bluuebunny/crossref_metadata_2025 huggingface dataset:
Microheterogeneity in Ionic Liquid Mixtures: Hydrogen Bonding, Dispersed Ions, and Dispersed Ion Clusters
Authors: Mezzetta Andrea , Rodriguez Douton Maria J. , Guazzelli Lorenzo , Pomelli Christian Silvio , Chiappe Cinzia
Published: November 2018

Mixtures of ionic liquids (ILs) having a common ion but differing in the identity of the anion or cation represent highly interesting media. By varying the composition, one can successfully modulate specific physicochemical, structural, and biological properties. The molecular interactions (coulombic, hydrogen-bonding, van der Waals, and π–π intermolecular forces) that determine the three-dimensional structure of pure ILs can indeed be modified by the addition of another IL. In this context, we present here a 1H NMR, Fourier transform (FT)-IR, thermogravimetric, and solvatochromic study of the structural features of IL binary mixtures based on a common imidazolium cation ([CnC1im]+) and anions of different size and hydrogen-bond acceptor ability. For each mixture, the analyses were carried out at different molar ratios of the two components.
Evolution in the deep sea: a combined analysis of the earliest diverging living chitons (Mollusca : Polyplacophora : Lepidopleurida)
Authors: Sigwart Julia D. , Schwabe Enrico , Saito Hiroshi , Samadi Sarah , Giribet Gonzalo
Published: November 2018

Lepidopleurida is the earliest diverged group of living polyplacophoran molluscs. They are found predominantly in the deep sea, including sunken wood, cold seeps, other abyssal habitats, and a few species are found in shallow water. The group is morphologically identified by anatomical features of their gills, sensory aesthetes, and gametes. Their shell features closely resemble the oldest fossils that can be identified as modern polyplacophorans. We present the first molecular phylogenetic study of this group, and also the first combined phylogenetic analysis for any chiton, including three gene regions and 69 morphological characters. The results show that Lepidopleurida is unambiguously monophyletic, and the nine genera fall into five distinct clades, which partly support the current view of polyplacophoran taxonomy. The genus Hanleyella Sirenko, 1973 is included in the family Protochitonidae, and Ferreiraellidae constitutes another distinct clade. The large cosmopolitan genus Leptochiton Gray, 1847 is not monophyletic; Leptochiton and Leptochitonidae sensu stricto are restricted to North Atlantic and Mediterranean taxa. Leptochitonidae s. str. is sister to Protochitonidae. The results also suggest two separate clades independently inhabiting sunken wood substrates in the south-west Pacific. Antarctic and other chemosynthetic-dwelling species may be derived from wood-living species. Substantial taxonomic revision remains to be done to resolve lepidopleuran classification, but the phylogeny presented here is a dramatic step forward in clarifying the relationships within this interesting group.
Barcoding of mygalomorph spiders (Araneae : Mygalomorphae) in the Pilbara bioregion of Western Australia reveals a highly diverse biota
Authors: Castalanelli Mark A. , Teale Roy , Rix Michael G. , Kennington W. Jason , Harvey Mark S.
Published: November 2018

The Pilbara bioregion of Western Australia is an area that contains vast mineral deposits and unique ecosystems. To ensure that mineral deposits are mined with minimal impact on the natural environment, impact assessment surveys are required to determine what fauna and flora species are located within proposed development areas, in particular, by determining the distributions of short-range endemic species (SREs). One infraorder of Arachnida, the Mygalomorphae (trapdoor spiders and their kin), are frequently identified as SREs. These identifications are traditionally performed using morphological techniques; however, only males can be reliably identified to species. Furthermore, the majority of species have not been formally described and males comprise only ~5% of specimens collected. To assess mygalomorph diversity and the distribution of species in the Pilbara, we employed a molecular barcoding approach. Sequence data from the mitochondrial DNA cytochrome c oxidase subunit I (COI) gene were obtained from 1134 specimens, and analysed using Bayesian methods. Only a fraction of the total mygalomorph fauna of the Pilbara has been documented, and using a species boundary cut-off of 9.5% sequence divergence, we report an increase in species richness of 191%. Barcoding provides a rapid, objective method to help quantify mygalomorph species identifications and their distributions, and these data, in turn, provide crucial information that regulatory authorities can use to assess the environmental impacts of large-scale developments.
From Gondwana to Europe: inferring the origins of Mediterranean Macrothele spiders (Araneae : Hexathelidae) and the limits of the family Hexathelidae
Authors: Opatova Vera , Arnedo Miquel A.
Published: November 2018

The family Hexathelidae ranks among the smaller mygalomorph spider families. Most species are endemic to the Australasian region and the family was traditionally considered an example of a Gondwanan lineage. However, recent studies have cast some doubt on the monophyly of the family. Macrothele is the only genus with an out-of-Gondwana distribution. The bulk of the Macrothele diversity is found in South-east Asia, few species are known from central Africa and two species inhabit Europe: Macrothele calpeiana (Walckenaer, 1805) from the Iberian Peninsula and Macrothele cretica Kulczynski, 1903 endemic to Crete. Here we investigate the origins of the European Macrothele species by means of a multi-locus phylogenetic approach and by inferring the time frame of the diversification of the genus using Bayesian relaxed clock methods. We also provide further insights into the phylogenetic status of the family Hexathelidae. Our results indicate that the diversification of Macrothele traces back to the period of the Gondwana break-up and its present-day distribution most likely reflects the subsequent tectonic plate movements. The two European species were not recovered as sister taxa, suggesting that Macrothele colonised the Mediterranean region twice independently. The polyphyly of the family Hexathelidae is further confirmed and the subfamily Atracinae is identified as the conflicting lineage.
Tracing sewage-derived organic matter into a shallow groundwater food web using stable isotope and fluorescence signatures
Authors: Hartland Adam , Fenwick Graham D. , Bury Sarah J.
Published: November 2018

