New species and records of tardigrades from a biological repository collection from the Sierra Nevada de Santa Marta, Colombia.

The Sierra Nevada de Santa Marta (SNSM) is a mountain range in northern Colombia, recognized for its high biological richness. To date, the existence of 27 species of tardigrades is known for this particular region. Morphological and morphometric analysis of 51 specimens, found in 16 samples of bryophytes and lichens collected at the SNSM, and deposited in the tardigrade collection of the "Centro de Colecciones Científicas de la Universidad del Magdalena" between 2011 and 2016 was carried out. The species Mixibius gibbosus sp. nov. is described based on the following main traits: presence of gibbosities (up to date never reported for any species of the genus Mixibius), isodiametric tubercles and relevant morphometric differences with respect to the most similar congeneric species. Additionally, the species Diphascon pingue pingue sensu lato, Hypsibius cf. allisoni and Adropion onorei are, for the first time, recorded for Colombia and the latter is recorded for the first time out of Ecuador. All records are new for the SNSM.

An exceptional event during the moulting process of an eutardigrade: two bucco-pharyngeal apparatuses present.

A specimen of Diphascon (Diphascon) procerum with two bucco-pharyngeal apparatuses (one of which should have been expelled during moulting process) is described, and that anomalous condition is discussed.

Mixibius parvus sp. nov. and Diphascon (Diphascon) ziliense sp. nov., two new species of Eutardigrada from Sicily.

Two new species, Mixibius parvus sp. nov. and Diphascon (Diphascon) ziliense sp. nov. are described from Sicily. Mixibius parvus sp. nov. has three macroplacoids and a microplacoid and differs from M. tibetanus, the only other known species of the genus with those characteristics, in having a wrinkled cuticle without true small tubercles, a shorter microplacoid, smaller claw pt index values, and in lacking a cuticular bar on the first three pairs of the legs.Diphascon (Diphascon) ziliense sp. nov. lacks eye spots, has a pharyngeal bulb with two macroplacoids and a septulum, and possesses lunules and cuticular bars on the legs. It is similar to D. (D.) ramazzottii and D. (D.) procerum but differs from them in characters of the cuticular ornamentation and, in addition, from D. (D.) ramazzottii in having lunules and slightly longer claws in proportion to the body size and to the buccal tube. The new species differs from D. (D.) procerum in having stouter claws with a wider common portion and with the main branches shorter in proportion to the total length of the respective claws.

Two new tardigrade species from Sicily.

Two new species of tardigrades are described from Sicilian moss samples: Macrobiotus insuetus sp. nov. and Diphascon (Diphascon) procerum sp. nov.        Macrobiotus insuetus sp. nov. is a species of the harmsworthi-group characterized by both posterior and anterior claws of the hind legs, which are different in shape from those of the first three leg pairs. The IV claws have extended basal tract where the branches are joined and the secondary branch breaks at near right angle to the primary branch and is clearly shorter than the main branch and the secondary branch of claws I-III. The eggs are not areolated and have conical processes with a reticular ornamentation.        Diphascon (D.) procerum sp. nov. has a delicate cuticular ornamentation of very small tubercles, almost dots; two macroplacoids and septulum are present; thin accessory points are present on the main branches of the slender claws; lunules are absent but the base of the external claws of the hind legs are enlarged and slightly indented; a cuticular bar is present near the internal claw of the first three leg pairs and two cuticular bars are present on the hind legs between the base of the claws and near the base of the anterior claw. 

A new, simplified, drying protocol to prepare tardigrades for scanning electron microscopy.

A new protocol for preparation of tardigrades for scanning electron microscope (SEM) analysis is proposed. The more conventional protocols require various steps and a long time to obtain good drying of water bears, together with specific and uncommon instruments (i.e., critical point dryer) or highly volatile toxic compounds (i.e., hexametildisilazane). The new protocol can be performed using few and simple instruments and materials, all easily accessible, and produces a high yield in terms of dried animals in excellent condition for the observation of external morphological structures with SEM. The acquired data exhibit considerable promise, and the proposed methodology shows potential for application to other meiofaunal groups, including small arthropods, nematodes, and rotifers. RESEARCH HIGHLIGHTS: Cheap, safe, and fast new method for Tardigrada preparation for SEM. With the new protocol, the number of animals required for SEM studies is minimized. New protocol is potentially applicable to the study of other meiofaunal soft-bodied taxa.

