Controlled reproduction in Anguilla anguilla (L.): advanced studies on broodstock management, spawning techniques and system design for artificial seed production.

The thesis aims to expose the advances achieved in the practices of captive breeding of the European eel (Anguilla anguilla). Aspects investigated concern both approaches livestock (breeding selection, response to hormonal stimulation, reproductive performance, incubation of eggs) and physiological aspects (endocrine plasma profiles of players), as well as engineering aspects. Studies conducted on various populations of wild eel have shown that the main determining factor in the selection of wild females destined to captive breeding must be the Silver Index which may determine the stage of pubertal development. The hormonal induction protocol adopted, with increasing doses of carp pituitary extract, it has proven useful to ovarian development, with a synchronization effect that is positively reflected on egg production. The studies on the effects of photoperiod show how the condition of total darkness can positively influence practices of reproductions in captivity. The effects of photoperiod were also investigated at the physiological level, observing the plasma levels of steroids ( E2, T) and thyroid hormones (T3 and T4) and the expression in the liver of vitellogenin (vtg1 and vtg2) and estradiol membrane receptor (ESR1). From the comparison between spontaneous deposition and insemination techniques through the stripping is inferred as the first ports to a better qualitative and quantitative yield in the production of eggs capable of being fertilized, also the presence of a percentage of oocytes completely transparent can be used to obtain eggs at a good rate of fertility. Finally, the design and implementation of a system for recirculating aquaculture suited to meet the needs of species-specific eel showed how to improve the reproductive results, it would be preferable to adopt low-flow and low density incubation.

Managing invasive populations of Anoplophora chinensis and A. glabripennis in Lombardy

Two Asian longhorned beetles (Coleoptera: Cerambycidae), commonly known as Citrus Longhorned Beetle (CLB), Anoplophora chinensis (Forster), and Asian Longhorned Beetle (ALB), A. glabripennis (Motschulsky), are considered the most destructive wood borers introduced in Lombardy (northern Italy). This research aimed at (1) improving laboratory rearing methods for the biological control agent Aprostocetus anoplophorae (Hym.: Eulophidae), an egg parasitoid specific to CLB, and defining release techniques allowing its establishment; (2) test the efficacy of the sentinel tree technique for the early detection of CLB; and (3) evaluating the efficacy of traps baited with artificial lures in attracting adults of ALB and possibly CLB. Several problems were faced while rearing the egg parasitoid in laboratory. It appeared that the rate of parasitism of the hosts could depend on the age of the host eggs and/or age of the laying parasitoid females. Data results from the field experiments about A. anoplophorae release-capture showed that the percentage of slits containing a CLB egg was particularly low on most sentinel trees and the percentage of CLB eggs that were killed, because of natural predators, was high. Only one egg amongst those exposed was attacked by the released parasitoid. These negative results were anyway very useful, since they provided evidence and information on the type of host plants to be used, the time necessary for the exposure of the plants to the egg-laying CLB females, the number of laying parasitoid females to be inserted per cage. The sentinel trees technique revealed to be not successful; signs and symptoms of CLB presence were not recorded during the two seasons of field observations (2012-2013). Extremely positive was instead the trial with artificial lures carried out during summer 2013. A total of 32 beetles were captured (4 ALB and 28 CLB) deploying 50 baited traps.

Influence of nest site on the behaviour of laying hens

Low rates of nest acceptance by laying hens are a major problem in commercial poultry farming operations with aviary systems, leading to costly manual collection and cleaning of mislaid eggs. To gain knowledge about factors affecting nest use, laying hens' preferences for different nest locations were tested. Nests are normally installed at one of two sites: against a wall of the hen house or integrated into one tier of the aviary rack The preferences of laying hens for different nest sites have never been examined under commercial conditions. The aim of this study is to investigate whether behavioural differences can be detected between the different nest sites. The study consists of two consecutive trials involving 5027 Lohmann Selected Leghorn hens (LSL) and 601 layer hybrids selected for extensive housing conditions (EXT). The hens were randomly assigned to eight compartments per trial in groups of 355-360 LSL or 300 EXT in a laying hen house. Four compartments were equipped with a Volito Voletage (R) aviary system (VV), and four were equipped with a Rihs Bolegell (R) aviary system (RB), both of which contained either integrated or wall-placed nests when the experiments started. A strongly balanced crossover design with four periods was used. At 36, 44 and 52 weeks of age, the nest site in four out of the eight compartments was switched. Before each change, the fronts of half of the nests were videotaped during the light period, and the behaviour throughout the main laying period was analysed. Furthermore, the numbers of nest eggs and mislaid eggs in each compartment were recorded every day. No differences in the number of mislaid eggs between the two nest sites could be detected, except at the age of 20/21 weeks when hens in VV aviaries mislaid more eggs when nests were integrated (P = 0.0012). More hens stood simultaneously in front of the integrated nests than in front of wall-placed nests (P = 0.015). Activity of the laying hens increased (P = 0.0073), and stationary behavioural patterns declined (P = 0.0093), when the nests were placed by the wall. Hens inspected integrated nests for a longer duration than wall-placed nests, but wall-placed nests were visited more frequently. In addition to the nest site, the width of the platform in front of the nest influenced laying hen behaviour. Compared with narrower platforms, balance movements decreased on wider ones. Additionally, the platform design had to be taken into account as well, given that hens could not stand or walk as securely on wooden slats as on a grid floor. (C) 2011 Elsevier B.V. All rights reserved.