Little is known about the feeding modes of groundwater invertebrates (stygofauna). Incorporation of sewage-derived organic matter (OM) into a shallow groundwater food web was studied using fluorescence and stable isotope signatures (δ13C and δ15N). Organic pollution was hypothesised to limit sensitive species’ abundances along the contamination gradient and isotope signatures of stygofauna consuming sewage-derived OM were expected to be enriched in δ15N. Stygofauna communities near a sewage treatment plant in New Zealand were sampled over 4 months and microbial biofilms were incubated in situ on native gravel for 1 month. As anticipated, OM stress-subsidy gradients altered stygofauna composition: the biomass of oligochaetes and Paraleptamphopus amphipods increased in OM-enriched groundwater (higher dissolved organic carbon (DOC) and tryptophan-like fluorescence), whereas other, probably less-tolerant taxa (e.g. ostracods, Dytiscidae) were absent. Isotopic signatures for stygofauna from polluted groundwater were consistent with assimilation of isotopically enriched sewage-N (δ15N values of 7–16‰), but highly depleted in δ13C relative to sewage. Negative 13C discriminations probably occur in Paraleptamphopus amphipods, and may also occur in oligochaetes and Dytiscidae, a finding with implications for the application of δ13C for determining food sources in groundwaters. Organic pollution of groundwaters may have serious repercussions for stygofauna community structure with potentially irreversible consequences.
The relative importance of natural and anthropogenic effects on community composition of aquatic macrophytes in Mediterranean ponds
Authors: del Pozo Rocío , Fernández-Aláez Camino , Fernández-Aláez Margarita
Published: November 2018

To detect when changes in response to stressors are occurring, biomonitoring programs require an understanding of shifts in biota that occur in response to anthropogenic and natural effects. Aquatic plants are expected to reflect the environmental conditions of ponds and, according to the European Water Framework Directive, macrophytes should be considered in ecological status assessments of inland surface waters. We assessed the relative importance of natural and anthropogenic impacts on submerged, emergent and floating-leaved macrophytes in 44 ponds in Duero river basin (North Iberian Plateau). Constrained canonical ordinations included 15 taxa of submerged macrophytes and 24 species of emergent and floating-leaved macrophytes. Although the proportion of variation explained by all selected variables was relatively low (37%), we found that submerged community composition reflected the influence of natural (habitat and biotic variables) and anthropogenic effects. However, emergent and floating-leaved macrophytes were not influenced by biotic variables. Variance partitioning showed that degradation category was the best predictor of both submerged macrophytes and emergent and floating-leaved macrophyte composition. However, submerged macrophytes were more affected by chemical variables, whereas emergent and floating-leaved macrophyte composition was best explained by land-use variables. The results of this study support the use of macrophyte communities as effective indicators of the ecological status of Mediterranean ponds.
Distribution and trophic dynamics of riparian tetragnathid spiders in a large river system
Authors: Tagwireyi Paradzayi , Sullivan S. Mažeika P.
Published: November 2018

Development and agriculture are increasingly encroaching into riparian areas, with largely unknown effects on nearshore arthropods, which are important components of linked aquatic–terrestrial food webs. To assess the environmental determinants of the distribution and trophic dynamics of riparian spiders of the family Tetragnathidae, we characterised riparian habitat, collected emergent aquatic insects, and surveyed spiders in developed and rural landscapes of the Scioto River system, Ohio, USA, which provided a range of riparian land cover, nearshore vegetation types and habitat complexity. We also estimated the trophic position (TP) of Tetragnathidae and the proportion of energetic and nutritional subsidies derived from benthic algae (EBA) using naturally abundant carbon (C) and nitrogen (N) stable isotopes. Model-selection results revealed that tetragnathid spider density (1.57–3.80individualsm–1) was more sensitive to differences in overhanging vegetation than to those in aquatic food resources (i.e. emergent aquatic insects). Tetragnathidae TP, which averaged 3.16 across all 12 study reaches (range: 2.35–3.98), was largely driven by canopy density, shoreline shape, percentage overhanging vegetation and emergent-insect density. Emergent-insect density was the strongest driver of tetragnathid spider EBA (0.04–0.54, µ=0.24). Our study reinforced the notion that riparian spiders ecologically link aquatic and terrestrial ecosystems. In particular, our results further current understanding of the mechanisms affecting riparian spider distribution and trophic dynamics, particularly in the context of larger stream and river systems, given that the propensity of related research has occurred in small streams.
The Organs of Feeding and the Food of Some Australian Mullet.
Authors: Thomson JM
Published: November 2018