A new tardigrade Doryphoribius maasaimarensis sp. nov. (Eutardigrada: Hypsibiidae) from Kenya.

A new tardigrade, Doryphoribius maasaimarensis sp. nov., is described from a moss sample collected in Masai Mara Game Reserve, Kenya. The new species is characterized by having a reticulate dorsal cuticle with small tubercles; nine transverse rows of dorsal gibbosities (configuration IX:4-6-4-6-4-6-4-4-2); pharyngeal bulb with two macroplacoids and no microplacoid; claws with short and thin accessory points; small, smooth lunules under the claws. The new species is most similar to Doryphoribius zyxiglobus (Horning, Schuster & Grigarick, 1978). Both exhibit two macroplacoids, similar cuticular pattern and the same configuration of gibbosities. However, in Doryphoribius maasaimarensis sp. nov. the cuticular tubercles are less close, buccal tube slightly longer with respect to the body length, more gradual curvature of the buccal tube, different claws shape and thinner accessory points.

Ramazzottius thulini (Pilato, 1970) bona species and description of Ramazzottius libycus sp. nov. (Eutardigrada, Ramazzottidae).

Comparing the type series of Ramazzottius thulini with specimens of Ramazzottius oberhaeuseri, from the locus typicus, we confirmed there are morphological and metric differences between these two species. This evidence convinced us that R. thulini should be considered a bona species. A new species, Ramazzottius libycus sp. nov., is also described. Ramazzottius libycus sp. nov. differs from R. oberhaeuseri in having more slender internal claws I-III and anterior claws IV, and other metric differences relative to all claws. The new species differs from R. thulini in details of the cuticular ornamentation, stylet supports inserted on the buccal tube in a slightly more caudal position, internal claws slightly different in shape, and some metric differences relative to all claws. It appears R. thulini and R. libycus sp. nov. are sibling species of R. oberhaeuseri and therefore the authors expect that there are other cryptic species included under the R. oberhaeuseri name still awaiting to be recognized and described.

Description of a new genus and species (Eutardigrada: Richtersiidae) from Colombia, with comments on the family Richtersiidae.

The new genus of the macrobiotoid family Richtersiidae Guidetti, Rebecchi, Bertolani, Jönsson, Kristensen Cesari, 2016 Crenubiotus gen. nov. is described, based on Crenubiotus crenulatus comb. nov. (Richters, 1904a) and on Colombian specimens formerly attributed to Macrobiotus cf. crenulatus and here described, after re-examination, as Crenubiotus revelator sp. nov. The new genus is characterised by an additional ventral thickening on the anterior portion of the ventral lamina, dorsal apophysis absent or very reduced, modified stylet furcae, claws equal in structure to those of Richtersius Pilato Binda, 1989, and very probably also Adorybiotus Maucci Ramazzotti, 1981 and Diaforobiotus Guidetti, Rebecchi, Bertolani, Jönsson, Kristensen Cesari, 2016; and, in the known species, cuticular pores, two macroplacoids, large well dentate lunules and a characteristic egg. The new species Crenubiotus revelator sp. nov. differs from Crenubiotus crenulatus comb. nov. in having smaller cuticular pores, a medio-ventral tooth in the buccal armature, more slender buccal tube, slightly more caudal stylet supports, slightly more slender claws, and in details of the egg. The description of the new genus, and new observations on Adorybiotus, Richtersius and Diaforobiotus, allowed the authors to complete the morphological scenario within the family Richtersiidae giving further support to such family.

Two new tardigrade species (Echiniscidae and Doryphoribiidae) from the Salta province (Argentina).

