Vitelline coat of Unio elongatulus: III. Glycan chain analysis of the 220- and 180-kD components by means of lectins

Lectins of different binding specificity were used to analyze the oligosaccharide chains of the 220- and 180-kD proteins of the Unio elongatulus egg vitelline coat (vc). The lectins ConA and RCA₁ reacted with both glycoproteins, and four other lectins reacted with one or other vc components. The lectin from Galanthus nivalis, which recognizes terminal mannose residues of N-linked high mannose type oligosaccharide chains, bound specifically to the 180-kD protein. Binding sites for this lectin were found throughout the vc of the differentiating oocyte and the mature egg. Lectins specific for the O-linked oligosaccharide chains, such as AIA and PNA, reacted only with the 220-kD protein species. Binding sites for these lectins were found only in the crater region. The presence of fucosyl residues on the glycan chains was investigated with lectins from Lotus tetragonolobus and Aleuria aurantia. The latter was positive on both glycoproteins, whereas LTA was only positive to the 220-kD species. The binding sites of both these lectins were in the same areas as those of PNA and AIA. These results suggest that while the 180-kD protein is part of the entire vc structure, the 220-kD protein is prevalently accumulated in the crater region. Since this is where sperm recognition and interaction take place, it has been suggested the 220-kD protein acts as a ligand molecule in the sperm-egg interaction. © 1995 Wiley-Liss, Inc.     

Some common properties of lectins from marine algae

Twelve kinds of lectins isolated from four species of marine algae, Boodlea coacta (Chlorophyta) and Hypnea japonica, Carpopeltis flabellata and Solieria robusta (Rhodophyta), were compared for their chemical and biological properties. These lectins were proteins or glycoproteins, similar to terrestrial plant lectins. However, unlike most terrestrial plant lectins, they had a small molecular size (4,200 to 25,000 daltons), were mostly monomeric, and had no affinity for monosaccharides. They strongly agglutinated trypsin-treated rabbit erythrocytes, and their activities commonly were inhibited by glycoproteins bearing N-glycans. From hemagglutination-inhibition tests with various glycoproteins and related compounds, it was found that B. coacta lectins recognize high-mannose N-glycans; H. japonica lectins complex N-glycans, and C. flabellata and S. robusta lectins recognize both types of N-glycans.     

Copepod post-embryonic durations: pattern,conformity, and predictability. The realities of isochronal and equiproportional development,and trends in the opepodid-naupliar duration ratio

A collation of post-embryonic durations for freshwater and marine calanoid, cyclopoid and harpacticoid copepods is provided, and examined for patterns and conformities which may be of predictive value. Most of the analysis concerns calanoids. Only the genus Acartia exhibits evidence of equal stage duration (isochronality). Accordingly, isochronal development must be rejected as a general pattern in copepods — with various implications to the study of their production. Conversely, relative stage durations are surprisingly comparable in a wide range of copepods across a broad range of temperatures and food levels. A loose consistency is evident among copepod species generally, (interspecific equiproportionality), but within given genera, a striking regularity, here termed intergeneric equiproportionality (IGE) is evident. Patterns of IGE are consistent with selectively adaptive life-history traits, and IGE thus offers predictive prospects which are of both quantitative and qualitative (heuristic) value. Empirical support which exists for IGE among marine calanoids suggests that in contrast to isochronality and interspecific equiproportionality, IGE is indeed a real, and ultimately quantifiable feature of copepod development. Within calanoids, the ratio of total copepodid to total naupliar duration (Dc/Dn) appears independent of temperature, shows little relationship to adult body mass, but is inversely related to food supply, markedly so in freshwater forms. In this context, changes in the ratio are attributable largely to influences of food supply upon copepodid development times: naupliar durations appear relatively independent of food concentrations during development. Considerably higher Dc/Dn ratios in freshwater than in marine calanoids conceivably arise from selective influences of different nutritional conditions and size-selective predation pressures in freshwater and marine environments. In the generally high predation environment of freshwaters, acceleration of naupliar development potentially reduces the vulnerability of these smaller stages to size-selective tactile predation, while larger copepodid instars are able to reduce the opposing size-selective influences of visual planktivores by virtue of their escape responses to suction attack. Within the spectrum of copepod life histories and development schedules, ecologically consistent trends of this nature await formal recognition. The present recognition of IGE is a modest initiative in this quest. Both naupliar and copepodid durations are inverse monotonic functions of temperature. Several mathematical expressions which account for this temperature-duration response are provided. The response envelope is much tighter for naupliar than copepodid durations. Predictability of temperature-duration responses accordingly declines ontogenetically from egg to naupliar to copepodid stages.     