The morphology of the jaws of six species of mullet (Mugilidae) is
compared and illustrated. The maxilla is not fused to the skull and in two of
the species the upper end moves forward during protrusion of the mouth. The
premaxillae are protruded by the thrust from the internal hook of the
maxillae. Three adductor and two protractor muscles attach to the maxillae.
Their origins and insertions and mode of action are described. The disposition
of minute teeth on the lips, tongue, and bones of the palate and on the
pharyngeal pads is detailed. Counts of gill-rakers, pyloric caecae, and mucous
ridges in the stomach are given. The gut contents of each species as revealed
in intermittent observations are tabulated and compared with reports from
elsewhere. In general mullet feed on any small organic material wherever it
occurs in quantity, but the amount of crustacean or mollusc material is small
except in individuals with the best-developed teeth.
Further Studies on Predator Induction of Crests in Australian Daphnia
and the Effects of Crests on Predation
Authors: Barry M.J , Bayly I.A.E
Published: November 2018

The effects of notonectid predation on populations of Daphnia in a controlled seminatural environment were
studied. Log crest size was positively correlated with predator density, suggesting that crest induction was
not an all-or-nothing type of response. The size of the Daphnia population varied inversely with notonectid
density. Laboratory studies indicated that a crest-induction factor was produced by all Anisops spp. and
the one species of Enithares that were tested. Two forms of D. carinata s.1. differed in their ability to respond
to the same species of notonectid. Non-notonectid predators did not induce crests. Results of predation
experiments supported previously published indications that helmets reduce the efficiency of notonectid
predation. The effect of the helmet varied, depending on the species of notonectid and the size and form
of D. carinata s.1. The crest acted primarily to increase predator avoidance by evasion rather than by escape.
A comparative study of an ecophenotypic morph and a permanently crested morph showed that the presence
of a crest was associated with reduced fecundity. The growth rate of the permanently crested morph during
this experiment was slower, although its intrinsic mortality was considerably higher.
Root anatomy and rhizosphere ultrastructure in tropical seagrasses
Authors: Kuo J
Published: November 2018

Root morphology and histology and rhizosphere ultrastructure differ slightly among four species of
tropical seagrasses: Thalassia hemprichii (Ehrenb.) Aschers. (Hydrocharitaceae) and Cymodocea serrulata
(R. Br.) Aschers. & Magnus, Halodule uninervis (Forsk.) Aschers. and Syringodium isoetlfolium
(Aschers.) Dandy (Cymodoceaceae). Bacterial colonies have been observed in the rhizospheres of these
four seagrass species. Bacteria penetrate the epidermal cell and lyse the thick polysaccharide materials
in the walls of both epidermal and exodermal cells. Suberized lamellae in the walls of the epidermal
and exodermal cells are more resistant, but bacterial colonies eventually occur in both types of cells.
It is suggested that these rhizosphere bacterial colonies are probably not pathogenic but are mutualistic
and may be involved in nitrogen fixation and nutrient uptake by seagrass roots.
Bacterial decomposition of detritus in a tropical seagrass (Syringodium isoetifolium) ecosystem, measured with (Methyl- super-3H)thymidine
Authors: Pollard PC , Kogure K
Published: November 2018

The bacterial decomposition of detritus was measured in a tropical seagrass meadow (Syringodium
isoetifolium) off Dravuni Island, Fiji, and compared with the seagrass and algal productivity. Our
estimates of bacterial decomposition of organic matter was based on the rate at which the bacteria
divided, and this was measured by the rate at which [methyl-3H]thymidine was incorporated into the
bacterial DNA. Bacterial activity was greatest in the top 1 cm of the seagrass sediments. The number
of bacteria, their specific growth rate, and their productivity were 6.3 � 0.3 × 109 cells cm-3 , 0.04�
0.005 h-1, and 0.671 g C day-1 m-2 (depth-integrated over 30 cm), respectively.
At the sediment surface, bacterial activity coincided with the daily light intensities during the day.
Benthic microalgal production seemd to be coupled to the heterotrophic bacterial activity. However,
the amount of organic carbon passing through the bacterial decomposition process was small compared
with the total autotrophic production. The net bacterial productivity was only between 4 and 6% of
that of microalgae and 6% of that of the seagrass. The bacteria appeared to play a minor role in the
transfer of organic carbon between the primary producers and the higher trophic groups. Much of the
microalgal production could have been directly consumed by the benthic infauna, whereas the seagrass
production may have been translocated, via the seagrass roots and rhizomes, to other parts of the
plant and/or converted into more refractory or storage compounds.
Relationship between denitrification and redox potential in two sediment-water systems
Authors: Tomaszek JA
Published: November 2018

The process of denitrification was observed in vitro for two sediment-water systems. Overlying water was adjusted to a pH of 7.0, a temperature of 20�C and a nitrate concentration of 10.0 mg L-1 N-NO3-. Dissolved oxygen was maintained at a concentration above 6.0 mg L-1 O2 and incubations were carried out in the dark. These conditions insured that reduction of nitrate would result only from denitrification occurring in the sediments.
The average rate of the denitrification, calculated from the amount of nitrate removed during about one year of incubation, was 50 mg N-NO3- m-2 day-1 for lake sediments and 171 mg N-NO3- m-2 day-1 for river sediments. Continuous measurements of the redox potential at various depths in the sediments allowed determination of the zone of denitrification and estimation of its migration into the sediment. Denitrification in the sediments occurred in close proximity to the oxygenated surface of sediments.
Study of the kinetics of the denitrification process showed that the reactions for both sediment types were first-order. The exponential character of the reactions was presumably due to depletion of organic matter and could in part be a consequence of the diffusion-limited transport of nitrate into sediments. Diffusion of nitrate into the sediments was a major factor controlling denitrification rate.
Benthic microbial response to a sedimentation event at low temperature in sediments of a eutrophic lake
Authors: Tornblom E , Bostrom B
Published: November 2018