Tardigrades from Argentina are poorly known. Two new species from the Salta province are described in the present contribution: Pseudechiniscus saltensis sp. nov., and Doryphoribius cephalogibbosus sp. nov. The former species belongs to the novaezeelandiae group and, apart from the nominal species of the group, it is similar to nine congeneric species by the following characters: the absence of spines on legs I and lateral papillae, the scapular plate with a transversal fold, the presence of projections at the posterior margin of the pseudosegmental plate, caudal plate not faceted, and cuticular ornamentation in the form of heads (capituli) of endocuticular pillars, protruding through epicuticle and joined by striae. However, the new species differs from all of them in important qualitative (e.g. colour, different division of plates, details of cuticular ornamentation, claws etc.) and morphometric characters. The new species exhibits a unique morphology of gonopore and anus. Doryphoribius cephalogibbosus sp. nov., by the presence of two macroplacoids, dorsal gibbosities, and inhabiting terrestrial environment, belongs to the flavus group and differs from all members of the group in having ten, instead of nine, rows of gibbosities (X:2-4-6- 6-6-6-6-4-2-2+2[LI-III]), with as a peculiar exclusive character, the presence of the two cephalic gibbosities, never reported in other Doryphoribius species. Moreover, the new species differs from the other species of the group by morphometric and more detailed morphological characters. The new species description also gave an occasion to discuss the issue of gibbosity arrangement in other species and address the morphology of claw accessory points.

A new species of Isohypsibiid (Eutardigrada) from Sicily.

Dastych (2016) analysed the holotype of Isohypsibius tetradactyloides (Richters, 1907) and noticed that it possesses a very faint cuticular sculpture. For this and other characters, Dastych considered the species to be a synonym of Isohypsibius asper (Murray, 1906). The authors re-examined specimens attributed to Isohypsibius tetradactyloides by Pilato (1971) from two Sicilian rivers, and noticed that they cannot be attributed to Isohypsibius asper but to a new species. This new species belongs to the I. asper group, and is described here as Isohypsibius occultus sp. nov.. As the very faint cuticular ornamentation can be difficult to observe, the authors compared the new species with species having smooth cuticle or ornamented cuticle. Species with cuticular gibbosities or annulations, or clearly different in other characters, i.e. those with only two macroplacoids, the presence of a septulum, and cuticular bars on the legs, were excluded from comparison.

Meplitumenaluna gen. nov., sp. nov. an interesting eutardigrade (Hypsibiidae, Itaquasconinae) from the Sierra Nevada de Santa Marta, Colombia.

A new genus of Itaquasconinae, Meplitumen gen. nov., and a new species, Meplitumenaluna sp. nov., are described. The new genus has characters present in other genera of Itaquasconinae but in a unique combination. The spiral thickening of the bucco-pharyngeal tube is also present anteriorly to the insertion point of the stylet supports, excluding only the short portion where the apophyses for the insertion of the stylet muscles (AISM) are present. This character is similar to Astatumen Pilato, 1997 but Meplitumen gen. nov. differs from this genus as stylet furcae are shaped differently and as stylet supports and placoids are present. The presence of a spiral thickening in a portion of the buccal tube anterior to the stylet supports distinguishes the new genus from Mesocrista Pilato, 1987, Platicrista Pilato, 1987 and Itaquascon de Barros, 1939. Meplitumen gen. nov. also differs from Mesocrista, in having the caudal processes of the AISM pointing laterally (instead of postero-laterally), and the apices of the caudal processes of the stylet furcae unswollen. From Itaquascon the new genus also differs by having more robust stylet supports, pharyngeal bulb with placoids, stylet furcae differently shaped. Meplitumen gen. nov. also differs from Platicrista in having caudal processes of the AISM more robust and not flexible, and more slender stylet supports. The new species, Meplitumenaluna sp. nov., has a cuticle with a very faint roughness at the caudal extremity of the body, and eyes probably absent. The pharyngeal bulb is long, with two long, narrow, rod-shaped macroplacoids; a microplacoid and septulum are absent. The claws are well developed with main branches provided with accessory points, and at the base of the claws, a structure interpretable as a very thin lunule is present. Other cuticular thickenings on the legs are absent.

Echiniscidae from the Sierra Nevada de Santa Marta, Colombia, new records and a new species of Bryodelphax Thulin, 1928 (Tardigrada).

Three species of Echiniscus are recorded for the first time from Colombia: Echiniscus dariae, Echiniscus kofordi, and Echiniscus perarmatus. In addition, the description of the new species Bryodelphax kristensenisp. n., is mainly based on the presence of ten paired plus two unpaired granularly sculptured ventral plates, the dorsal plate ornamentation with superficial irregular pores, no spine on the anterior legs, and the hind legs without papillae or dentate collar.

Macrobiotus nebrodensis and Adropion vexatum, two new species of Eutardigrada (Tardigrada, Parachela) from Sicily.