Screening and preliminary characterization of hemagglutinins in Vietnamese marine algae

Extracts from 44 species of Vietnamese marine algae, including 15 Chlorophyta, 18 Rhodophyta and 11 Phaeophyta species, were examined for hemagglutination activity with a variety of different animal and human erythrocytes that were untreated or treated with enzymes. Almost all extracts showed activity toward at least one type of erythrocytes, although those from three Chlorophyta and two Rhodophyta species showed no hemagglutination with any type of erythrocytes examined. Strong activity was detected in extracts from two Chlorophyta (Anadyomene plicata and Avrainvillea erecta) and four Rhodophyta species (Gracilaria eucheumatoides, Gracilaria salicornia, Kappaphycus alvarezii, and Kappaphycus striatum) with enzyme-treated rabbit and sheep erythrocytes. The hemagglutinins of seven Chlorophyta and eight Rhodophyta species were examined for sugar-binding specificity, pH- and temperature-stability, and divalent cation-independency of hemagglutination using ammonium sulfate-precipitates prepared from their extracts. In a hemagglutination-inhibition test with various monosaccharides and glycoproteins, none of the hemagglutinins had affinity for monosaccharides, except the Codium arabicum and Gracilaria euchematoides hemagglutinins, whose activities were inhibited by both N-acetyl-d-galactosamine and N-acetyl-d-glucosamine. On the other hand, all of the hemagglutinins activities were inhibited by some glycoproteins. The inhibition profiles with glycoproteins were different depending on hemagglutinin species, and suggest the presence of lectins specific for high mannose N-glycans, complex N-glycans, or O-glycans. The activities of these algal hemagglutinins were stable over a wide range of pH and temperature, and independent of the presence of divalent cations. These results indicate that Vietnamese marine algae are a good source of novel and useful lectins.     

A new screening for hemagglutinins from Vietnamese marine macroalgae

Aqueous extracts from 42 species of Vietnamese marine macroalgae, including 17 Chlorophyta, 22 Rhodophyta, and three Phaeophyta species, were examined for hemagglutination activity using native and enzyme-treated different animal and human erythrocytes. All extracts agglutinated at least one type of erythrocytes tested. Strong activity was detected in extracts from four Chlorophyta (Caulerpa serulata var. boryana, Caulerpa sertularioides f. longipes, Halimeda velasquezii, and Halimeda discoidea) and two Rhodophyta species (Gelidiella acerosa and Titanophora pulchra) with enzyme-treated rabbit and horse erythrocytes. The hemagglutinins of some active species were examined for sugar-binding specificity, pH, temperature stability, and divalent cation independency using ammonium sulfate precipitates prepared from their extracts. In a hemagglutination–inhibition test with various monosaccharides and glycoproteins, none of the hemagglutinins had affinity for monosaccharides. The activity of the hemagglutinins was inhibited by some glycoproteins tested. The inhibition profiles with glycoproteins were different depending on hemagglutinin species, suggesting the presence of lectins specific for complex N-glycans, high mannose N-glycans or O-glycans. On the other hand, the activities of almost all algal hemagglutinins were stable over a wide range of pH and temperature, and independent of the presence of divalent cations, except Gelidiopsis scoparia hemagglutinin, its activity was dependent on the presence of divalent cations. These results suggest that Vietnamese marine macroalgae may be good sources of useful lectins for many biological applications.     