A laboratory study was performed in order to investigate the short-term response of a benthic microbial community to a spring bloom sedimentation event at low temperature. In the laboratory, organic material collected with phytoplankton nets was added to undisturbed sediment cores receiving the following four treatments: (1) addition of organic material and aeration of the water overlying the sediment, (2) only aeration, (3) only organic material addition, and (4) no organic material addition or aeration. Changes in microbial activity and biomass in the sediments were followed during six days at an experimental temperature of 4�C by means of measurements of heat production, bacterial production ([3H]thymidine incorporation), oxygen consumption, bacterial biomass, ATP concentration and chlorophyll a concentration.

In treatments with organic material additions, a response in terms of simultaneously increasing heat production, bacterial production and oxygen consumption rates was observed. Microbial activity increased slowly during the first four days in cores with organic matter addition, and between Days 4 and 6 microbial activity approximately doubled in aerated cores and decreased in non-aerated cores. Mineralization rates calculated from heat production showed that a small proportion of the added organic matter was mineralized in the surface sediment during the experiment. The results of this study show that the benthic microbial community could respond to an input of organic material at low temperature within a few days. A well oxygenated overlying water phase was necessary for the response to proceed.
Diagenetic processes and nutrient fluxes at the sediment-water interface, northern Adriatic Sea, Italy
Authors: Barbanti A , Bergamini MC , Frascari F , Miserocchi S , Ratta M , Rosso G
Published: November 2018

Early diagenetic processes that control pore-water chemistry and nutrient fluxes at the sediment-water interface were studied in late summer 1989 and spring 1990 at four sites influenced by fine sediments of the Po River. Parameters on the solid fraction (grain size and C, N and P) and pore- water profiles of SO42-, NO3-, NH3, alkalinity, PO43-, Fe, Mn, Ca and Mg were determined. Data interpretation was supported by application of kinetic and stoichiometric modelling. Nutrient fluxes were calculated by Fick's first law.
In the prodelta station the pore-water profiles showed large depletion in SO42- ions, jointly with the increase with depth of alkalinity, NH3 and PO43- concentrations, thus indicating anaerobic decomposition of organic matter, mainly due to bacterial sulfate reduction. At the other three sites the results were completely different. Nutrient concentrations in pore water were one order of magnitude lower. The peaks of alkalinity, NH3 and PO43- values near the interface and the constant presence of SO42- ions indicate aerobic decomposition of organic matter in the uppermost sediment. Nutrient fluxes showed much higher values in the prodelta station, whereas fluxes in the remaining stations decreased southward. In addition, seasonal variations produced higher summertime fluxes.

The clear-cut difference in diagenetic processes as a function of the distance from the main Po River mouth can be explained by the rapid burial of sediments rich in organic matter and inorganic compounds in the prodelta station. At the other stations, the lower sedimentation rate and more efficient bioturbation and resuspension processes cause a prolonged exposure of sediments to aerobic metabolization reactions before burial.
Phosphorus diagenesis in lake sediments: investigations using fractionation techniques
Authors: Penn MR , Auer T , Van Orman EL , Korienek JJ
Published: November 2018

A 60-cm sediment core, representing approximately 50 years of deposition, was collected from the major depositional basin of Onondaga Lake, a calcareous, hypereutrophic system in Syracuse, New York (USA). Sequential chemical extractions were performed to obtain fractional phosphorus (P) profiles, and these suggest that the sediment P available for solubilization and recycling to the water column is contained within the CaCO3-associated, Fe&Al-bound, and extractable biogenic-P fractions. Corrections were applied to account for the presence of refractory P within the CaCO3-associated and Fe&Al-bound fractions. Labile phosphorus comprises ~50% of the particulate P at the time of deposition. Reductions in labile P with depth suggest losses due to diagenesis with subsequent release to the lake water column. Rate constants for diagenesis were calculated from measurements of labile P on dated cores. Two labile components were apparent: one, exhibiting 'fast' decay (k = 4.8 year-1) was located within the upper 1 cm of sediment, and the other, characterized by 'slow' decay (k = 0.1 year-1) was located throughout the sediment to a depth of ~30 cm, below which sediment P was refractory. The ratio of the 'fast' to 'slow' fractions is approximately 1 : 1 in freshly deposited sediment. These findings facititate sediment P modelling and have bearing on the recovery of lakes following reductions in external phosphorus loads.
Risk assessment and technological options for contaminated sediments — a geochemical perspective
Authors: Forstner U
Published: November 2018

The effective long-term remediation of contaminated sediments presents a particular challenge, since the large volume and the often diverse range of contaminants in these materials makes many of the traditional remediation techniques uneconomic. Future risk assessments of contaminated sediments should focus on assessing possible chemical changes in the stored sediment (e.g. redox changes and acid production) that could lead in the long term to contaminant remobilization. New biological criteria of contaminated sediments are also needed. These risk assessments must be undertaken with the potential remediation methods in mind. Priority needs to be given to the application of new geochemical engineering techniques, including chemical stabilization by additives and storage under permanent anoxic conditions, that optimize the long-term chemical stability of contaminated sediments.
Coupled hydrological and biogeochemical model for aqueous contaminant transport
Authors: Worman A
Published: November 2018