Two new species of Eutardigrada are described from Sicily: Macrobiotus nebrodensis sp. nov. and Adropion vexatum sp. nov. The former species, belongs to the Macrobiotus hufelandi group, and is characterised by having a small, faint, microplacoid, and variable egg processes (most processes are shaped in the form of an inverted goblet with jagged terminal disc, but others are long, or very long, with a very small terminal disc). Adropion vexatum sp. nov. is characterised by having smooth cuticle, steep, vertical front to the head, pharyngeal tube longer than the pharyngeal bulb, which has small apophyses and three rod-shaped macroplacoids (microplacoid and septulum absent); long placoid row, about half the length of the pharyngeal bulb; main branches of claws with accessory points; lunules and other cuticular thickenings absent on the legs (small dots excluded).

New records of tardigrades from Colombia with the description of Paramacrobiotus sagani sp. nov. and Doryphoribius rosanae sp. nov.

By examining material collected in the Sierra Nevada de Santa Marta (Department of Magdalena, Colombia), the authors report a new record for the entire South America, Doryphoribius gibber Beasley & Pilato, 1987, and two species new to science, Paramacrobiotus sagani sp. nov., and Doryphoribius rosane sp. nov., are described. Paramacrobiotus sagani sp. nov. belongs to the richtersi group, vanescens subgroup (by having sculptured egg areolae) and is characterized by a peculiar cuticular granulation, trunco-conical egg processes with cylindrical indented apices, and other more detailed cha-racters both qualitative and metric; by the unique combination of characters, it differs from all the other known species of the genus. Doryphoribius rosanae sp. nov. is characterized by reticulated dorsal cuticle with gibbosities (formula IX:4-6-2-6-2-6-4-2-2), two macroplacoids in the pharynx without microplacoid or septulum, and large, stout claws without "free" accessory points but with lunules. It differs from all the other Doryphoribius species with gibbosities by having a unique formula, as well as other more detailed characters. Thanks to this contribution, the number of tardigrade species known for Colombia increases from 52 to 55.

Milnesium minutum and Milnesium sandrae, two new species of Milnesiidae (Tardigrada, Eutardigrada, Apochela).

Two new species of Milnesium are described, Milnesium minutum sp. n. from Sicily and Milnesium sandrae sp. n. from the Hawaiian Archipelago. The body size of Milnesium minutum is the smallest of the known species of the genus. The stylet supports are inserted on the buccal tube at 63-66% of its length and the claws have a [3-3]-[3-3] configuration. Milnesium sandrae has stylet supports inserted on the buccal tube at 58-60.5% of its length, a [3-3]-[3-3] claw configuration, and the percent ratio between the secondary claw and primary claw length on legs I-III (78.6%-85.5%) clearly higher than on legs IV (70.5%-71.4%). With the description of these two new species, the number of species in the genus is increased to 31.

Two new species of Milnesium (Tardigrada: Milnesiidae).

Two new species of Milnesium are described: Milnesium vorax sp. nov from Sicily and Milnesium tumanovi sp. nov. from Crimea. Milnesium vorax sp. nov. is uncoloured, with eye spots, smooth cuticle, wide buccal tube, stylet supports inserted on the buccal tube at almost 62 % of its length; claws with configuration [2-3]-[3-2]; accessory points present on the main branches. Milnesium tumanovi sp. nov. is also uncoloured, has eye spots, smooth cuticle, and wide buccal tube, but it has the stylet supports inserted on the buccal tube at about 52 % of its length, and claws with configuration [3-3]-[3-3]; the accessory points are present on the main branches.

Eremobiotus ginevrae sp. nov. and Paramacrobiotus pius sp. nov., two new species of Eutardigrada.

Two new eutardigrade species are described: Eremobiotus ginevrae sp. nov. and Paramacrobiotus pius sp. nov. The first is similar to Eremobiotus alicatai (Binda, 1969) but differs in the claw shape and dimensions. It has been found in Sicily, Israel and Russia. The second species, belonging to the richtersi group, is currently found exclusively in Sicily. It has a smooth cuticle, three macroplacoids and a microplacoid, eggs with reticulated trunco-conical processes with small terminal thorns; the egg-shell is areolated and the areolae are clearly sculptured.

Two new freshwater eutardigrade species from Sicily.