The biology and morphology of the marine harpacticoid copepod Heteropsyllus nunni Coull, during encystment diapause

Heteropsyllus nunni Coull, a meiobenthic harpacticoid copepod is the marine crustacean to undergo a state of diapause within a cyst. A 12 month field study indicated H. nunni adults reached peak population densities in winter, with nauplii maturing in the spring, becoming adults by April or May.At the last stage of development, a mature but unmated adult, they begin to prepare for encystment diapause. The copepods remain within their cyst in a state of diapause for 3–4 months during the summer only. Studies on the effects of temperature and photoperiod suggested that these two environmental cues are not crucial for induction or termination of diapause. Low temperature delayed development and time to encystment, while high temperatures accelerated development, making the time to encystment shorter. There were males than females in the cysts in laboratory experiments. Upon excystment, the copepods mate, and females begin egg production within one week. Adults that have excysted and mated die after a few weeks of active reproductive effort. Nauplii go on to mature and begin the univoltine diapause/reproductive cycle.The copepods prepare for dormancy in two ways: they begin to produce and store two types of secretory products to be used in cyst construction; then they produce large quantities of lipid to be used as a nutrient supply throughout diapause. Histochemistry of the cyst-building material indicated the lower urosome is full of two chemically different products. Dorsally, there is a storage sac of proteinaceous material. The ventral sac of secretory product is a mucopolysaccharide. The copepod builds the spherical cysts in a matrix of small and large sand grains. The cysts fit tightly around the ventral portion of the animal in the its flexed position: however, there is a large space between the cyst and the sides of the copepod.Biochemical analysis of the cyst showed it is composed of an amino acid complex similar to collagenous material. Scanning electron microscopy revealed a complex of large cuticular pores located in the lower urosome and caudal rami. There are specific pores for secretion of the two cyst-building products.     

A clarification of two congeners,Pseudodiaptomus lobipes and P. binghami (Calanoida,Pseudodiaptomidae) from India,with description of P. mixtus sp.n. from Bangladesh

Two closely related and often confused species of Pseudodiaptomus from the Lobus-species group, P. lobipes and P. binghami are redescribed from various locations along the east coast of India. These species predominately occur in freshwater though they can survive temporary periods of increased salinity. The distinctive features of the species are found on: female caudal ramal setae, female and male urosome 1–2 spinulation patterns, and fifth legs. A new species P. mixtus from Bangladesh is described.     

Diversity of the free-living marine and freshwater Copepoda (Crustacea) in Costa Rica: a review

The studies on marine copepods of Costa Rica started in the 1990’s and focused on the largest coastal-estuarine systems in the country, particularly along the Pacific coast. Diversity is widely variable among these systems: 40 species have been recorded in the Culebra Bay influenced by upwelling, northern Pacific coast, only 12 in the Gulf of Nicoya estuarine system, and 38 in Golfo Dulce, an anoxic basin in the southern Pacific coast of the country. Freshwater environments of Costa Rica are known to harbor a moderate diversity of continental copepods (25 species), which includes 6 calanoids, 17 cyclopoids and only two harpacticoids. Of the +100 freshwater species recorded in Central America, six are known only from Costa Rica, and one appears to be endemic to this country. The freshwater copepod fauna of Costa Rica is clearly the best known in Central America. Overall, six of the 10 orders of Copepoda are reported from Costa Rica. A previous summary by 2001 of the free-living copepod diversity in the country included 80 marine species (67 pelagic, 13 benthic). By 2009, the number of marine species increased to 209: 164 from the Pacific (49% of the copepod fauna from the Eastern Tropical Pacific) and 45 from the Caribbean coast (8% of species known from the Caribbean Basin). Both the Caribbean and Pacific species lists are growing. Additional collections of copepods at Cocos Island, an oceanic island 530 km away of the Pacific coast, have revealed many new records, including five new marine species from Costa Rica. Currently, the known diversity of marine copepods of Costa Rica is still in development and represents up to 52.6% of the total marine microcrustaceans recorded in the country. Future sampling and taxonomic efforts in the marine habitats should emphasize oceanic environments including deep waters but also littoral communities. Several Costa Rican records of freshwater copepods are likely to represent undescribed species. Also, the biogeographic relevance of the inland copepod fauna of Costa Rica requires more detailed surveys.     