A coupled hydrological and biogeochemical model for aqueous contaminant transport has been developed on the basis of field data on phosphorus transport in minor drainage brooks through agricultural areas in Sweden. Large scattering exists in constitutive parameters reflecting mass transfer due to solute/particle sorption and retention in bed sediments. To cope with these uncertainties, a physical/modelling framework was developed that, in a deterministic way, takes into account the most essential mechanisms controlling the transport on a regional scale and also includes randomness in process behaviour. Constitutive parameters of the governing system are conceived as stochastic, continuous fields in space and can be evaluated from field data by means of geostatistics. This modelling approach enables one to conduct analyses of uncertainty/error propagation and effects of system heterogeneity on both expected predictions and confidence intervals. Depending on the governing dimensionless numbers of the problem, changes in the covariance structure of parameter fields may cause severe deviations between statistically expected predictions associated with a stochastic parameter field and predictions based on the average value of the parameter.
A simple method to quantify sources of settling particles in lakes: Resuspension versus new sedimentation of material from planktonic production
Authors: Weyhenmeyer GA , Meili M , Pierson DC
Published: November 2018

A strong relationship (r > 0.99) between settling particulate inorganic matter (SPIM) and total settling particulate matter (SPM) was observed in 315 samples from sedimentation traps in two Swedish lakes, Lake Erken and Lake Limmaren. This relationship can be used to distinguish between different sources of SPM in pelagic systems when the inflow of allochthonous particulate matter is negligible. It is possible to quantify the proportion of material from planktonic production as well as from sediment resuspension in the flux of settling or suspended particulate matter. Furthermore, fluxes of resuspended organic particles can be quantified and distinguished from fluxes of planktonic organic particles.
Although Lake Erken and Lake Limmaren are quite different with respect to trophic level, lake surface area and water depth, the estimated proportion of resuspended particles in sedimentation traps was similar in both lakes. Resuspended particulate matter ranged from 35% of the total settling particulate matter up to 99%, with annual means of 83-94%. Moreover, for both lakes it was estimated that, even in epilimnetic traps, only 2046% (annual means) of the total organic settling particulate matter was from planktonic production, whereas 54-80% (annual means) was from sediment resuspension. This is likely to have important consequences for fluxes of nutrients and contaminants.
Tracing sources of suspended sediment in river basins: A case study of the River Culm, Devon, UK
Authors: Walling DE , Woodward JC
Published: November 2018

Information on the source of the suspended sediment transported by a river is becoming an increasingly important requirement in sediment investigations. Such information is difficult to assemble by means of traditional monitoring strategies, but the 'fingerprinting' technique offers considerable potential. The use of composite 'fingerprints' in combination with a multivariate mixing model can provide a basis for determining the relative importance of both individual areas of a catchment and specific source types. The results of applying this approach to the 276-km² basin of the River Culm in Devon, UK are presented. A suite of nine fingerprint properties was employed as a composite fingerprint, and this permitted the relative contributions of seven source types to be established. These source types represented material derived from the surface af cultivated and pasture areas on each of the three main rock types and material eroded from channel banks. By collecting samples of suspended sediment at different times during individual floods, it was possible to document changes in the relative contributions of the various sources during each flood in response to runoff source and travel times. Although the multivariate fingerprint approach has a number of limitations, it also has considerable potential as a means of tracing sources of suspended sediment within a large drainage basin.
Prediction of bottom dynamic conditions in coastal waters
Authors: Persson J , Hakanson L
Published: November 2018

Bottom dynamic conditions (areas of accumulation, erosion or transportation) in aquatic ecosystems influence the dispersal, sedimentation and recirculation of most substances, such as metals, organic toxins and nutrients. The aim of the present work was to establish a simple and general method to predict sediment types/bottom dynamic conditions in Baltic coastal areas. As a working hypothesis, it is proposed that the morphometry and the absence or presence of an archipelago outside a given coastal area regulate what factors determine the prevailing bottom dynamic conditions. Empirical data on the proportion of accumulation bottoms (BA) were collected from 38 relatively small (1-14 km²) and enclosed coastal areas in the Baltic Sea. Morphometric data were obtained by using a digital technique to transfer information from standard bathymetric maps into a computer. Data were processed by means of multivariate statistical methods. In the first model, based on data from all 38 areas, 55% of the variation in BA among the areas was statistically explained by five morphometric parameters. The data set was then divided into two parts: areas in direct connection with the open sea, and areas inside an archipelago. In the second model, based on data from 15 areas in direct connection with the open sea, 77% of the variation in BA was statistically explained by the mean depth of the deep water (the water mass below 10 m) and the mean slope. In the third model, based on data from 23 areas inside an archipelago, 70% of the variation in BA was statistically explained by the mean slope, the topographic form factor, the proportion of islands and the mean filter factor (which is a relative measure of the impact of winds and waves from outside the area). The model parameters describe the sediment trapping capacity of the areas investigated.
100 Use of Seminal Plasma, Human Chorionic Gonadotropin, and Follicular Ablation on the Interval to Follicular Wave Emergency and Dominant Follicle in Alpacas (Vicugna pacos)
Authors: Huanca W. F. , Hilari F. Y. , Villanueva J. C. , Uchuari M. , Huanca W.
Published: November 2018