Impact of copper mine tailings (stamp sand) on survival and development of aquatic organisms near Gay, Michigan

Heavy metal-rich copper mine tailings, called stamp sands, were dumped by mining companies directly into streams and along the Lake Superior shoreline, degrading Keweenaw Peninsula waterways. One of the largest disposal sites is near Gay, Michigan, where tailings have been moved along the shoreline by currents since mining ceased. As a result, the smallest sand particles have been washed into deeper water and are filling the interstitial spaces of Buffalo Reef, a critical lake trout spawning site. This research is the first to investigate if stamp sand is detrimental to survival and early development of eggs and larvae of lake sturgeon, lake trout, and Northern leopard frogs, and also examines if the presence of stamp sands influences substrate selection of earthworms. This study found that stamp sand had significantly larger mean particle sizes and irregular shapes compared to natural sand, and earthworms show a strong preference for natural substrate over any combination that included stamp sand. Additionally, copper analysis (Cu2+) of surface water over stamp sand and natural sand showed concentrations were significantly higher in stamp sand surface water (100 μg/L) compared to natural sand surface water (10 μg/L). Frog embryos had similar hatch success over both types of sand, but tadpoles reared over natural sand grew faster and had higher survival rates. Eggs of lake sturgeon showed similar hatch success and development over natural vs. stamp sand over 17 days, while lake trout eggs hatched earlier and developed faster when incubated over stamp sand, yet showed similar development over a 163 day period. Copper from stamp sand appears to impact amphibians more than fish species in this study. These results will help determine what impact stamp sand has on organisms found throughout the Keweenaw Peninsula which encounter the material at some point in their life history.

Possible open-system (hydraulic) pingos in and around the Argyre impact region of Mars

We report the observation of possible (hydraulic) open-system pingos (OSPs ) at the mid latitudes (∼37°S) in and around the Argyre impact-basin. OSPs are perennial (water)–ice cored mounds; they originate and evolve in periglacial and pro-glacial landscapes on Earth where intra- or sub-permafrost water under hydraulic/artesian pressure uplifts localised sections of surface or near-surface permafrost that then freezes in-situ. We invoke three lines of evidence in support of our analogue-based interpretation: (1) similarities of shape, size and summit traits between terrestrial OSPs and the Martian mounds; (2) clustered distribution and the slope-side location of the mounds, consistent with terrestrial permafrost-environments where OSPs are found; and, (3) spatially-associated landforms putatively indicative of periglacial and glacial processes on Mars that characterise OSP landscapes on Earth. This article presents five OSP candidate-locations and nests these mound locations within a new geological map of the Argyre impact-basin and margins. It also presents three periglacial hypotheses about the possible origin of the water required to develop the mounds. Alternative (non-periglacial) formation-hypotheses also are considered; however, we show that their robustness is not equal to that of the periglacial ones.

Antipredator defences of young are independently determined by genetic inheritance, maternal effects and own early experience in mouthbrooding cichlids.

1. Predation is a prime force of natural selection. Vulnerability to predation is typically highest early in life, hence effective antipredator defences should work already shortly after birth. Such early defences may be innate, transmitted through non-genetic parental effects or acquired by own early experience. 2. To understand potential joint effects of these sources of antipredator defences on pheno- typic expression, they should be manipulated within the same experiment. We investigated innate, parental and individual experience effects within a single experiment. Females of the African cichlid Simochromis pleurospilus were exposed to the offspring predator Ctenochromis horei or a benign species until spawning. Eggs and larvae were hand-reared, and larvae were then exposed to odour cues signalling the presence or absence of predators in a split-brood design. 3. Shortly after independence of maternal care, S. pleurospilus undergo a habitat shift from a deeper, adult habitat to a shallow juvenile habitat, a phase where young are thought to be par- ticularly exposed to predation risk. Thus, maternal effects induced by offspring predators pres- ent in the adult habitat should take effect mainly shortly after independence, whereas own experience and innate antipredator responses should shape behaviour and life history of S. pleurospilus during the later juvenile period. 4. We found that the manipulated environmental components independently affected different offspring traits. (i) Offspring of predator-exposed mothers grew faster during the first month of life and were thus larger at termination of maternal care, when the young migrate from the adult to the juvenile habitat. (ii) The offspring’s own experience shortly after hatching exerted lasting effects on predator avoidance behaviour. (iii) Finally, our results suggest that S. pleuro- spilus possess a genetically inherited ability to distinguish dangerous from benign species. 5. In S. pleurospilus, maternal effects were limited to a short but critical time window, when young undergo a niche shift. Instead, own environmental sampling of predation risk combined with an innate predisposition to correctly identify predators appears to prepare the young best for the environment, in which they grow up as juveniles.

