Biomass variation and reproductive phenology of Gracilaria gracilis in a Patagonian natural bed (Chubut, Argentina)

The biomass variation and the reproduction of the natural Gracilaria gracilis bed in Bahía Bustamante (Patagonia, Argentina) were analyzed for 2 years, with the aim of determining the present situation of the population for an updated status overview; establishing the relevant features of the temporal variation in both biomass and reproductive states in relation to environmental factors, epiphytes and associated algae; and assessing carpospore availability for future spore-culture development. Field measurements and sampling were performed monthly between March 2006 and February 2008. In both years, G. gracilis biomass presented marked seasonal variations, with a minimum in winter and a maximum in late spring and in summer. During both years, coexistence of the three life-cycle phases was found, with dominance of tetrasporophytes. Two data sets from individuals originated from sexual reproduction (tetraspores and carpospores) and from asexual reproduction by thallus fragmentation were analyzed separately. In the fragmentation fraction, tetrasporophyte frequencies remained higher than those for gametophytes. However, in the spore-originated fraction, a generation ratio close to 0.5 was observed. Female gametophytes bearing cystocarps were always present, with a maximum in summer and autumn. Biological data were related to environmental factors by means of canonical correspondence analysis (CCA). The first year was characterized by higher biomass values of G. gracilis and Undaria pinnatifida, lower epiphytism, larger Gracilaria thalli and greater proportion of mature tetrasporophytes and gametophytes. The second year was characterized by a high proportion of Gracilaria vegetative thalli and high epiphyte density. The best time to obtain spores from cystocarpic thalli would be in summer and early autumn.     

Seasonal variation in agar composition and properties from Gracilaria gracilis (Gracilariales,Rhodophyta) of the Patagonian coast of Argentina

Seasonal variation of agar from specimens of a commercially exploited population of Gracilaria gracilis (Stackhouse) Steentoft, Irvine & Farnham in the Patagonian coast of Argentina was studied. For each seasonal harvest, random samples of plants were pooled for subsequent polysaccharide extraction at different water temperatures and agar physico‐chemical properties and composition were determined. Both spring and summer plants yielded 30% and 41% of agar, respectively, which differed slightly in their at rest rheological behavior. Spring and summer plants produced strong gels (238 and 218 g cm^?2, respectively), but the latter gels had nil adhesiveness. In autumn plants, agar yield decayed to 19%, though the product still maintained similar gel strength (210 g cm^?2). Adhesiveness in this product was at least an order of magnitude higher than in the others, concomitant with a peak in the formation of tetraspores and carpospores. This suggests a biological role for the galactan in the initial attachment of spores to the substrate. But since fall corresponds to the settling of reproductive structures, caution should be taken to harvest the algae once spores have been shed from mother plants.     

Succession in subtidal macrofouling assemblages of a Patagonian harbour (Argentina, SW Atlantic)

Subtidal fouling assemblages usually consist of short-lived organisms. Colonisation sequences on man-made structures may thus be greatly affected by the temporal and spatial variability of propagule supply. This study explores the influence of seasonality on succession in the macrofouling assemblage of a Patagonian harbour (Argentina, Southwest Atlantic). Replicated artificial substrata were suspended horizontally and sampled at quarterly intervals during 1?year. The influence of seasonality on 1-year-old assemblages was further analysed using additional sets of replicated panels submersed at different seasons and collected 1?year later. Upper surfaces were always dominated by ephemeral algae, while lower surfaces exhibited high coverage of filter-feeding invertebrates. Regardless of submersion length, species richness was significantly higher on lower than on upper surfaces. A significant interaction between orientation and submersion length was found for the Shannon diversity index, meaning that temporal changes in diversity depended on substratum orientation. On the lower surfaces, diversity reached a maximum after 9?months and then declined, mainly due to extensive dislodgment of two species of ascidians. On algal-dominated upper surfaces, differences in structure of annual assemblages were due to seasonal changes in the abundance of ephemeral algae. This study shows that constancy or variability of 1-year-old assemblages whose development began at different seasons depended greatly on the life history of the organisms that settled and managed to persist on both surfaces, which in turn depended on substratum orientation.     