Alpacas, as other camelids, are inducer ovulators and FIO/NGF-β, a protein present in the seminal plasma (SP) is reported as the responsible of the ovulation (Kershaw-Young et al. 2012 Reprod. Fertil. Dev. 24, 1093-1097, 10.1071/RD12039). However, limited and controversial information exists regarding characteristics of follicular wave in alpacas post-induction of ovulation with SP or other stimulus. The experiment was designed to determine the effect of 3 external stimulations on the interval to follicular wave emergence and the interval to dominant follicle. Adult female alpacas between 5 and 6 years old were assigned to 1 of 3 treatments: (1) SP (n = 6): 1 mL of SP IM; (2) hCG (n = 5): 1000 IU of hCG (Pregnyl, Organon-Holland, Amsterdam, the Netherlands), via IM; or (3) follicular ablation (FA, n = 6): animals were induced by ultrasound-guided ablation of the dominant follicle ≥7 mm. Alpacas from treatments 1 and 2 were examined by ultrasonography (Aloka SSD 500, transducer 7.5 MHz; Aloka, Tokyo, Japan) at 1- to 2-h intervals between 22 and 30 h after treatment or until ovulation occurred, whichever occurred first. All animals were evaluated by ultrasonography every day from Day 2 to Day 7 post-treatment and after that on Days 9, 12, and 15 post-treatment. Data from one alpaca (FA group) was excluded because of problems in the timing of ablation. Therefore, the total number of alpaca used was 16 (SP = 6, hCG = 5, and FA = 5). Results of the effect in external stimulation were analysed using ANOVA. In conclusion, interval to the emergence of a new follicular wave on the detection of follicles ≥3 mm and interval to dominant follicle ≥7 mm differed for FA compared with hCG but not compared with SP treatment.

Table 1.Follicular wave emergence (mean ± SEM) under 3 external stimulations: seminal plasma (SP), hCG, or follicular ablation (FA)
106 Effect of Heat-Stress on Antral Follicle Count and Serum Concentration of Anti-Mullerian Hormone in Bos taurus Crossbred Beef Cows
Authors: Diaz F. A. , Gutierrez E. J. , Foster B. A. , Hardin P. T. , Bondioli K. R.
Published: November 2018

Anti-mullerian hormone (AMH) has become an important tool for the selection of donors with high antral follicle count and high superovulatory response for in vivo or in vitro embryo production programs. The serum concentration of AMH is becoming a suitable selection marker because it presents minimal variation during the oestrous cycle and is repeatable during multiple cycles. It is known that heat stress results in decreased production of reproductive steroid and protein hormones in stressed animals. Importantly, the effect of heat stress on the serum concentration of AMH is still unknown. The objective of this experiment was to evaluate the antral follicle count and AMH serum concentration levels during the spring to summer transition to assess the effect of heat stress in these parameters. Ten Bos taurus crossbred non-lactating beef cows (body condition 4–8 on 9-pt scale; mean = 6.7) were used in the experiment. Dominant follicle removal was performed 5 days before sample collection and antral follicle count. Blood was collected through caudal tail venipuncture in serum collection tubes. Antral follicle count was performed through trans-vaginal ultrasound imaging of ovaries where follicles ≥2 to 3 mm were counted. Serum AMH concentration was measured utilising the Bovine AMH ELISA kit AL-114. Blood sampling and antral follicle count was performed at midmonth during April, May, June, and July. Serum AMH and antral follicle count was analysed by ANOVA. Days with average temperature-humidity index (THI) >75 (mild stress) per month were 0/30, 6/31, 26/30, and 31/31 for April, May, June, and July, respectively. Days with average THI >79 (high stress) per month were 0/30, 0/31, 6/30, and 14/31 for April, May, June, and July, respectively. The total antral follicle count for both ovaries (mean ± SE) was 36.1 ± 4.20, 48.1 ± 7.19, 34.2 ± 4.19, and 46.5 ± 5.85 for April, May, June, and July, respectively. There was no difference (P = 0.1936) in antral follicle count between months. Serum AMH concentration (ng mL−1) was 0.554 ± 0.14, 0.857 ± 0.21, 0.513 ± 0.12, and 0.575 ± 0.13 for April, May, June, and July, respectively. There was no difference (P = 0.3851) in AMH serum concentration between months. The concentration of AMH/follicle (AMH serum concentration/antral follicle count; ng mL−1) was 0.0150 ± 0.0028, 0.0166 ± 0.0018, 0.0152 ± 0.0036, and 0.0123 ± 0.0025 for April, May, June, and July, respectively. There was no difference (P = 0.7385) in the concentration of AMH/follicle between months. Results of the experiment showed that heat stress does not affect antral follicle count, AMH serum concentration, or concentration of AMH/follicle and suggest that granulosa cell function in terms of AMH production is not affected by heat stress during the spring to summer transition.
118 Identification of KISS1 in the Domestic Cat
Authors: Amelkina O. , Tanyapanyachon P. , Chatdarong K.
Published: November 2018

