pools stood out with a high gamma diversity. Rock pools were also functionally different from the other two habitat types. The three habitats had contrasting predator assemblages, a differential reliance on primary production and different concentrations of available nutrients. The work illustrates that the biodiversity and functions of small temporary freshwater habitats can be strongly differentiated. This shows the unique and potentially complementary roles that these habitats can fulfil in mediating fluxes of energy and nutrients and preserving aquatic biodiversity in landscapes. Temporary aquatic habitats are typically overlooked in conservation policy and local management plans despite being threatened by habitat transformation. In addition, livestock can threaten savanna ponds, wood exploitation can threaten tree holes, and mining and sediment disruption can threaten rock pools. Hence, better knowledge about the ecological functioning of these ecosystems is vital to the implementation of effective conservation strategies.

The impact of salinity on a saline water insect: Contrasting survival and energy budget

Water salinity is a major driver of aquatic insects’ distribution. Saline species are usually generalists with high survival and performance at both low and high salinity levels. Yet, costs of high salinity may be underestimated as these are most often measured in terms of larval life history traits, while effects of larval stressors may only be detectable when looking at physiological traits and traits in the adult stage. Here, we assessed the lethal and sublethal physiological effects of embryonic and larval exposure to a range of salinity levels in the damselfly Lestes macrostigma, both during and after metamorphosis. This species inhabits temporary freshwaters where salinity increases during the drying phase. Salinity had no effect on egg hatching success within the range 2–9.5 g/L sea salt (conductivity range 3.45–14.52 mS/cm). With increasing salinity (up to 16 g/L, 23.35 mS/cm), growth rate decreased and larvae took longer to emerge and did so at a smaller size. Larval survival to metamorphosis increased with salinity up to 8 g/L (12.45 mS/cm) and then declined at 16 g/L. Exposure to salinity in the larval stage had no effect across metamorphosis on both the adult thorax muscle mass and flight performance, and the investment in immune function. Increasing salinity in the larval stage also had no effect on the energy available but increased the energy consumption in the adult stage, resulting in a lower net energy budget. These negative sublethal effects of increasing salinity hence bridged metamorphosis and contrasted with the mortality data, suggesting that the higher mortality at the low salinity levels selected for larvae with the best body condition. Our results highlight the importance of taking into account other life-history and physiological traits, besides mortality, ideally across different life stages, to better understand and predict consequences of increasing salinization on freshwater insects.

The Suspected Contradictory Role of Parental Care in the Adaption of Planktonic Calanoida to Temporary Freshwater

Calanoida have the highest number of species among Copepoda in marine plankton, but not in fresh water, where the greatest number are Cyclopoida. Freshwater Cyclopoida also live in more freshwater sites than Calanoida. This could be a consequence of an invasion of freshwater by marine Cyclopoida before Calanoida. Similar to Cyclopoida, but different from marine Calanoida, freshwater Calanoida females produce egg sacs and care for eggs. This strategy is common among all freshwater plankton, suggesting that the evolution of parental care is an obliged adaption to conquer fresh water. Calanoida, different from Cyclopoida, survive adverse conditions as resting eggs. This life-cycle constraint obliges eggs to survive their mother’s death and wait in the benthos for a certain period. The necessity of completing embryonic development and the hatching of eggs far from the mother’s protection may be responsible for the relatively lower evolutionary success of Calanoida in fresh water compared to Cyclopoida (which rest as juveniles, thus protecting eggs in any moment of their development). Therefore, the brooding of eggs appears to be the obliged solution for Calanoida’s final establishment in fresh water, but the dispersion of eggs on the bottom after the mother’s death and during the rest period is probably the weak point in Calanoida’s competition with Cyclopoida.

Aquatic beetles influence colonization of disparate taxa in small lentic systems.

Structure of natural communities is shaped by both abiotic characteristics and the ongoing processes of community assembly. Important to this process are the habitat selection behaviors and subsequent survival of colonists, both in the context of temporal changes in the abiotic characteristics and priority effects driven by earlier colonists. Aquatic beetles are prevalent in temporary freshwater systems, form speciose assemblages, and are often early colonists of temporary ponds. While beetles have the potential to influence community structure through post‐colonization interactions (predation and competition), our goal was to determine whether the presence of beetle assemblages (versus patches without beetles) influences the colonization and oviposition of a diverse group of animals in a naturally colonized experimental landscape. We established mesocosms that either contained existing beetle assemblages or contained no beetles and assessed abundances of subsequent colonists. Treefrogs, Hyla chrysoscelis, and mosquitoes, Culex restuans, both deposited fewer eggs in patches containing beetle assemblages, while two beetles, Copelatus glyphicus and Paracymus, colonized those patches at lower rates. One beetle, Helophorus linearis, colonized patches containing beetle assemblages at higher rates, while two beetles, Berosus infuscatus and Tropisternus lateralis, exhibited no colonization differences between treatments. Overall, there were no differences in the assemblage structure or richness of beetles that colonized patches. Our results illustrate the importance of species‐specific habitat selection behavior in determining the species composition of habitat patches, while emphasizing the role of priority effects in influencing patterns of community assembly. Habitat selection in response to abiotic and biotic characteristics of habitat patches can potentially create greater spatiotemporal niche separation among the numerous, often closely related species (phylogenetically and trophically), that can be simultaneously found in similar patches across landscapes.