Agar from Gracilaria gracilis (Gracilariales, Rhodophyta) of the Patagonic coast of Argentina--content, structure and physical properties

Milled summer thalli of Gracilaria gracilis from Argentina were sequentially extracted with water at room temperature (RTW1-3), 70 degrees C (W701-3) and 90 degrees C (W901-2). Both W701 and W901 consisted of high molecular weight polysaccharides (ca. 540,000Da), but polydispersity was higher for the major product W701 (yield, 72% of the recovered). Structural analyzes by methylation and (13)C NMR spectroscopy revealed that W701 was mainly agarose. Alkaline treatment, together with structural analyzes, indicated a negligible proportion of precursor l-galactose 6-sulfate residues in this product, while they were clearly detected in the (13)C NMR spectra of RTW2-3. The presence of floridean starch in W901 had an antagonistic effect on its gel strength, which resulted nearly three times lower than that of fraction W701. Ultrastructural observation by transmission electron microscopy showed that, after extraction with hot water, a partial loss of cell wall stratification and disorganization of the cuticle had occurred. Final cellular debris exhibited swelling in the microfibrillar component. After this first thorough study of the chemical composition and physical properties of the products of G. gracilis from Bahía Bustamante we conclude that a good quality agarose is obtained in high yield after extraction with water at 70 degrees C without the requirement of alkaline pretreatment, which usually produces degradation of the polysaccharide.     

Massive corals in Paleocene siliciclastic sediments of Chubut (Argentina)

A horizon with large, massive corals in growth position was discovered in the Paleocene, probably upper Danian, part of the Maastrichtian–Paleocene Lefipán Formation of Chubut (Patagonia, Argentina). All corals belong to one species, the cosmopolitan Haimesiastraea conferta Vaughan, which survived the end-Cretaceous mass extinction. The occurrence of massive corals at this site is exceptional both because of the siliciclastic depositional regime and because of the high palaeolatitude setting. An unusual autecology of this coral and strongly reduced sedimentation rates, were probably the prerequisites for coral growth, but a link to palaeoclimate is less likely.     

Palynological characterization of honeys from the Andean Patagonia (Chubut,Argentina)

The pollen content of 58 honey samples of Apis mellifera L. from the Andean region of Chubut (Argentinian Patagonia) was analyzed. The samples were provided by beekeepers between 1999 and 2004. Eighty‐eight pollen types were identified, 30 of them are not reported in Argentinian honeys. Identified types belong to 47 families, of which the most diverse were Asteraceae (15 pollen types), Fabaceae (13 pollen types), and Rosaceae (four pollen types). From the samples analyzed, 47% were monofloral and corresponded to the following taxa: Trifolium spp. (16%), Rosaceae (10%), Aristotelia chilensis (Molina) Stuntz (7%), Discaria‐Colletia (5%), Escallonia spp. (3%), Schinus patagonica (Phil.) I. M. Johnst. (2%), Phacelia secunda J. F. Gmel. (2%), Myrtaceae (2%). The results of this study show a high level of utilization of native flora by Apis mellifera. Four new types of monofloral honey were recognized: Aristotelia chilensis, Discaria‐Colletia, Phacelia secunda, and Schinus patagonica. The high representation of native flora distinguishes these honeys geographically.     

Seasonal and spatial distribution of stoneflies in the Chubut River (Patagonia, Argentina)

Longitudinal and seasonal distributions of Plecoptera species were examined along the Chubut River in the Patagonian Precordillera and Plateau, Argentina. Chubut River (>1000 km) is the largest river in the area and the hydrological regime is modified in the lower section by an artificial reservoir (Florentino Ameghino Dam). Quantitative samples were collected in 13 sites in the higher, middle, and lower sections of the river basin. Sites were visited four times during 2004, and a total of nine species and 5772 individuals were collected in the study. Plecoptera richness decreased dramatically from the headwaters to the mouth of the river system. Two species, Antarctoperla michaelseni and Potamoperla myrmidon, were able to live below the impoundment but they were not abundant. Notoperlopsis femina and A. michaelseni abundances were higher in summer than in the other seasons, while Limnoperla jaffueli peaked in spring. Species–environmental relationships were examined using Canonical Correspondence Analysis and six independent variables were identified as the major factors structuring stoneflies assemblages. First axis was highly related to environmental variables reflecting the hydro-geological and land use gradients in the basin (conductivity, total suspended solids, periphyton Chlorophyll a). Second axis was more related to variables that changed seasonally (wet width, water temperature and soluble reactive phosphate).     

