Tardigrades from Portugal: four new records and description of two new species
PAULO FONTOURA1, GIOVANNI PILATO2, OSCAR LISI2 & PAULO MORAIS1
1Eco-Ethology Research Unit (FCT-331/94) and Department of Zoology and Anthropology, Faculty of Sciences, University of Porto, R. Campo Alegre s/n, FC4, 4169-007 Porto, Portugal. E-mail: [email protected] di Biologia Animale “Marcello La Greca”, Università di Catania Via Androne 81, 95124 Catania, Italy. E-mail: [email protected]
Abstract
Six species of Eutardigrada are recorded from Portugal; four of them, Macrobiotus crenulatus Richters, 1904, Hypsibius seychellensis Pilato, Binda & Lisi, 2006, Diphascon (Diphascon) pingue (Marcus, 1936) and D. (Diphascon) pataneiBinda & Pilato, 1971 are recorded for the first time in Portugal. Two species, Minibiotus orthofasciatus sp. nov. and Bertolanius (new name of Amphibolus) portucalensis sp. nov. are new to science. Minibiotus orthofasciatus sp. nov. is one of the species of the genus with three macroplacoids, microplacoid and cuticular pores forming transverse bands. The new species differs from all existing species by one or more of the following characters: distribution of the pores, shape of the pores, absence of dots on the legs, level of insertion of the stylet supports on the buccal tube. To the new species is attributed an unembryonated egg similar to those of Minibiotus intermedius (Plate, 1888), M. poricinctus Claxton, 1998, M. floriparus Claxton, 1998, and M. weglarskae Michalczyk, Kaczmarek & Claxton, 2005 but different from them in some details. Bertolanius portucalensis sp. nov. is very similar to the other species of the genus, but it differs from them in having very small cuticular tubercles. From some of them it differs by characters of the bucco-pharyngeal apparatus and/or of the eggs. This is the first record of the genus and of the Eohypsibiidae family in the Iberian Peninsula.
Since the first note about Portuguese tardigrades in 1937, describing three species (Barros-e-Cunha, 1937) only a few studies dealing with this group of micrometazoans have been carried out in Portugal (excluding the archipelagos of Madeira and Azores). An important contribution (seven papers reporting 31 records, including the description of three new species) was made during the subsequent ten years by Da Cunha (1941, 1943,
1944a,b, 1947a,b, 1948). After a gap of about 20 years a new short phase of study, unfortunately without continuity, occurred with contributions by Fontoura (1981, 1982), Maucci (1983) and Maucci & Durante-Pasa
(1984a,b,c). As a result of these studies, a final number of 57 limno-terrestrial tardigrade species have been identified in Portugal, including the descriptions of 8 species new to science. After the proposal of a new systematic arrangement of the families and genera of the Eutardigrada (Pilato 1969), in parallel with the improvement of the technical means of analysis, the taxonomy of the group underwent considerable changes. Very probably there are some mis-identifications and the number of 57 Portuguese species could be reduced
even more, as proposed by Maucci & Durante-Pasa (1984c). However, the opinion of these authors was based only on the observation of the published figures which, from our point of view, are very weak arguments. The corrections of the synonymies and the mistakes detected in the previous records are accepted because it is probably impossible to re-examine the studied material (in its majority probably lost). We prefer to respect, with caution, the outcome achieved in the literature and to wait until future research can solve the existing
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inaccuracies. Taking in mind the current biodiversity crisis and consequently the need to create a legacy of taxonomic knowledge for the future (Wheeler et al., 2004; Carvalho et al., 2005) and the very poor knowledge of the Portuguese fauna, we decided to reactivate studies on this subject.
In this paper the first record in Portugal of four species of limno-terrestrial tardigrades is reported (Macrobiotus crenulatus Richters, 1904, Hypsibius seychellensis Pilato, Binda & Lisi, 2006, Diphascon (Diphascon) pingue (Marcus, 1936) and D. (D.) patanei Binda & Pilato, 1971) as well as the description of two species new to science: Minibiotus orthofasciatus and Bertolanius portucalensis. Bertolanius is the new name attributed to Amphibolus by Özdikmen (2008). This is the first record of this genus and of the Eohypsibiidae family in the Iberian Peninsula.