Agricultural and urban delivered nitrate pollution input to Mediterranean temporary freshwaters

Nitrate dual stable isotopes (δ15NNO3 and δ18ONO3) have proven to be a powerful technique to trace nitrate sources and transformations in freshwater systems worldwide. However, most studies have focused on perennial systems, and less is known about intermittent ones. The impacts of intensive agricultural practices and wastewaters in Doñana (SW Spain), an iconic Mediterranean temporary wetland protected as a UNESCO World Heritage Site, were quantified using stable isotope mixing models in a Bayesian framework under different denitrification scenarios. We aimed to identify the main nitrate sources and transformation processes in surface waters of interconnected temporary streams, ponds and marshes, and link them with the main human pressures in the watershed (e.g. intensive fruticulture, urban wastewaters). We measured nitrate (NO3−) concentrations and stable isotopes (δ15NNO3 and δ18ONO3) in water samples collected during different periods over two years (2015–2016). Most sites showed coupled increases of nitrate isotopic values (δ15NNO3 and δ18ONO3), which were higher than reference values of any possible sources (e.g. synthetic/organic fertilizers and wastewaters), indicating fractionations typical of denitrification processes. The main nitrate sources to the watershed were linked to agricultural practices and the use of synthetic fertilizers, but further investigations in other transformation processes that occur simultaneously should be evaluated. These results highlight an important nitrate removal capacity (i.e. denitrification) of the system, which may positively contribute to natural resilience against eutrophication. However, given the high intra and interannual hydrological fluctuations of Mediterranean aquatic systems, future studies on the relative contribution of nitrate sources and processes should increase spatio-temporal resolution of water sampling, and include measurements of groundwater and interstitial water as well as surface water.

Influence of morphometric and geographic factors on plant community composition of temporary freshwater rockpools in the Maltese Islands

ABSTRACT We investigated the influence of 6 morphometric and geographic factors on the structure and composition of plant communities in 56 temporary freshwater rockpools from 7 pool landscapes in the Maltese Islands. The factors considered were water depth, sediment depth, surface area, shading, distance from the coast, and elevation above sea level. Coverage data for 30 plant taxa in 3 functional forms (aquatic, amphibious, and terrestrial) were collected during one hydrological cycle. The relationships between pool morphometry, pool location, and community composition were tested using multivariate analyses. The contribution of spatial variables to variation in community composition across pools was examined using principal coordinate analysis of neighbour matrices (PCNM). The results suggested that pool morphometry varied both within and across landscapes, reflected in similar differences in community composition. Moreover, 47.6% of the explained community variation in the whole dataset was contributed exclusively by space predictors, suggesting that pool “location” is an important determinant of community composition. The morphometric property that exerted the highest influence on the dominance of a given plant functional form was the pool surface area, with greater surface areas promoting the presence of amphibious species relative to obligate hydrophytes. Conversely, the presence of specific species was mainly controlled by water depth and sediment depth. The most influential predictors of community composition varied across landscape, suggesting that pool location can be an important determinant of community structure and composition, and pool morphometry would favour certain functional forms and species over others.

Warming interacts with inundation timing to influence the species composition of California vernal pool communities

Climate change has been associated with alterations in temperature and precipitation regimes. Increased temperatures can change the structure and function of aquatic communities through both biotic and abiotic interactions. Many warming experiments have been conducted in permanent freshwater systems, but few in temporary freshwater systems that are dependent on precipitation patterns. Temporary aquatic communities are influenced by hydro-regime characteristics such as the hydroperiod length, magnitude, frequency, and timing of inundation, which may interact with temperature changes. Here, we used mesocosms to test for the effects of warming and inundation timing on the community composition of California vernal pools. California vernal pools are temporary wetlands, which provide habitat for many endemic and endangered species during the aquatic phase. Total species abundance was higher in both warm and late inundation treatments, while higher species richness levels in late treatments were due to the rapid emergence and colonization of species compared to early treatments. Heating increased the abundance of ubiquitous passive and active dispersers, including mosquitos, but decreased the abundance of vernal pool endemic passive dispersers. We also found that significant interactive effects between warming and inundation timing led to differences in community composition. These findings suggest that vernal pool endemic passive disperser species may decline due to increased temperatures and later inundation; however, the extent of such risk will likely be influenced by species-specific life-history traits such as bet-hedging strategies. Conservation efforts should work to identify the species most at risk to a changing climate.