In the last decade, a substantial dataset has been acquired to demonstrate the critical role of kisspeptins, a family of neuropeptides derived from the KISS1 gene, in the reproduction system of mammalian and non-mammalian species. The KISS1 gene initial product can be cleaved into a 54-amino acid protein, and further degrade from the N-terminus into shorter but still active peptides (Kp-10, Kp-13, and Kp-14). The resulting peptides share a common C-terminal RF-amidated motif that leads to a strong binding with kisspeptin receptor. The Kp-10 peptide is well conserved between studied species and is thought to be essential and sufficient for the activation of kisspeptin receptor signalling pathways. In several species, the sequence of KISS1 has been obtained, and synthesised Kp-10 and Kp-54 have been successfully administered to induce ovulation. However, no studies have yet been performed to identify the KISS1 gene and its products in the domestic cat. Therefore, the aim of our work was to clone Felis catus KISS1 (fKISS1) and obtain its full sequence. Total RNA was isolated from the hypothalamus of a pubertal domestic cat collected postmortem and its quality was ensured using Agilent Bioanalyzer 2100 (RNA integrity number 8.2). Initial primers were designed based on the sequence deduced from the comparison of predicted domestic cat mRNA KISS1 sequence (XM_003999477.2) and published dog, human, cow, pig, goat, mouse, and rat mRNA KISS1 sequences. The partial mRNA fKISS1 sequence obtained here was used to design gene-specific primers for 5′ RACE and 3′ RACE PCR (GeneRacer Kit, Invitrogen, Carlsbad, CA, USA). For sequencing, 6 colonies were used per 5′ end and 3′ end of cloned fKISS1 cDNA. Additionally, the presence of KISS1 mRNA was checked via PCR in the ovaries, collected after ovariohysterectomy from domestic cats during inactive, follicular, and luteal ovarian stages. Nucleotide sequencing revealed that fKISS1 cDNA is 723 bp, and the open reading frame consists of 450 bp encoding a 149-amino acid polypeptide (kisspeptin precursor). Comparison of the overall amino acid sequence of the kisspeptin precursor revealed that domestic cat kisspeptin precursor exhibits similarity of 84, 78, 75, 68, 64, 56, and 52% to those of Hawaiian monk seal, pig, naked mole rat, human, mouse, musk shrew, and dog, respectively. The core sequence of kisspeptin, Kp-10, was highly conserved compared with other mammalian and non-mammalian species (100%, Hawaiian monk seal; 100%, mouse; 100%, pig; 100%, African clawed frog; 100%, naked mole rat; 100%, musk shrew; 90%, human; 90%, dog). Apart from hypothalamic tissue, fKISS1 has been identified in the ovaries on all stages. Obtained here results could be used for development of ovulation induction protocols in the endangered feline species. Moreover, a curious high similarity of kisspeptin sequence reported here to other induced ovulators may contribute to the research on ovulation mechanism in the domestic cat.
137 Kinetics of In Vitro Embryonic Development According to the Follicular Population in Bos Indicus
Authors: Soares J. G. , Morato F. M. , Rossi G. F. , Bayeux B. M. , Oliveira A. S. , Watanabe O. Y. , Sales J. N. S. , Lo Turco E. G. , Nogueira M. F. G. , Fontes P. K. , Watanabe Y. F. , Baruselli P. S.
Published: November 2018

The present study aimed to evaluate the effect of the follicular population from cull Nelore (Bos indicus) on the kinetics of in vitro embryonic development. At random stages of the oestrous cycle (Day –5), a total of 28 cull Nelore cows were synchronized with an intravaginal progesterone device associated with oestradiol benzoate (2.0 mg IM). At the same moment, a dose of prostaglandin F2α (2.0 mg im) was also administered to promote luteolysis and absence of corpus luteum (CL) at the time of ovum pick-up (OPU). Five days later (Day 0), all cows underwent an OPU session and the recovered oocytes were submitted to in vitro embryo production (IVEP). The same procedures were repeated 2 times at 30-day intervals (Day 25 and Day 55). Semen from a single batch of a previously tested bull was used for all IVEP. Blastocyst production and hatching were verified on Days 7, 8, and 9 of the IVEP. Data were analysed by the GLIMMIX procedure of SAS 9.3® (SAS Institute Inc., Cary, NC, USA). Data of the 3 OPU sessions were grouped and the cows were classified into 3 categories according to the follicular population: Low (19.7 ± 0.9 follicles, n = 27), Medium (33.5 ± 0.8 follicles, n = 29), and High (58.7 ± 3.2 follicles, n = 28). The Low category had a lower rate of viable oocytes [(number of viable oocytes/total number of oocytes) × 100; 62.0 v. 69.5%; P = 0.02] and cleavage rate [(number of cleaved/total number of oocytes) × 100; 55.9 v. 66.8%; P = 0.001] than the High category. The blastocyst formation rate [number of blastocysts/total number of oocytes) × 100] on Day 7 and Day 8 was lower for Low and Medium compared with the High category (Day 7: 26.1b v. 29.0b v. 35.1a %; P = 0.001; and Day 8: 29.2b v. 30.2b v. 34.7a %; P = 0.05). No differences were found in blastocysts rate on Day 9 among Low, Medium, and High categories (14.1 v. 15.9 v. 16.2%; P = 0.61). However, Low category had a lower percentage of hatched blastocysts [(number of hatched blastocysts/number of blastocysts) × 100] on Day 7 compared with High category (2.9 v. 12.0%; P = 0.01). These results reported that cull Nelore with High follicular population showed higher rates of embryo production and hatched blastocysts compared with cows with Low follicular population. We concluded that the kinetics of in vitro embryonic development was compromised in cull Nelore (Bos indicus) with low follicular population submitted to OPU-IVEP.
This research was supported by Fapesp 2012/50533–2 (GIFT).
16 Evaluation of Factors Influencing Timed Artificial Insemination in South African Communal Cows
Authors: Raphalalani Z. C. , Nedambale T. L. , Mphaphathi M. L. , Seshoka M. M. , Nkadimeng M. , Bopape M. A. , Seolwana F. L. , Mapeka M. H. , Ramukhithi F. V. , Nephawe K. A.
Published: November 2018