Copepods act as a switch between alternative trophic cascades in marine pelagic food webs

A recent meta‐analysis indicates that trophic cascades (indirect effects of predators on plants via herbivores) are weak in marine plankton in striking contrast to freshwater plankton ( Shurin et al. 2002 , Ecol. Lett., 5, 785–791). Here we show that in a marine plankton community consisting of jellyfish, calanoid copepods and algae, jellyfish predation consistently reduced copepods but produced two distinct, opposite responses of algal biomass. Calanoid copepods act as a switch between alternative trophic cascades along food chains of different length and with counteracting effects on algal biomass. Copepods reduced large algae but simultaneously promoted small algae by feeding on ciliates. The net effect of jellyfish on total algal biomass was positive when large algae were initially abundant in the phytoplankton, negative when small algae were dominant, but zero when experiments were analysed in combination. In contrast to marine systems, major pathways of energy flow in Daphnia‐dominated freshwater systems are of similar chain length. Thus, differences in the length of alternative, parallel food chains may explain the apparent discrepancy in trophic cascade strength between freshwater and marine planktonic systems.     

Photophobic responses of calanoid copepods: possible adaptive value

A genual pattern of photophobic responses has been observedwhich differs for calanoid copepods from freshwater, estuarineand oceanic environments. Using a video-computer system formotion analysis, the photophobic responses of light and darkadapted calanoid copepods were compared. Dark-adapted copepodswere exposed to 600 ms flashes of dim blue light at 5 s intervalswhich simulated the flashes of biolumines-cent marine zooplankton.Light-adapted copepods were exposed to 600 ms intervals of darknessat 5 s intervals to simulate the shadows of organisms passingoverhead. Four species of coastal marine copepods (Acartia hudsonica,Centropages hamatus, Pseudocalanus minutus and Temora longicornis)all showed photophobic responses to both flashes and shadows.These responses may have adaptive value to the copepods sincethey live in an environment with predators that are bioluminescentat night and cast shadows on their prey during the day (e.g.ctenophores and cnidarian medusae). Two species of oceanic copepods(Euchaeta marina, Pleuromamma abdominalis) showed strong photophobicresponses to flashes but no response to shadows. This may correspondto the abundance of bioluminescent predators on copepods inthe oceanic environment (fish, ctenophores, siphonophores, etc.)and their lack of exposure to the shadows of predators, sinceboth these species are rarely found in the euphoric zone duringthe day. Two species of freshwater copepods (Diaptomus sanguineus,Epishwa massachusettsensis) showed no similar photophobic responseto flashes of light. This lack of startle response may relateto the lack of bioluminescence in the freshwater environment.Freshwater copepods showed a weak photophobic response to shadows.The adaptive value of this behavior is unclear, however, sincethe responses seem to be too weak to function for escape, andthe dominant predators large enough to cast shadows (fish) tendto approach their prey laterally. ¹Present address: Marine Science Institute, University of Texasat Austin, Port Aransas, TX 78373–1267, USA     

Two morpho-types of Botryllophilus (Cyclopoida,Ascidicolidae)

In taxonomic studies (in preparation) of the genus Botryllophilus from the eastern and western coastal waters of the Pacific and from the northern Gulf of California, we have found that the females are morphologically divided into 2 types. The 2 morpho-types in our material, which includes 10 species, are characterized by features of the urosome including anal segment and caudal rami, rostrum, antennule, antenna, mandible, maxillule, maxilla, maxilliped, 1st to 5th legs, and the apparatus at the oviducal aperture. These characters are described and the significance of the recognition of the 2 morpho-types is discussed for the purpose of solving taxonomic problems in Botryllophilus.     