Curculigo sabui sp. nov. (Hypoxidaceae), a new species from Balaghat Ranges of Maharashtra, India

A new species, Curculigo sabui S.P.Gaikwad & Gore is described from margins of temporary fresh water streams in Yedshi‐Ramling Wild Life Sanctuary of Balaghat Ranges of Maharashtra, India. Morphologically it resembles Curculigo janarthanamii Gore & S.P.Gaikwad and C. multiflora Zimudzi but differs in having elliptic‐lanceolate to obovate (20–52 × 3.0–4.5 cm) leaf lamina, flowers three to five in umbel‐like racemes or sometimes solitary, seed coat with irregular striations and somatic chromosome number 2n = 36. A detailed morphological description with line drawings, photographs and comparative account of the species is provided here.

Phytoplankton assemblages and community structure in tropical temporary freshwater ponds in sub-Saharan Africa1.

Anthropogenic and climatic factors are important determinants of algal distribution in aquatic systems. This study aimed to identify temporary pond-groups of phytoplankton assemblages and to establish the interactions between environmental variables and phytoplankton structure. Phytoplankton samples were collected and preserved with 5% formalin; water samples for chemical analysis were taken in August and September 2007, 2008, and 2009 in 86 ponds randomly selected in the Sahelian sector and the Sudanian sector (protected and unprotected areas) of Burkina Faso. Plankton species were examined using light microscopy and identified using standard methods. A Fuchs-Rosenthal chamber was used for algal cell counting. NMDS ordination, indicator species analysis and canonical analysis were performed using Pc-Ord and CANOCO. Kruskal-Wallis and Mann-Whitney U-tests using the software Statistica were performed to compare parameters between pond-groups. Three pond-groups were identified: the Sahelian pond-group, dominated by Lyngbya martensiana and Pseudanabaena constricta; the protected area pond-group, dominated by Euglena caudata and Trachelomonas raciborskii; and the unprotected area pond-group, dominated by Closterium venus and Euglena proxima. At P<0.05, species composition and abundance in algal groups were mostly associated with nitrate, pH, dissolved oxygen, and conductivity. The Sahelian and the Sudanian sectors were different in pH, water transparency, and species richness. Univariate and multivariate analyses illustrate that variations in the physicochemical properties of water and algal structure followed climatic and anthropogenic gradients. The classification of ponds into algal associations demonstrated that algal assemblages may be used as reliable indicators of habitat conditions.

Middle Jurassic charophytes from southern Tunisia: Implications on evolution and paleobiogeography

The Middle Jurassic section from the Kef El Anéba in southern Tunisia, near the Beni Kheddache area, provided exceptionally species-rich charophyte assemblages that are studied here for the first time. The Krachoua Formation yielded a Bajocian assemblage composed of Porochara fusca, P. schudackii nov sp., P. kimmeridgensis, P. westerbeckensis, Auerbachichara saidakovskyi, Stenochara sp., Stellatochara aff. subsphaerica, Aclistochara longiformis and Aclistochara africana. A second assemblage from the Krechem El Miit Member (Foum Tataouine Formation), of Late Callovian age, includes Auerbachichara tataouinensis nov. sp., Porochara kimmeridgensis, P. westerbeckensis, P. douzensis, P. obovata var. gondwanensis nov. var. and Mesochara voluta. Taphonomic and paleoenvironmental data suggest autochthony of these two assemblages in a brackish lagoon and a temporary freshwater pond from a littoral environment, respectively. In addition, these floras suggest that Middle Jurassic charophyte biodiversity formed an evolutionary bridge between Upper Triassic charophyte floras dominated by genera Stellatochara, Stenochara and Auerbachichara and Upper Jurassic floras dominated by Porochara, Aclistochara and Mesochara. Furthermore, the biogeography of Middle Jurassic charophytes from southern Tunisia provides further arguments to support the hypothesis of significant biological exchanges between Laurasian and Gondwanan islands, indicating that Peri-tethyan biogeography remained relatively uniform during the Middle Jurassic.