Non-random mating in controlled multiple-donor crosses in Gracilaria gracilis (Gracilariaceae,Rhodophyta)

Crosses were performed to identify the sources of variation in zygote production (via cystocarp production) in Gracilaria gracilis, a red haploid-diploid seaweed. First, because male gametes are short-lived (<6?h), the rate of gamete encounters was evaluated in a time-course experiment. Second, the effect of water motion on gamete encounters was assessed by introducing turbulent eddies in the crossing tank and by comparing fertilization rates with and without this added turbulence. Third, variation due to individual performance was explored by performing multiple-donor crosses using 12 males and 12 females from three populations. Paternity of cystocarps produced in these crosses was determined using microsatellite markers. The results show that cystocarp yield increased with exposure time: fertilization occurred in as little as 15?min after the introduction of male branches into the crossing tank and maximum cystocarp production values were observed at 6?h. There were no significant differences in cystocarp production between the two turbulence levels. On the other hand, cystocarp production was highly influenced by male and female parental identities and to a lesser degree by an interaction between the male and female parents. The variation in cystocarp production according to male and female identity was not due to population origin as there was no difference between intra- and inter-population crosses. Thus nonrandom mating occurs in controlled conditions and arises from differential performance in G. gracilis. There was a strong deviation from equality of male performance, implicating post-adhesion events and/or male gamete production as important in generating non-random mating. Consequently, non-random mating may play a role in the evolution of mating patterns in G. gracilis.     

New primate fossils from late Oligocene (Colhuehuapian) localities of Chubut Province, Argentina

New primate fossils have been recovered from the late Oligocene (Colhuehuapian) localities of Gaiman and Sacanana in Patagonian Argentina. The new fossils are provisionally allocated to Dolichocebus gaimanensis and Tremacebus harringtoni, the only primates previously described from these localities. These new dental remains are more primitive than the teeth of any previously known platyrrhines, living or fossil, and conform extremely well with the hypothetical ancestral morphotype for New World monkeys suggested by several authors. They are also very similar to the teeth of Oligocene catarrhines from Egypt such as Aegyptopithecus zeuxis.     

Colonization of subtidal macroalgae in a fucalean-dominated algal assemblage,southwestern Australia

The influence of fucalean canopy species and dominant understory macroalgae on algal colonization was investigated to evaluate whether layering contributes to patterns in algal diversity. Patterns in recruitment were compared among total-clearing, understory-removal, canopy-removal, and undisturbed plots (plot area = 0.25 m²), using a randomized block design in depths <10 m and 10–20 m at Woody Island, Western Australia. To evaluate if propagules were available in the water column above the canopy layer, settlement plates (plate area = 0.04 m²) were deployed in depths <10 m, 10–20 m, and >20 m. A total of 198 macroalgal species was recorded. Biomass of the understory species Osmundaria prolifera Lamouroux and Botryocladia sonderi Silva was similar between canopy-removal and undisturbed plots. Diversity of macroalgae was similar in the presence and absence of a canopy layer. Taxa found in the canopy showed different patterns in recruitment. Cystoseiraceae recruited predominantly in total-clearings in both depth strata. Sargassaceae recruited most abundantly in depths <10 m. Density of canopy taxa on settlement plates was similar with depth (20–30 juveniles per plate), and juveniles were mainly Cystoseiraceae. In contrast to kelp beds or forests, patterns in algal colonization appeared to be maintained by environmental factors or processes other than the direct effects of layering in the subtidal fucoid-dominated assemblages at Woody Island. Handling editor: K. Martens     