Removal of Surface Glycoproteins and Transfer among <Emphasis Type="Italic">Brachionus</Emphasis> species

Glycoproteins on the body surface of females of the rotifer Brachionus plicatilis are a key signal in their mate recognition system. When B. plicatilis Russian strain females were exposed to 50 mM EDTA or EGTA, several surface glycoproteins were removed. Females exposed to EDTA died, but remained intact and were used in mating bioassays with conspecifics males. Live control females elicited a male mating response in 21% of encounters, freeze-killed control females elicited responses in 23%, but EDTA extracted females elicited a mating response in only 5% of encounters. At least some of the EDTA-extractable proteins on the surface of females appear to be critical to male mate recognition. EDTA treated females could be exposed to proteins extracted from other females and some proteins re-attached to their body surface, restoring their attractiveness to males. SDS-PAGE of these proteins revealed 15–17 prominent bands, most ranging in molecular mass from 66 to 12 kD. The EDTA-extractable proteins were separated using ion exchange chromatography and each fraction was tested for its ability to restore female attractiveness. When proteins in fraction 22 were bound to females, they restored 80% of the females’ ability to elicit male mating responses. Exposing EDTA treated females to bovine serum albumin or casein had no effect on their attractiveness to males. EDTA treated females from different Brachionus clades and species were exposed to proteins from fraction 22. Female attractiveness could be restored in most clades of B. plicatilis, but no transfer of mating attractiveness was observed to B. rotundiformis or B. ibericus females. Conspecific males treated with EDTA and exposed to proteins in fraction 22 could not be feminized and made attractive to other males. A sexual dimorphism in surface proteins therefore exists between B. plicatilis females and males. Successful transfer of glycoproteins critical in mate recognition is dependent on signal glycoprotein structure and the structure of the other proteins present on the surface of females.     

IDENTIFICATION OF MARINE DINOFLAGELLATES USING FLUORESCENT LECTINS1

Toxic and nontoxic species of marine dinoflagellates were characterized using fluorescent lectins. Lectin binding was detected by epifluorescence as well as spectrofluorometry. The binding assay of fluorescent lectins readily differentiated between morphologically similar species (i.e the toxic dinoflagellate Gymnodinium catenatum and the nontoxic Gymnodinium sp.). Lectins appear to be a useful tool to distinguish among different clones of the same species and, thus, possibly as a tool in dinoflagellate identification. Moreover, the lectins used show that thecate species have more binding sites and diversity in glycan moieties than athecate species.     

Toxicity of blooms of the cyanobacterium Trichodesmium to zooplankton

The marine filamentous bloom-forming cyanobacteria Trichodesmium thiebautii and T. erythraeum were collected at locations in the Carribean during Jan.–Feb. 1991. They were screened for toxicity using Artemia salina and several species of copepods, which were harpacticoid grazers, filter-feeding calanoids, or cyclopoid copepods. Approximately 50% of the 89 T. thiebautii samples caused> 50% lethality of A. salina, though none of the 16 T. erythraeum samples caused> 25% lethality. The T. thiebautii bloom samples were toxic to the calanoid and cyclopoid copepods and non-toxic to the harpacticoid grazers. In contrast the T. erythraeum bloom samples were not toxic to any copepods tested.     