After this study 63 limno-terrestrial tardigrade species are recorded for Portugal.
Material and methods
Tardigrades were extracted from dried lichen and moss samples collected in Northern Portugal: Amarante (41º16’24’’N 8º03’48’’W) in November 2005 and February 2006 and Avintes (Parque Biológico de Gaia, 41º05’57’’N 8º33’41’’W) in May 2006 and April 2008. The great majority of the specimens were mounted in Hoyer’s medium with a small amount of potassium iodide solution (Horning et al., 1978), only a few specimens were mounted in polyvinyl lactophenol. Measurements, given in micrometers (µ m), and photomicrographs were made under oil immersion, using a Zeiss Phase Contrast Microscope equipped with digital camera and using Axiovision 4.6.3 Imaging System Software. Structures were measured only if they were undamaged and their orientation was suitable. Body length was measured from the anterior margin to the end of the body, excluding the hind legs. Measurements of the bucco-pharyngeal apparatus and of the claws, and the assessment of the pt index, were obtained according to Pilato (1981).
For comparison, we examined the neotype of Macrobiotus crenulatus; holotype of Minibiotus sidereusPilato, Binda & Lisi, 2003; paratypes of Minibiotus bisoctus (Horning, Schuster & Grigarick, 1978), Minibiotus poricinctus Claxton, 1998; Minibiotus floriparus Claxton, 1998; Minibiotus ethelae Claxton, 1998; Minibiotus keppelensis Claxton, 1998; Minibiotus gumersindoi Guil & Guidetti, 2005; specimens of Minibiotus intermedius (Plate, 1888) and Minibiotus furcatus (Ehrenberg, 1859); paratypes of Bertolaniusweglarskae (Dastych, 1972) and Bertolanius volubilis (Durante-Pasa & Maucci, 1975); specimens of Bertolanius smreczynskii (Węglarska, 1970); the holotype of Hypsibius seychellensis; the holotype of Diphascon patanei and specimens of Diphascon pingue.
The paratypes of Minibiotus bisoctus were on loan from the Museum of New Zealand (Wellington); paratypes of Minibiotus gumersindoi by the “Museu Nacional de Ciencias Naturales de Madrid” (Spain); the rest of the above mentioned material is deposited in the collection of Binda & Pilato (Museum of the Department of Animal Biology, University of Catania).
Results
Macrobiotus crenulatus Richters, 1904
Material examined: Three specimens (body length 229.3–280.4 µm) collected from a moss sample at Amarante were mounted in Hoyer’s medium. The species, described by Richters in 1904 and considered as subspecies of M. echinogenitus by the same author in 1911, has been considered bona species by Binda (1988).
Distribution: Species widely distributed in Europe and North America.
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Minibiotus orthofasciatus sp. nov. Figs. 1, 2
Material examined: Thirty specimens and one unembryonated egg, mounted in Hoyer’s medium, collected on lichens from tree trunks at Parque Biológico, Avintes – Vila Nova de Gaia (six specimens and the egg were collected in May 2006, all the other specimens were collected in April, 2008). Four specimens were in simplex stage and 1 in molting stage.
Type repository: The holotype (slide N. 5231), a paratype (slide N. 5232) and the egg (slide N. 5233) are deposited in the collection of Binda and Pilato (Museo del Dipartimento di Biologia Animale “Marcello La Greca”, Università di Catania); the other paratypes (slides CI-1 to CI-17) in the collection of P. Fontoura (Department of Zoology and Anthropology, Faculty of Sciences, University of Porto).