Ecological characteristics of Triatoma patagonica at the southern limit of its distribution (Chubut, Argentina)

This study aimed at identifying the ecological characteristics of Triatoma patagonica Del Ponte 1929 (Hemiptera: Reduviidae) and evaluating the epidemiological importance of this species at its southernmost distribution limit. We carried out two surveys in the Province of Chubut, in summer and in spring, 1998. In each survey, we interviewed local health care agents for triatomine recognition and reports of the bugs, followed by entomological searches in houses and peridomestic and silvatic biotopes. The presence of T. patagonica was confirmed in two of the five localities indicated by interview. In agreement with previous studies, all these positive localities were east of the 11 degrees C isotherm, within the Monte phytogeographic province. Triatomine abundance and infestation levels (in peridomiciles and peridomestic biotopes) were higher in summer than in spring, possibly reflecting adverse environmental conditions such as flood and cold winter weather. In the silvatic environment, we found three adult bugs under rocks in summer. In peridomestic sites bugs were only found associated with chickens, which were also the only blood meal source identified. Infection by Trypanosoma cruzi was not detected. We conclude that T. patagonica at its southernmost distribution limit does not represent a risk to public health due to its low abundance, lack of association with humans, and absence of T. cruzi infection.     

Protoplast isolation optimization and regeneration of cell wall in Gracilaria gracilis (Gracilariales, Rhodophyta)

This paper reports the first successful isolation and cell wall regeneration of Gracilaria gracilis (Stackhouse) Steentoft, Irvine et Farnham protoplasts. These results form an important foundation for the development of a successful tissue culture system for G. gracilis. Initially, an isolation protocol was optimized by investigation of the effects of the enzyme constituents and concentrations, the pre-treatment of thalli, the incubation period and temperature, and the pH of the enzymatic medium on protoplast yields. A pre-treatment of G. gracilis thalli with 1 % (w/v) papain for 30 min followed by a 3-h enzymatic digestion of thalli with an enzymatic mixture containing 2 % (w/v) cellulase Onozuka R-10, 1 % (w/v) macerozyme R-10, and 10 U mL^?1 agarase at pH 6.15 was found to produce the highest yield of protoplasts at 22 °C. Reliably high yields (20–30?×?10⁵ protoplasts g^?1 f.wt) of protoplasts could be obtained from G. gracilis thalli when this optimized protocol was used. Cell wall re-synthesis by G. gracilis protoplasts, which constitutes the first step towards whole plant regeneration, was followed using calcoflour staining and scanning electron microscopy. Protoplasts were shown to complete the initial stages of cell wall re-synthesis within the first 24 h of culturing.     

Late Miocene continental biota in Northeastern Patagonia (Península Valdés, Chubut, Argentina)

A new vertebrate faunal assemblage was recently discovered from the uppermost part of the late Miocene Puerto Madryn Formation. These deposits crop out along the southwestern coast of the Península Valdés area near Punta Delgada (Chubut Province, Argentina). The exhumed vertebrate fauna includes a range of fish, bird and mammal taxa, of which the latter are most varied and abundant. The new findings represent the first record of continental fossil vertebrates in the Puerto Madryn Formation and this is the first assemblage of late Miocene continental vertebrates recorded to the south of Río Negro Province. It also includes the southernmost record of Hydrochoeridae rodents, Dendrocygninae birds and Loricariidae fishes. The mammals suggest that the fossil-bearing sediments are Huayquerian in age. The climate during the accumulation of late Miocene deposits in this region is inferred to have been warmer and seasonally drier than that of today.     

