Morphological and biochemical variation in populations of Dugesia (G.) tigrina (Turbellaria, Tricladida, Paludicola) from the western Mediterranean: biogeographical and taxonomical implications

Dugesia (Girardia) tigrina is a Nearctic freshwater triclad native to North America, now amply distributed in most European countries as the result of a reportedly unique introduction into north-west Europe early this century and its dispersal southwards and eastwards in the last decades. All European populations described so far are externally spotted, with a pigmented pharynx, and, apart from two sexual populations recently described in England, they are fissiparous. This uniformity contrasts with the variability reported in its native area, thus supporting the hypothesis of a single introduction. However, thorough morphological, karyological and biochemical comparative studies of European populations have never been properly done.
We have undertaken such a study comparing 24 populations present in a restricted area of the western Mediterranean. The results show an unexpectedly ample variation in external and pharyngeal pigmentation patterns, the presence of sexually reproducing populations, and ample variation in their biochemical patterns as revealed by isoelectric-focusing. This is an indication that European Dugesia (G.) tigrina is far more diverse than previously thought, suggesting that several, independent introductions from its native area have recently occurred, and the implications of these findings in biogeographical and taxonomic terms are discussed.     

Measured and Calculated Transpiration in Trifolium repens under Different Water Potentials

The relationship between transpiration measured gravimetrically,a generalized transpiration equation, and the ratio VPD/r_leafwas investigated in Trifolium repens plants subjected to varyingwater potentials. Dawn leaf water potential was measured witha pressure chamber, leaf diffusion resistance with a diffusionporometer, leaf temperatures with a thermistor, and relativehumidity with an aspirated psychrometer. During drought transpirationrates determined by both methods were quite similar particularlyat the lowest water potentials. After rewatering calculatedrates were somewhat higher than measured ones. It is concludedthat transpiration calculated by the indirect method is a usefuland reasonable estimate of transpiration for single plants undervarying water potentials.     

Comparative Study of Experimental Ocular Melanoma Using Two Implantation Techniques of B16-F10 Melanocytes

Objective: The aim of this work was to evaluate the behaviour of B16-F10 melanoma cell cultures implanted in the anterior chamber of the eye of New Zealand white rabbits by studying the clinical-pathological and ultrastructural characteristics of the lesions. Methods: One group (A) (consisting of 30 rabbits) was transclerally inoculated (1 mm from sclero-corneal limbus) with 4×10⁶ melanocytes and another group (B) (also 30 animals) was inoculated once per week for 3 consecutive weeks with 5×10⁶ cells (total 15×10⁶); 30 animals acted as the control group (C). All the lesions were processed for optic and electronic microscopy. Results: Tumoral growth in group A was 43% (13/30) and in group B 80% (24/30). All lesions were pigmented and none perforated the eyeball. Microscopically, they were a mixture of epithelioid and fusiform cells disposed around the blood vessels. Ultrastructurally, the presence of melanosomes in different stages of maturation and aberrant melanosomes were characteristic. Conclusion: We suggest that the transcleral inoculation of 15×10⁶ B16-F10 melanocytes into the anterior chamber of the eye of New Zealand white rabbits may be a valid and reproducible method for obtaining an experimental ocular melanoma model.     

Myxosoma cerebralis (Myxozoa: Myxosporea) Etiologic Agent of Salmonid Whirling Disease Requires Tubificid Worm (Annelida: Oligochaeta) in its Life Cycle

Studies of the life cycle of Myxosoma cerebralis showed that development of infectivity did not occur endogenously but that the spore “aging” process required participation of an aquatic tubificid oligochaete. Data suggestive of such involvement were derived from trials in which spores were “aged” in an array of inert, sterilized, pasteurized, or natural aquatic substrates and from examination of aquatic soils from trout hatcheries in which whirling disease was epizootic. The role of the aquatic oligochaete was confirmed two ways. First, signs of whirling disease developed, and M. cerebralis spores were produced in young rainbow trout (Salmo gairdneri) that had been fed oligochaetes harvested from pond soil taken from two hatcheries where whirling disease was epizootic. Second, when containers of pasteurized soil were populated with four genera of oligochaetes–Aeolosoma, Dero, Stylaria, or Tubifex– from a biological supply house, or with tubificid worms from trout hatcheries free of whirling disease, and then seeded with M. cerebralis spores and “aged” for 4 months, whirling disease occurred only in trout held with Tubifex and with hatchery tubificids.