Specific diagnosis: Cuticle with clearly defined eleven dorsal and eight ventral transverse bands of pores various in shape and size. Pores larger on cephalic and caudal portions of the body. Dots on the legs absent but very fine, almost invisible dots, seem to be present on all the dorsal surface of the body. Three macroplacoids (the first is the largest, second and third very similar in length) and a very minute microplacoid present. Claws with developed accessory points. Smooth lunules present.
FIGURE 1. A–D, Minibiotus orthofasciatus sp. nov.: A, anterior portion of the body of a paratype, the dorsal bands of pores are visible; B, postero-dorsal bands of pores of a paratype; C, details of second and third leg pores of a paratype; D,
posterior portion of the body of a paratype, the ventral bands of pores and the claws are visible (scale bars = 10 µm).
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Description of the holotype: Body length 186 µm, colorless, eye spots present, visible on living specimens and before slide mounting. Cuticle smooth with pores of varying size (diameter 1.0 to 2.1 µm) and shape: roundish, oval, trilobed and, rarely, quadrilobed. The pores are arranged in dorsal and ventral bands constant in number and almost constant in pore distribution (Figs. 1A, B, C, D).
Dorsally eleven bands are present (Figs. 1A, B): two cephalic ones formed by two transverse rows; three bands also formed by two rows at the level of the first three pairs of legs; three bands, single on the dorsal wall but double on the lateral walls, are present after each pair of legs; and three caudal single bands (near the single bands very rarely supplementary pores may be present). On the ventral surface of the body eight distinct bands of a few roundish pores are present (Fig. 1D). A few pores are also present on the legs. No granulation is visible on the legs. Extremely fine, almost invisible, dots are probably present on the dorsal surface of the body, but the presence of this granulation needs to be confirmed.
Mouth antero-ventral. Buccal armature not visible with light microscope. Rigid buccal tube 22.6 µm long, with an anterior bend (Fig. 2B); a posterior bend is almost absent; a thickening of the buccal tube wall is present caudal to the stylet supports where the buccal tube is 1.8 µm wide externally (pt = 8.0). Buccal tube with ventral lamina not particularly short (pt = 58.4 measured in lateral view) (Fig. 2B). Stylet supports inserted on the buccal tube at 66.8% of its length (pt = 66.8). Pharyngeal bulb with large triangular apophyses, three small macroplacoids (Figs. 2 A, B) and a very small microplacoid (Fig. 2B); first macroplacoid, pear shaped, and partially lying under the apophyses as in other species of the genus, 3.1 µm long (pt = 13.7); second macroplacoid 2.4 µm long (pt = 10.6); third macroplacoid 2.4 µm long (pt = 10.6). The very small microplacoid, 0.9 µm long (pt = 4.0), is difficult to see. Macroplacoid row length, excluding the microplacoid, 7.4 µm long (pt = 32.7); entire row of placoids 8.3 µm long (pt = 36.7).
Claws of hufelandi type, well developed (Figs. 1C, D). Primary branches of internal and external claws with well developed accessory points. External and internal claws of legs II and III 6.8 µm long (pt = 30.1) and 6.3 µm long (pt = 27.9) respectively, including accessory points. Posterior and anterior claws of fourth pair of legs 6.9 µm long (pt = 30.5) and 6.8 µm long (pt = 30.1) respectively.
Smooth lunules, better developed on the fourth pair of legs (2.5 µm longer diameter), are present. A small cuticular bar is present near the lunules on the first three pairs of legs.
The measurements of some structures of holotype and three paratypes are shown in Table 1.One unembryonated egg has been found with the described specimens. It is an egg very similar to those of
some other species of Minibiotus (M. intermedius, M. floriparus and M. poricinctus) with screw-like processes each surrounded by a membranous hood (Figs. 2C, D, E); therefore we think that it may be attributed to the only species of Minibiotus genus found in the sample (other species collected in the same sample were Milnesium tardigradum Doyère, 1840; Diphascon pingue; Macrobiotus hufelandi Schultze, 1834; Macrobiotus recens Cuénot, 1932 of which the eggs were also present).