Association between Hyperoche martinezii (Amphipoda: Hyperiidae) and ctenophores from the Buenos Aires coast,Argentina (South-western Atlantic Ocean)

This survey examined the association between the hyperiid amphipod Hyperoche martinezii and ctenophores off the Argentinian coast (38°08′17″S, 57°31′18″W) through the evaluation of seasonality, prevalence and intensity of infection during an annual cycle. Medusae were also examined but only the ctenophores Mnemiopsis leidyi, Pleurobrachia pileus and Beroe ovata showed this association during the austral mid-spring to mid-summer. A total of 502 hyperiids were obtained; most (422 individuals) were larval stages, 53 juveniles and 27 adults. Mnemiopsis leidyi had the highest number of hyperiids with 98.6%, followed by P. pileus (0.80%), and B. ovata (0.60%). Total prevalence was 2.0 and intensity of infection ranged between 1 and 17 hyperiids per ctenophore. The host with highest prevalence was B. ovata (4.54), followed by M. leidyi (3.76) and P. pileus (0.1). Prevalence values had some correlations with the increase in the total length of B. ovata (r?=?0.480, P?=?0.006) and M. leidyi (r?=?0.501, P?B. ovata (r?=?0.425, P?=?0.017). The hyperiid was found in different parts of the host body: larval stages were found in the canal close to the subtentacular comb row and the stomodeum, whereas juvenile/adult stages were observed with a resting posture on the external surface of the ctenophores. The known geographic distribution of H. martinezii was extended; this finding represents the addition of three new hosts for this hyperiid.     

Investigation of the Gracilaria gracilis (Gracilariales,Rhodophyta) proteome response to nitrogen limitation

Inorganic nitrogen has been identified as the major growth‐limiting nutritional factor affecting Gracilaria gracilis populations in South Africa. Although the physiological mechanisms implemented by G. gracilis for adaption to low nitrogen environments have been investigated, little is known about the molecular mechanisms of these adaptions. This study provides the first investigation of G. gracilis proteome changes in response to nitrogen limitation and subsequent recovery. A differential proteomics approach employing two‐dimensional gel electrophoresis and liquid chromatography–tandem mass spectrometry was used to investigate G. gracilis proteome changes in response to nitrogen limitation and recovery. The putative identity of 22 proteins that changed significantly (P < 0.05) in abundance in response to nitrogen limitation and recovery was determined. The identified proteins function in a range of biological processes including glycolysis, photosynthesis, ATP synthesis, galactose metabolism, protein‐refolding and biosynthesis, nitrogen metabolism and cytoskeleton remodeling. The identity of fructose 1,6 biphosphate (FBP) aldolase was confirmed by western blot analysis and the decreased abundance of FBP aldolase observed with two‐dimensional gel electrophoresis was validated by enzyme assays and western blots. The identification of key proteins and pathways involved in the G. gracilis nitrogen stress response provide a better understanding of G. gracilis proteome responses to varying degrees of nitrogen limitation and is the first step in the identification of biomarkers for monitoring the nitrogen status of cultivated G. gracilis populations.     

Intertidal and Subtidal Fouling Assemblages in a Patagonian Harbour (Argentina, Southwest Atlantic)

The composition of the early stages of intertidal and subtidal fouling assemblages in Comodoro Rivadavia harbour (Argentina, 45°52′ S, 67°28′ W) and the influence of shore level and season on their structure were analysed. At the beginning of each season, stones were glued to the substratum with epoxy putty and distributed along 4 vertical transects at intervals of 20 m, at 3 levels: upper intertidal, middle intertidal, and subtidal. Substrata remained in the field for 84–100 days. A total of 48 samples (4 seasons × 3 levels × 4 replicates) were analysed. Species richness increased with depth, with 6 taxa in the upper intertidal, 23 in the middle intertidal and 31 in the subtidal. Seasonal differences in richness were less distinct. Green, red and brown algae were the dominant groups. Invertebrates were mainly represented by filter-feeding, sessile organisms, such as cheilostome bryozoans, spirorbid polychaetes and acorn barnacles. The barnacle Balanus glandula and the bryozoan Cryptosula pallasiana were the only non-indigenous species found in this study. Highly significant differences in structure among shore levels and seasons were evidenced by a two-way ANOSIM test. The upper intertidal is characterized by the filamentous green algae Urospora penicilliformis and Ulothrix flacca. The barnacle Balanus glandula is the most abundant species in the middle intertidal. The subtidal is defined mainly by the presence of the spirorbid polychaetes Paralaeospira levinseni and Romanchella perrieri, and the keyhole limpet Fissurella radiosa. Ordination of samples by season was less clear than by shore level.