Egg-brooding, body size and predation risk in planktonic marine copepods

We investigated the interacting effects of copepod body size and the presence or absence of egg masses on the risk of predation by a visual predator. We conducted selection experiments involving three-spined sticklebacks (Gasterosteus aculeatus) and copepods ranging in body mass from 0.5 to 740 μg C: Oithona similis, Corycaeus anglicus, Pseudocalanus newmani, P. moultoni, Pseudodiaptomus marinus, and Paraeuchaeta elongata. We found that sticklebacks selected ovigerous females of the two smallest-bodied species of copepods (Oithona similis and Corycaeus anglicus). In contrast, the fish showed no significant selection for ovigerous females of the remaining, larger-bodied species. Unexpectedly, egg mass position (i.e., in a ventral, dorsal or lateral location on the urosome) appeared to influence predation risk more than did body size, resulting in higher predation risk for the cyclopoid and poecilostomatoid species than for the calanoid species we tested. Although the sticklebacks showed no statistically significant preference for ovigerous females of any of the four calanoid species, for each species the overall proportion of ovigerous females ingested was slightly greater than 0.50. Thus, whether body size influences the susceptibility of egg-brooding calanoid copepods to predation remains an open question. Received: 24 August 1998 / Accepted: 6 July 1999     

Freshwater components in the diet of the marine neustonic fish,Urophycis tenuis (Mitchill)

Synopsis Stomach content analysis of a neustonic marine juvenile white hake (Urophycis tenuis (Mitchill)), revealed the presence of a unusual 0.70 mm bolus, consisting of a freshwater rotifer, Keratella cochlearis, and several species of freshwater phytoplankton: Asterionella formosa, Melosira granulata, Dinobryon bavaricum and Dinobryon cylindricum. Other gut items were typical for the predator during the August period, consisting of various stages of marine calanoid copepods and cladocerans; average prey length was 0.74 mm. The possibility of bolus introduction as a secondary diet component was eliminated; evidence indicated that the particle was ingested in toto. Consideration of a number of alternative mechanisms for bolus acquisition led to the conclusion that the bolus was disseminated via the ballast tank or contaminated freshwater holding tank of a ship. Urophycis tenuis appears to feed selectively on the basis of particle size. Consequently, this type of neustonic predator may be particularly vulnerable to man-made pollution and oil spills.     

Assessing the diversity of New Zealand freshwater harpacticoid copepods (Crustacea: Copepoda) using mitochondrial DNA (COI) barcodes

Taxonomic and ecological studies of freshwater harpacticoid copepods are limited globally by the ability to easily and accurately identify specimens. Here, we test the use of the mitochondrial cytochrome c oxidase subunit I (COI) gene locus as a tool for assessing the diversity of freshwater Harpacticoida. We obtained sequences from New Zealand harpacticoid copepods, representing two families, five genera and nine species, including the non-indigenous Elaphoidella sewelli. All species were delineated by the COI gene. However, high intraspecific diversity was evident among populations of Elaphoidella bidens (>12%), and between North and South Island populations of Bryocamptus pygmaeus (>18%), potentially indicating the presence of morphologically cryptic taxa. We suggest that mitochondrial DNA (COI) sequences can provide a useful tool for the routine identification of freshwater harpacticoid copepods. Applications of these data will include assessing species diversity and biogeography as well as assisting with the detection of non-indigenous species.     

<Emphasis Type="Italic">Prorocentrum rivalis</Emphasis> sp. nov. (Dinophyceae) and its phylogenetic affinities inferred from analysis of a mixed morphological and LSU rRNA data set

A new freshwater epiphytic Prorocentrum species, Prorocentrum rivalis, from the temperate region of the Haute-Vienne, France, is described. This species is the third freshwater species identified among approximately 60 marine Prorocentrum species. This new species is described using scanning electron microscope and phylogenetic analyses by a polyphasic approach (LSU rRNA sequences combined with 9 morphological characters). The phylogenetic analysis attests that P. rivalis is close to other planktonic freshwater species and the freshwater Prorocentrum clade is evolutionarily derived from an epiphytic freshwater prorocentroid ancestor. The unique marine species in the freshwater clade results from an ecophysiological reversion. P. rivalis differs from other epiphytic taxa by its rarity, its temperate distribution and its ecophysiological needs. The phylogeny confirms also that all planktonic Prorocentrum species are evolutionarily derived from epiphytic/benthic ancestors.     