The diameter is 54.0 µm including the processes, 48.0 µm excluding them; the processes (Fig. 2C) are 4.2–4.5 µm in height with a base diameter of 2.3–2.7 µm and a top diameter of 3.3–3.6 µm. The processes are 6.4–6.9 µm apart. About 24 processes are present around the circumference and about 60 in the hemisphere. The base of the membranous hood of each process is connected to the surrounding ones and, as a consequence, the egg shell appears areolated (Figs. 2D, E); generally six polygonal areolae are present around each process, and their margins appear formed by many dots (Fig. 2E).
Etymology: The name orthofasciatus refers to the very regular straight (ortho) disposition of the bands (fasciatus) of cuticular pores.
Differential diagnosis: The presence of very fine cuticular dots has to be confirmed; therefore this character is not taken into consideration in the following comparisons.
The distribution of the cuticular pores in Minibiotus orthofasciatus sp. nov. is almost identical to that of Macrobiotus striatus Mihelčič, 1949, which probably has to be transferred into the genus Minibiotus as hypothesized also by Guidetti et al. (2007), but it differs from this species in many characters: colorless (M. striatus is yellow), eye spots present, cuticular pores of various size and shape, first macroplacoid longer than the third, a small microplacoid present, different egg.
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TABLE 1. Measurements (in µm) and pt values of some structures of the holotype, and three paratypes, the smallest and largest ones included, of Minibiotus orthofasciatus sp. nov.
The distribution of the cuticular pores of the new species is also similar to that of Minibiotus pustulatus(Ramazzotti, 1959) but the new species differs from this species in having smaller caudal pores (in M. pustulatus the diameter is 6–7 µm), eye spots present and different egg.
Nine other species of Minibiotus have three macroplacoids and cuticular pores arranged in distinct bands. One of them, M. gumersindoi, has pores with a distribution similar to that of Minibiotus orthofasciatus sp. nov., but the new species differs from it in also having trilobed and quadrilobed pores, the stylet supports inserted on the buccal tube in a more caudal position (pt = about 67 in the new species, 50–57.5, according to Guil & Guidetti (2005) and confirmed by us, in M. gumersindoi), longer ventral lamina, slightly larger macroplacoids and smaller microplacoid.
The new species differs from the remaining eight species with three macroplacoids and microplacoid in having a different pore distribution. In addition it differs from M. poricinctus and M. keppelensis in also having trilobed and quadrilobed pores, in lacking granulation on the legs, in having the stylet supports inserted on the buccal tube in a more caudal position (pt = about 67 in Minibiotus orthofasciatus sp. nov., about 60 in M. poricinctus and about 61 in M. keppelensis) and in having longer ventral lamina (pt = about 58in Minibiotus orthofasciatus sp. nov., about 36 in M. poricinctus and about 40 in M. keppelesis).
smallest paratype holotype paratype largest paratype
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Minibiotus orthofasciatus sp. nov. differs from M. bisoctus and M. weglarskae in lacking granulation on the legs and in having the stylet supports inserted on the buccal tube in a more caudal position (pt = about 67in the new species, about 60 in M. bisoctus and 54.5–59.6 in M. weglarskae) and in having longer ventral lamina (pt = about 58 in M. orthofasciatus, about 42 in M. bisoctus); in the description of M. weglarskae the authors did not indicate the length of the ventral lamina, but they wrote that it is “very short”, and their figures 7, 14, and 15 show that this structure is clearly shorter than in M. orthofasciatus sp. nov. In addition M. orthofasciatus sp. nov. differs from M. bisoctus in having eye spots, and from M. weglarskae in having a small cuticular thickened bar near the lunules on the first three pairs of legs.
FIGURE 2. A–E, Minibiotus orthofasciatus sp. nov.: A, bucco-pharyngeal apparatus of a paratype, the thickening of the buccal tube wall is shown; B, bucco-pharyngeal apparatus of the holotype; C, egg, arrows indicates the egg processes; D, areolated design of the egg shell formed by the connections of the membranous hood of the processes (arrows); E, polygonal areolae around processes with dots on their margins are shown (scale bars = 10 µm).