In South Africa, assisted reproductive technologies (ART) such as oestrus synchronization and AI in cattle have traditionally been applied in commercial production systems but not communal production systems because of several challenges such as infrastructure and cost. The study was designed to assess factors affecting response to oestrus synchronization, conception, and calving rate of organised communal cows following timed AI in Limpopo province, South Africa. A total of 140 cows were selected from organised communal villages and categorized according to body condition score (BCS), parity, age, frame size (small to medium) and breed type (Nguni, Bonsmara, and Brahman). A 9-day CIDR® (Pfizer Laboratories, New York, NY, USA) protocol was used to synchronize the selected cows. Heat mount detectors (Karma®; Four Lakes) were used to assess oestrous synchronisation responce. The AI was done twice at 36 and 48 h post-CIDR® removal using Nguni frozen–thawed semen. Pregnancy diagnosis was performed 90 days following AI using ultrasound scanner and trans-rectal palpation. Data on influence of factors such as BCS, parity, age, district, breed type, and frame size on oestrus response, conception and calving rate were analysed using logistic regression procedure of SAS. Of 140 cows synchronized, 75% (105/140) had tripped patches and underwent AI, 41% (43/105) conceived, and 36% (38/105) calved. Parity, age, breed type, and frame size did not significantly affect oestrous synchronisation response, conception, and calving rate. However, BCS significantly (P = 0.0042) affected calving rate, whereby cows in BCS of >3 had a greater probability of success than those with BCS ≤3. Small-framed Nguni and Bonsmara type cows in their first parity with a BCS ≥3 had greater odds of conceiving following timed AI. Noteworthy, calving rate in the current study was comparable to other studies under communal areas (South African Vet. Assoc. 2004 75, 30-36; Appl. Anim. Husb. Rural Develop. 2013 6, 48-54). Therefore, the current study demonstrated an opportunity to improve the production of organised communal cattle using superior sire germplasm though assisted reproductive technologies. Cows in organised communal areas have greater probability of conceiving and calving when their condition score is >3, regardless of their age, parity, size, or breed type. It is concluded, therefore, that AI technology should be applied in cows of organised communal farmers to facilitate dissemination and propagation of superior germplasm.
190 Profile of MicroRNAs of Equine Amniotic Mesenchymal Cell-Derived Microvesicles in Different Time-Span of In Vitro Culture
Authors: Lange-Consiglio A. , Capra E. , Pizzi F. , Lazzari B. , Perrini C. , Stella A. , Cremonesi F.
Published: November 2018

Mesenchymal stromal cells (MSC) were found to secrete many factors with therapeutic relevance for their antioxidants, antiapoptotic, antifibrotic, angiogenic, immunomodulatory, and chemiotactic activities. The culture supernatant of MSC (in our case, of equine amniotic-derived cells) is defined secretome or conditioned medium (CM) and it is composed of soluble and insoluble factors secreted by cells. Soluble factors are represented by cytokines and growth factors, whereas microvesicles (MV), recently demonstrated to be an integral component of cell-to-cell communication during tissue regeneration, represent insoluble factors (Bruno et al. 2009 J. Am. Soc. Nephrol. 20, 1053-1067). Our previous data showed that equine amniotic-derived CM administered in vivo in equine spontaneous tendon lesion is able to regenerate the injured tissue. overlapping the results obtained by using in vivo the cells of origin in the same pathology (Lange-Consiglio et al. 2013 Stem Cells Devel. 22, 3015-3024). We also studied the amniotic-derived MV and found that they are involved in down-regulation of pro-inflammatory genes in in vitro LPS-stressed equine tendon and endometrial cells (Lange-Consiglio et al. 2016 Stem Cells Devel. 25, 610-621; Perrini et al. 2016 Stem Cell Res. Ther. 7, 169). Usually, protocols to produce CM and MV are different: CM is collected after 48 to 96 h of culturing cells without renewal of tissue culture medium, whereas MV are collected after culturing cells overnight. In future comparative studies of regenerative medicine aiming to understand the efficacy of CM and MV, understanding the quality of secretion of cells according to culturing time is crucial. The aim of the present study was to evaluate whether different times of culture can influence the micro-RNA (miRNA) cargo of equine amniotic-derived cells and their MV. Biological triplicates of a pool of amniotic-derived cells from 2 amniotic membranes and their MV, collected after overnight or after 4 days, were used. Following miRNA extraction and library preparation, deep sequencing was carried out on Illumina HisSEqn 2000 (Illumina Inc., San Diego, CA, USA). Mirdeep2 on Illumina high-quality trimmed sequences was used to detect known miRNAs and to support the individuation of novel miRNAs. The analysis identified 335 Equus caballus miRNA, which were quantified in both amniotic cells and MV. Hierarchical clustering of 83 miRNA that were observed in all 12 samples clearly discriminated amniotic cells and MV. The profile of miRNA in AMC and MV was similar between overnight and 4 days of culture. Further functional studies on the predicted target genes and pathways involved in the biological effect of equine amniotic secretome will be performed.