Historical biogeography and distribution of the freshwater calanoid copepods (Crustacea: Copepoda) of the Yucatan Peninsula, Mexico

Aim To determine and analyse the distribution of the freshwater calanoid copepod (Diaptomidae and Pseudodiaptomidae) fauna of the Yucatan Peninsula (YP) and its relation to the geological history of this Neotropical karstic plain. Location The Yucatan Peninsula, Mexico. Methods Plotting of geo‐referenced sites, analysis of local and regional geological history, analysis and comparison of regional and local records. Results The current composition and distribution of Diaptomidae and Pseudodiaptomidae in the YP mainly reflects recent, post‐Pliocene colonization events. This invasion did not reach America, but only parts of Central America (CA). The presence of diaptomids in the continent since the pre‐Cretaceous and the presumed post‐Cretaceous (Palaeocene–Oligocene) radiation of Diaptomidae in Middle America suggest earlier colonizations of the YP. The marine transgressions kept most of the YP submerged in different geological periods, thus eliminating any original primary freshwater colonizers, such as the diaptomids. The periods of marine regression probably represented opportunities for new waves of diaptomid colonization of the YP. The latest dispersal of diaptomids in the YP during the Holocene (8000 yr bp ) was probably an intermittent process because of the alternative dry and wet periods and interglacial transgressions. The presence of the Nearctic Leptodiaptomus and Arctodiaptomus in the YP and the current distribution of Mastigodiaptomus might represent remnants of earlier invasions of Diaptomidae in Middle America. The Neotropical Mastigodiaptomus probably originated in the Late Cretacic CA/proto‐Antilles complex. Forms derived from a M. albuquerquensis type ancestor radiated into the YP leaving relatively isolated populations of three species in the northern half of the peninsula. The distribution of the brackish water Pseudodiaptomus marshi well inside the coastline might have resulted from stranding and subsequent adaptation of this species during a marine regression in the Bacalar formation; this agrees with the vision of this taxon as being in process of invasion of freshwater environments. Main conclusions It is not probable that the South American (SA) diaptomid fauna originated from an invasion of upper Neotropical/Nearctic forms. The current distribution of freshwater calanoid copepods reflects relatively recent, post‐Pliocene biogeographical patterns, but probably older patterns are involved as well. The northern and eastern coasts of the Yucatan are the most recently colonized by diaptomids. Differing from other freshwater groups surveyed in the Yucatan that have marine relatives (i.e. fishes, amphipods, isopods, mysids, macrocrustaceans), there is no evidence of local vicariant events involving cave‐dwelling forms or marine relicts in the diaptomid fauna of the YP.     

Evidence of prezygotic isolation,but not assortative mating,between locally adapted populations of Fundulus heteroclitus across a salinity gradient

Selection along environmental gradients can drive reproductive isolation and speciation. Among fishes, salinity is a major factor limiting species distributions, and despite its importance in generating species diversity, speciation events between marine and freshwater are rare. Here, we tested for mechanisms of reproductive isolation between locally adapted freshwater and brackish water-native populations of killifish, Fundulus heteroclitus, from either side of a hybrid zone along a salinity gradient. There was evidence for pre-zygotic endogenous reproductive isolation with reduced fertilization success between crosses of freshwater-native males and brackish water-native females. Exogenous pre-zygotic isolation was also present where females had highest fertilization in their native salinity. We used a replicated mass spawning design to test for mate choice in both brackish and fresh water. After genotyping 187 parents and 2523 offspring at 2347 SNPs across the genome, 85% of offspring were successfully assign to their parents. However, no reinforcing mate choice was observed. These results therefore demonstrate emerging, yet limited, reproductive isolation and incipient speciation across a marine to freshwater salinity gradient and suggest that both endogenous and exogenous mechanisms, but not assortative mating, contribute to divergence.