The new species differs from Minibiotus furcatus and Minibiotus eichorni Michalczyk & Kaczmarek, 2004, in lacking granulation on the legs and in having a smaller microplacoid. From M. furcatus also in having narrower buccal tube (unfortunately, due to a qui pro quo, Binda and Pilato, in a paper of 1992, wrote that the pt value relative to the buccal tube in M. furcatus is 7.94; we noted the mistake, and we found a value 11.2).
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Minibiotus orthofasciatus sp. nov. differs from Minibiotus granatai (Pardi, 1941) and M. ethelae in having the cuticle of the caudal portion not different (not more thickened) from that of other parts of the body, and without gibbosities. It also differs from M. ethelae, M. furcatus, M. keppelensis and M. weglarskae in having a different egg. The eggs of M. bisoctus, M. gumersindoi and M. eichorni are not known. The egg found with the new species and, according to us attributable to it, is similar to that of M. intermedius, M. floriparus, M. weglarskae and M. poricinctus but it differs from them in some details. It differs from the egg of M. intermedius because it has processes larger and less numerous (Table 2); in addition the egg shell is areolated and small dots are present where the membranous hoods reach the egg shell. Also the egg of M. poricinctus has dots at the base of the membranous hoods, but the egg attributed to the new species has smaller and more numerous processes (Table 2). The egg attributed to Minibiotus orthofasciatus sp. nov.differs from those of M. floriparus in having areolated egg shell with dots where the membranous hoods reach the egg shell; in addition it is smaller, with smaller processes and, in particular, with smaller terminal portion (Table 2) where a ring of pores well evident in M. floriparus, is almost invisible. The egg attributed to the new species differs from those of M. weglarskae in having areolated egg shell with dots where the membranous hoods reach the egg shell; in addition it has smaller processes with smaller terminal portion (Table 2) where a ring of pores, well evident in M. weglarskae, is almost invisible.
TABLE 2. Measurements (in µm) and other characteristics of the egg of Minibiotus orthofasciatus sp. nov., Minibiotus intermedius, Minibiotus poricinctus, Minibiotus floriparus and Minibiotus weglarskae.
Bertolanius portucalensis sp. nov.Figs. 3, 4
Material examined: Four specimens, one of them in simplex stage, and one egg from a moss sample collected in Amarante, northern Portugal (November 2005). Three specimens and the egg were mounted in Hoyer’s medium and one specimen in polyvinyl lactophenol.
Type repository: The holotype (slide N. 5234) and the egg (slide N. 5235) are deposited in the collection of Binda and Pilato (Museo del Dipartimento di Biologia Animale “Marcello La Greca”, Università di Catania); the three paratypes (slides BI-13 to BI-15) in the collection of P. Fontoura (Department of Zoology and Anthropology, Faculty of Sciences, University of Porto).
Specific diagnosis: Cuticle without pores but with very small round tubercles difficult to see; eye spots present; buccal tube wide; buccal armature with 14 peribuccal lamellae, an anterior band of tiny teeth difficult to see, a broad posterior band of teeth followed by a system of dorsal and ventral transverse ridges; the medio-dorsal and the medio-ventral ridges are subdivided into two large square-shaped teeth. Pharyngeal bulb with apophyses and three macroplacoids. Slender claws with well developed accessory points on main branches.
M. orthofasciatus M. intermedius M. poricinctus M. floriparus M. weglarskae
Diameter processes excluded 48 40–45 49 62 40
Diameter processes included 54 45–52 60 70 50
Processes around circumference 24 about 30 18–20 20–22 24
Processes in hemisphere about 60 about 130 50 60–75 about 50
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All claws with lunules, very large on the fourth pair of legs. Eggs with high membranous, finely reticulated ridges forming a reticulate design with large meshes on the egg shell.
