Microsatellites from the vairone Leuciscus souffia (Pisces: Cyprinidae) and their application to closely related species

Eleven microsatellites were isolated from the vairone Leuciscus souffia (Risso 1826), an endangered fish that inhabits river systems in and around the Alps in Europe. The level of genetic diversity was assessed in 29 individuals of the subspecies L. s. souffia, and their variability was further estimated in seven individuals of a different subspecies, L. s. muticellus. Eight of these microsatellite loci were also applied to seven closely related cyprinid species. Availability of the reported microsatellite loci will facilitate the investigation of population genetic structure of these species with applications for the development of conservation strategies and phylogeographical approaches.     

Evergreen Foliage Contributions to the Spring Growth of Taxus

Taxus media cv. Hicksii plants were grown one season under a low and high level of nitrogen fertilization. Before growth in the spring the plants were divided into two groups, one of which was defoliated and the other left intact. The growth and spring utilization of the nitrogen and carbohydrate reserves of defoliated plants were compared to the intact plants 0, 2, 4 and 6 weeks after growth started in the spring. The plants were separated into buds (all new growth), roots and stems and analyzed for changes in total nitrogen, basic and non-basic amino acids, hemicelluloses, soluble sugars, organic acids and chlorophyll. The older evergreen needles from plants grown under low nitrogen levels contain 20 % of the carbohydrate and 24% of the nitrogen used in spring growth. The needles from plants grown under high nitrogen levels contained 56% of the carbohydrate and 49% of the nitrogen used in spring growth. Removal of the old needles before spring growth removed this nitrogen and carbohydrate reserve and reduced the total plant chlorophyll content after 6 weeks of growth to 50% of that found in intact plants, with the result that defoliated plants did not show a growth response to nitrogen. Amino acids accumulated in the stems and buds of defoliated plants as carbohydrates became limiting. The defoliated plants removed 25% more available carbohydrates from the roots and stems than intact plants and their buds contained 50% less available carbohydrates. Plants without old needles showed similar growth rates under low and high nitrogen regimes and produced 33% of the dry weight of intact plants grown under high nitrogen levels and 66% of the dry weight of intact plants grown under low nitrogen levels. The old needles of taxus plants contain substantial amounts of reserve nitrogen and carbohydrate and these needles greatly influence the extent and rapidness of growth in the spring. When the needles are removed, the other tissues can supply an adequate amount of nitrogen but the carbohydrate supply becomes limiting for spring growth.     

Ecological and evolutionary consequences of the trophic polymorphism in Cichlasoma citrinellum (Pisces: Cichlidae)

The neotropical cichlid fish Cichlasoma citrinellum is polymorphic in the structure of its pharyngeal jaw apparatus and external morphology. The pharyngeal jaws are either gracile and bear slender, pointed teeth (papilliform) or robust with strong, rounded teeth (molariform). Molariform morphs have a ‘benthic’, and papilliform morphs a ‘limnetic’ body form. Furthermore, this species is also polychromatic, with yellow and black morphs. The molariform morphology of the pharyngeal jaw apparatus adapts the fish for cracking and feeding on snails. Based on analysis of stomach contents, 94% of the molariform morph ate snails whereas only 19%, of the papilliform morph did so. This result suggests that the morphs occupy different ecological niches. The morphology of the pharyngeal jaw apparatus does not correlate significantly with sex, but it does with body colouration (P<0.005). Cichlasoma citrinellum mate assortatively with their own colour; therefore a mating preference for colour may lead to genetic isolation of trophic morphs. The frequency of the molariform morph differs strikingly among populations of five Nicaraguan lakes and its abundance is correlated with the abundance of snails, the fishes' principal prey item. Among populations the frequency of molariform morphs decreases in the dry season. Morphology possibly changes reversibly within particular individuals between seasons. These results suggest that phenotypic plasticity and polymorphisms may be an adaptive characteristic of cichlid fishes. Patterns of intraspecific morphological variation match patterns of interspecific morphological diversification which suggests that universal developmental mechanisms canalize the possible expressions of morphology. The ability to respond morphologically to environmental shifts, in conjunction with genetically determined trophic polymorphisms and sexual selection via mate choice, could be the basis for speciation through intermediate stages of polymorphism of the impressive adaptive radiation of cichlid fishes.     

Preservation of in situ reef framework in regions of low hurricane frequency: Pleistocene of Curaçao and Bonaire, southern Caribbean

Luxuriant fringing reefs along the southwestern shores of the Caribbean islands of Curaçao and Bonaire (12°N), located outside the most frequent hurricane tracks, are rarely affected by major storms. Consequently, reef growth and long-term preservation are potentially optimal and distinct from reefs experiencing greater hurricane frequency. Hurricane Lenny (November 1999) took an unusual west-to-east track, bisecting the Caribbean Basin north of these islands, but generated heavy waves (3-6  m) that severely damaged reefs along the normally leeward shores. Massive coral colonies >100 years old were toppled, but even at the most severely damaged sites, 82–85% of colonies remained in growth position. Late Pleistocene (125  ka) elevated reefs in the Lower Terrace of Curaçao record even higher proportions of corals in growth position (93%), possibly reflecting a low hurricane frequency during the Pleistocene highstand. In comparison, coeval Pleistocene reefs in regions that today experience a high hurricane frequency (Great Inagua Island and San Salvador, Bahamas) have lower proportions of corals preserved in growth position (79% and 38%, respectively). These results are consistent with the hypothesis that reefs in regions experiencing very low hurricane frequency, like the southern Caribbean, are more likely to be preserved with corals in primary growth position in comparison to regions with higher hurricane frequency.     

In Vitro Excystation of Spironucleus muris

ABSTRACT. In vitro excystation of Spironucleus muris cysts, purified by sequential sucrose and Percoll gradients from mouse feces, was studied. Three in vitro excystation procedures, used for Giardia , were assessed to determine the most useful method. Excystation was monitored by light microscopy and subsequently characterized by transmission and scanning electron microscopy. Spironucleus muris excysted routinely at a level greater than 90% when induced in Hanks' balanced salt solution containing sodium bicarbonate at pH 2.0 and transferred to Tyrodes' salt solution as an excystation medium. Similarly, high rates of excystation were recorded after induction of S. muris cysts in 0.1 M potassium phosphate buffer (pH 7.0) with sodium bicarbonate and excystation in trypticase-yeast extract-iron medium (TYI medium) or phosphate-buffered saline. A lower rate and percentage of excystation were observed after induction of S. muris cysts in an aqueous hydrochloric acid solution (pH 2.0) followed by excystation in TYI medium. All excystation methods produced extremely active S. muris trophozoites with normal morphology. Nonexcysting S. muris cysts have a wall composed of an outer fibrous and an inner membranous portion. Following induction, numerous vesicles appeared in the peritrophic space. Excystation began by the cyst wall opening at one pole, and the anterior part of the trophozoite protruding from the cyst wall. The trophozoite emerged progressively from the cyst wall and the empty cyst wall appeared to collapse. Excysted trophozoites exhibited normal morphological features of S. muris trophozoites isolated from the mouse intestine.