Two new species of freshwater Eutardigrada are described from Sicily: Isohypsibius rusticus sp. nov. and Isohypsibius zappalai sp. nov. The former species has eye spots, ornamented body surface with many, variously sized tubercles; bucco-pharyngeal apparatus of the Isohypsibius type; pharyngeal bulb with apophyses and two rod-shaped macroplacoids; microplacoid absent; claws, of the Isohypsibius type, well developed, with long and thin common basal portion; main branches of all claws without free accessory points; very thin lunules present; cuticular bars on the legs absent. Isohypsibius zappalai sp. nov. has eye spots; entire body surface with small tubercles rounded in shape, fairly uniformly sized and tending to form transverse lines; bucco-pharyngeal apparatus of the Isohypsibius type, pharyngeal bulb with apophyses and two macroplacoids; microplacoid absent; claws of the Isohypsibius type, well developed, with long common basal portion and both main and secondary branches with a wide proximal portion. Main branches of all claws with accessory points; small, flexible lunules present; cuticular bars on the legs absent.

Tardigrada from a sub-Andean forest in the Sierra Nevada de Santa Marta (Colombia) with the description of Itaquascon pilatoi sp. nov.

Currently only 32 species of limno-terrestrial tardigrades have been reported in the literature for Colombia. Our study focused on both heterotardigrades and eutardigrades, which were extracted from eight samples of bryophytes and lichens collected in a sub-Andean forest transect in the Sierra Nevada de Santa Marta, Colombia. Fourteen species were found, six of which are new records for Colombia: Echiniscus madonnae Michalczyk & Kaczmarek, 2006, Echiniscus virginicus Riggin, 1962, Milnesium krzysztofi Kaczmarek & Michalczyk, 2007, Doryphoribius amazzonicus Lisi, 2011, Isohypsibius sattleri (Richters, 1902) and Diphascon higginsi Binda, 1971; and one new to science. Itaquascon pilatoi sp. nov., is characterized by having smooth cuticle, no eyes, buccal tube almost as long as the pharyngeal tube, well developed, obvious stylet furcae with long branches, slender claws, no lunules and no cuticular bars on the legs. The new species differs from I. umbellinae Barros, 1939, the most similar species, in having the stylet supports inserted precisely at the border between buccal and pharyngeal tube, more slender claws and more pronounced length differential between the external and internal claws of each leg. The total number of Colombian limno-terrestrial tardigrade species is raised to 37. 

Persistence of phlebotomine Leishmania vectors in urban sites of Catania (Sicily, Italy).

BACKGROUND: Pioneering research on "Mediterranean Kala-Azar" carried out by Adler and Theodor early in the past century (~1930s) had identified Catania city (Sicily) as a major focus of the disease nowadays known as zoonotic visceral leishmaniasis (VL). Despite the fact that disease in both humans and dogs has continued to be highly prevalent in the Catania province up to the present times, research on Leishmania vectors in this urban focus dates back to that distant period. This study aimed to evaluate the persistence and current composition of the sand fly fauna in urban environments of Catania in recent years, 2006 and 2013. METHODS: In 2006 fifty-one suitable collecting sites were identified within 44 sub-units of a grid drawn to include the urban Catania area. In 2013 the survey was restricted to four of the most productive and representative sites resulting from the 2006 survey. In both periods 3 collections per month were performed using standard sticky traps set for 3 days in wall holes/cavities along public roads, from the end of April through December. RESULTS: 43/51 sites (84.3%) were found positive for sand flies. The 2006 collections accounted for a total of 4341 specimens including six species. Among competent Leishmania vector species, P. perniciosus was the most prevalent (36.5%) being identified in all sand fly-positive sites, with significant abundance in those of the old city centre. Other species of interest were P. sergenti (2.5%) and P. neglectus (1.5%). The 2013 survey produced 1130 sand flies, of which 39.5% were P. perniciosus, 1.6% P. sergenti and 0.7% P. neglectus. A search for Leishmania DNA in a small sample of 72 P. perniciosus females revealed 11% infection prevalence. CONCLUSIONS: Our findings from an old urban focus of leishmaniasis demonstrate that phlebotomine sand flies have adapted fairly well to the drastic environmental changes that have occurred in cities of the Western world in the past century and still represent a potential risk for Leishmania transmission.