FIGURE 3. A–D, Bertolanius portucalensis sp. nov.: A, detail of the cuticular tubercles (arrow) of the holotype; B, bucco-pharyngeal apparatus of the holotype; C, first pair of legs of the holotype, the thickened cuticular bar near the base of claws is shown; D, claws of the hind legs of the holotype (scale bars = 10 µm).
Description of the holotype: Body length 630 µm, colorless; eye spots present; cuticle without pores but with small round tubercles difficult to see (Fig. 3A). Buccal cavity terminal, very wide (25.0 µm), surrounded by fourteen peribuccal lamellae followed by a band of hardly visible tiny teeth. A well evident posterior ring of irregularly distributed teeth is also present (Fig. 3B); the teeth of the anterior row are well aligned. Behind
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that band of teeth a system of three dorsal and three ventral transverse ridges is present (Figs. 3 B, C). The latero-dorsal and latero-ventral ridges are elongated; the median ridges are subdivided into two robust square teeth (Fig. 3B). Rigid buccal tube 58.1 µm long and 12.1 µm wide externally (pt = 20.8). Stylet supports inserted on the buccal tube wall at 79.5 % of its length (pt = 79.5). Pharyngeal bulb with apophyses and three rod-shaped macroplacoids (Fig. 3B). Microplacoid absent. The first and the second macroplacoid are very close to each other. The first and third macroplacoid show the characteristic shape of one inverted “one” as described by Dastych (1972). The second macroplacoid is the shortest and the third the longest. First macroplacoid 10.6 µm long (pt = 18.2), second 10.2 µm (pt = 17.6), third 12.4 µm long (pt = 21.3); entire macroplacoid row 37.2 µm long (pt = 64.0).
FIGURE 4. A–D, Bertolanius portucalensis sp. nov.: A, egg; B, detail of the reticular design with large meshes formed by membranous ridges on the egg shell; C, D, details of veils formed by reticulated membranous ridges (arrows) (scale bars = 10 μm).
The claws, of Bertolanius type, (Figs. 3C, D), are slender, with well developed accessory points on the main branches; external and internal claws of the third pair of legs 20.1 µm long (pt = 34.6) and 18.3 µm (pt = 31.5) respectively. Posterior and anterior claws of the hind legs 24.1 µm long (pt = 41.5) and 19.6 µm long (pt= 33.7) respectively. Small, smooth lunules are present on legs I to III, larger and gently curled on legs IV (Fig. 3D). A thickened cuticular bar is present near the lunules on the legs I–III (Fig. 3C).
The eggs, freely laid, are spherical, with some high finely reticulated membranous ridges forming veils (Figs. 4C, D) connected to one another and therefore forming an evident reticular design with large meshes on the egg shell (Figs. 4A, B). At the margin of the egg, folds of the membranous ridges may seem to be conical processes, but a careful observation demonstrates that this is only an appearance. Diameter of the egg 93.5 µm without the membranous ridges, about 118 µm including these structures.
The paratypes are similar to the holotype in both qualitative and metric characters (Table 3). Etymology: The name portucalensis refers to the locus typicus, Portugal. Differential diagnosis: Only six species of the genus Bertolanius have been described until now. All of
them are extremely similar to each other, showing small differences in some characters of the buccal armature, claws and eggs. Some of them are very difficult to distinguish from one another without eggs, and some of them also have similar eggs.
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TABLE 3. Measurements (in µm) of some structures of the studied specimens of Bertolanius portucalensis sp. nov.
Bertolanius portucalensis sp. nov. differs from all the known species of the genus in the presence of very small cuticular round tubercles; the other species have smooth cuticle (only B. mahunkai (Iharos, 1971), according to the original description, has cuticular dots forming two longitudinal bands). B. weglarskae and, probably, B. mahunkai excluded, the new species also differs from the other species of the genus, in the characters of the egg.
In addition, the new species differs as follows from the other species of the genus: it differs from B. volubilis (Durante-Pasa and Maucci, 1975) in having three macroplacoids instead of two, width of the buccal tube almost uniform while in B. volubilis the anterior portion is clearly wider than the posterior one (see Durante Pasa & Maucci, 1975, Fig. 3), and both the medio-dorsal and medio-ventral ridges of the buccal armature subdivided into two large teeth.
It differs from B. nebulosus (Dastych, 1983) also in some characters of the buccal armature (both the medio-dorsal and medio-ventral ridges of the buccal armature are subdivided into two large teeth; the anterior teeth of the posterior band of teeth are larger, surely not smaller, than the posterior ones).
Bertolanius portucalensis sp. nov. differs from B. smreczynskii also in having shorter macroplacoids, the second macroplacoid shorter than the first one, and lunules on the legs IV curled.
It differs from B. markevichi (Biserov, 1992) also in having eye spots, buccal tube longer with respect to the body length and lower values of the pt index relative to the placoids and claws (Table 4), and lunules on the legs IV curled.
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The new species differs from B. mahunkai in having slightly wider buccal tube and the second macroplacoid shorter than the first one.
The new species differs from B. weglarskae in having wider buccal tube and both the medio-dorsal and medio-ventral ridges subdivided into two large teeth. The egg of Bertolanius portucalensis sp. nov. is very similar to that of B. weglarskae. Dastych (1972), in the description of this species, wrote that the veil is granulated. But in his monograph of 1988 (page 105) corrected this detail and specified: “veil covered with a network of very tiny meshes”. We examined eggs kindly send by this author collected from Tatry, and we confirm that the veil is finely reticulated.
Material examined: Three specimens from Amarante; one specimen from Parque Biológico (Avintes, Vila Nova de Gaia). All specimens were collected in moss samples and mounted in Hoyer’s medium.
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Body length 265.0 - 325.7 µm. The species could be easily confused with H. dujardini (Doyère, 1840), but the metric characters of the bucco-pharyngeal apparatus and claws obtained in the examined specimens match with those attributed to H. seychellensis.
Distribution: Up till now the species was recorded only in Seychelles Islands. This is the first record of this species in Europe.
Diphascon (Diphascon) pingue (Marcus, 1936)
Material examined: Eight specimens collected from moss samples at Amarante (one specimen) and at Parque Biológico (Avintes, Vila Nova de Gaia) (seven specimens). All the specimens mounted in Hoyer’s medium.
Body length 133.7–203.4 µm.Distribution: Very common species widely distributed in the whole world (McInnes, 1994), it may be
Material examined: Two specimens (body length 127.2 and 204.5 µm) collected from a moss sample at Parque Biológico (Avintes, Vila Nova de Gaia) were mounted in Hoyer’s medium.
Distribution: The species was recorded from various localities of the Mediterranean region (Italy, Turkey) and from North America.
TABLE 5. List of the species of limno-terrestrial tardigrades recorded from Portugal (Archipelagos of Madeira and Azores excluded). Year refers to the first record in the corresponding District. * = Described as a new species.
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Echiniscus multispinosus Da Cunha,
1944b*
Da CunhaFontoura
ViseuPorto
1944b
1981
Echiniscus oihonnae Richters, 1903 Da Cunha FontouraMaucci & Durante-Pasa
Coimbra, ViseuBragaGuarda
1944a
1982
1984a
Echiniscus quadrispinosus (Richters, 1902)
Da CunhaFontouraMaucci & Durante-PasaFontoura et al.
CoimbraBragaBejaPorto
19411982
1984c
this paper
Echiniscus scabrospinosus Fontoura 1982*
FontouraPilato et al.
CoimbraPorto
19822008
Echiniscus spinulosus (Doyère, 1840) Maucci & Durante Pasa Coimbra 1984c
Echiniscus testudo (Doyère, 1840) Fontoura Porto 1981
Echiniscus trisetosus Cuénot, 1932 Da CunhaMaucci & Durante-PasaMaucci & Durante-Pasa
Viana do Castelo, Castelo BrancoPortalegre, GuardaBragança, Évora
1947a
1984a
1984c
Hypechiniscus gladiator (Murray, 1905) Da Cunha Viana do Castelo 1947b
Pseudechiniscus suillus (Ehrenberg, 1853) Da Cunha
Fontoura Maucci & Durante-Pasa
Porto, Guarda, Coimbra, Castelo BrancoViseuBragaViana do Castelo, Vila Real, Portalegre, Évora
1941
1944a
1981
1984c
Pseudechiniscus facettalis Petersen, 1951 Maucci & Durante-Pasa Vila Real 1984c
EUTARDIGRADA
Murrayon hibernicus (Murray, 1911) Da Cunha Coimbra 1948
Murrayon pullari (Murray, 1907a) Da Cunha Viana do Castelo 1947a
Dactylobiotus macronyx (Dujardin, 1851) species inquirenda
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Macrobiotus orcadensis Murray, 1907a Fontoura Porto 1981
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Conclusion
After this study, 63 limno-terrestrial tardigrade species are recorded for Portugal, including 10 described as new species to science (Table 5). However, some records needs to be confirmed. Eventual mis-identifications,
Isohypsibius prosostomus Thulin, 1928 Da CunhaMaucci & Durante-Pasa
Fontoura et al.
Castelo BrancoGuardaVila RealPorto
1948
1984a
1984c
this paper
Isohypsibius sattleri (Richters, 1902) Da Cunha Coimbra 1947a
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already stressed by Maucci & Durante-Pasa (1984c), could have occurred. Macrobiotus echinogenitus
Richters, 1904 was probably confused with M. harmsworthi Murray, 1907a. Da Cunha (1941) described the observed specimens attributed to that species as having three macroplacoids instead of two. Some years later
and without other comments, Da Cunha (1947b) reported the presence of the species in other Portuguese stations. The first records of Macrobiotus harmsworthi and M. richtersi Murray 1911 are also doubtful because Da Cunha (1948) reported the synonym name M. schultzei Greeff, 1866. Fortunately, both species have been recorded in Portugal some years later (Fontoura, 1981). Echiniscus bisetosus Heinis, 1908, according to Maucci (1985), is not a valid species and the specimens attributable to this species belong to other species of the blumi group (E. blumi Richters, 1903; E. canadensis Murray, 1910; E. mediantus Marcus, 1930; E. trisetosus Cuénot, 1932), all of them already recorded in Portugal. According to Binda & Guglielmino (1982) Echiniscus bisetosus, and other elements of the series blumi-canadensis have probably to be attributed to E. blumi. Very probably, Bryodelphax tatrensis (Węglarska, 1959) is present in Portugal (the description and pictures of Bryodelphax parvulus Thulin, 1928 presented by Da Cunha (1941) seems to match with B. tatrensis). If so, the first Portuguese record of B. parvulus must be attributed to Maucci & Durante-
Pasa (1984c). On the other hand, Table 5 also shows that some Districts are very poorly surveyed and the number of Portuguese species will be probably much higher.
Acknowledgments
We want to thank Nuno Gomes Oliveira, the Director of the Parque Biológico de Gaia, for sampling facilities. We also thank the Museo Nacional de Ciencias Naturales (Madrid) and the colleagues Miguel Vilhena and Javier Sanchez who kindly sent type material of Minibiotus gumersindoi on loan, and the Museum of New Zealand (Wellington) for lending us specimens of Minibiotus bisoctus. We also thank H. Dastych (University of Hamburg) who kindly sent specimens and eggs of Bertolanius weglarskae, and C. Beasley who kindly improved our English. This research was supported by the Fundação para a Ciência e Tecnologia (Eco-Ethology Research Unit - FCT-331/94) and also by the University of Catania (Fondo Ricerca d’Ateneo).
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