Presence of polydnavirus transcripts in an egg-larval parasitoid and its lepidopterous host

The parasitoid Chelonus inanitus (Braconidae, Hymenoptera) oviposits into eggs of Spodoptera littoralis (Noctuidae, Lepidoptera) and, along with the egg, also injects polydnaviruses and venom, which are prerequisites for successful parasitoid development. The parasitoid larva develops within the embryonic and larval stages of the host, which enters metamorphosis precociously and arrests development in the prepupal stage. Polydnaviruses are responsible for the developmental arrest and interfere with the host's endocrine system in the last larval instar. Polydnaviruses have a segmented genome and are transmitted as a provirus integrated in the wasp's genome. Virions are only formed in female wasps and no virus replication is seen in the parasitized host. Here it is shown that very small amounts of viral transcripts were found in parasitized eggs and early larval instars of S. littoralis. Later on, transcript quantities increased and were highest in the late last larval instar for two of the three viral segments tested and in the penultimate to early last larval instar for the third segment. These are the first data on the occurrence of viral transcripts in the host of an egg-larval parasitoid and they are different from data reported for hosts of larval parasitoids, where transcript levels are already high shortly after parasitization. The analysis of three open reading frames by RT-PCR revealed viral transcripts in parasitized S. littoralis and in female pupae of C. inanitus, indicating the absence of host specificity. For one open reading frame, transcripts were also seen in male pupae, suggesting transcription from integrated viral DNA.

Effects of variation in nest curtain design on pre-laying behaviour of domestic hens

Laying hens in loose-housing systems select a nest daily in which to lay their eggs among many identical looking nests, they often prefer corner nests. We investigated whether heterogeneity in nest curtain appearance – via colours and symbols – would influence nest selection and result in an even distribution of eggs among nests. We studied pre-laying behaviour in groups of 30 LSL hens across two consecutive trials with eight groups per trial. Half of the groups had access to six identical rollaway group-nests, while the others had access to six nests of the same type differing in outer appearance. Three colours (red, green, yellow) and three black symbols (cross, circle, rectangle) were used to create three different nest curtain designs per pen. Nest position and the side of entrance to the pens were changed at 28 and 30 weeks of age, respectively, whereby the order of changes was counterbalanced across trials. Nest positions were numbered 1–6, with nest position 1 representing the nest closest to the pen entrance. Eggs were counted per nest daily from week of age 18 to 33. Nest visits were recorded individually with an RFID system for the first 5 h of light throughout weeks 24–33. Hens with access to nests differing in curtain appearance entered fewer nests daily than hens with identical nests throughout the study but both groups entered more nests with increasing age. We found no other evidence that curtain appearance affected nest choice and hens were inconsistent in their daily nest selection. A high proportion of eggs were laid in corner nests especially during the first three weeks of lay. The number of visits per egg depended upon nest position and age: it increased with age and was higher after the nest position change than before in nest position 1, whereas it stayed stable over time in nest position 6. At 24 weeks of age, gregarious nest visits (hens visiting an occupied nest when there was at least one unoccupied nest) and solitary nest visits (hens visiting an unoccupied nest when there was at least one occupied nest) accounted for a similar amount of nest visits, however, after the door switch, gregarious nest visits made up more than half of all nest visits, while the number of solitary nest visits had decreased. The visual cues were too subtle or inadequate for hens to develop individual preferences while nest position, entrance side, age and nest occupancy affected the quantity and type of nest visits.

Abundance and biomass of mesozooplankton in the Levantine Basin during the Bilim 2 cruise in April 2008

The Sesame dataset contains mesozooplankton data collected during April 2008 in the Levantine Basin (between 33.20 and 36.50 N latitude and between 30.99 and 31.008 E longitude). Mesozooplankton samples were collected by using a WP-2 closing net with 200 µm mesh size during day hours (07:00-18:00). Samples were taken from 0-50, 50-100, 100-200 m layers at 5 stations in Levantine Basin The dataset includes samples analyzed for mesozooplankton species composition, abundance and total mesozooplankton biomass. Sampling volume was estimated by multiplying the mouth area with the wire length. Sampling biomass was measured by weighing filters and then determined by sampling volume. The samples were sieved sequentially through meshes of 500 and 200 micron to separate the mesozooplankton into size fractions. The entire sample (1/2) or an aliquot of the taxon-specific mesozooplankton abundance and the total abundance of the mesozooplankton were was analyzed under the binocular microscope. Minimum 500 individuals of mesozooplankton were identified and numerated at higher taxonomic level. Taxonomic identification was done at the METU- Institute of Marine Sciences by Alexandra Gubanova,Tuba Terbiyik using the relevant taxonomic literatures. Mesozooplankton abundance and biomass were estimated by Zahit Uysal and Yesim Ak.

Ichthyoplankton density and zooplankton biomass in the Benguela upwelling system during MSM17/3 in January and February 2011

Ichthyoplankton density (fish eggs and larvae) and bulk zooplankton biomass in January/February 2011 were determined for 38 stations in the northern Benguela upwelling system, based on oblique Multinet hauls during the FS Maria S. Merian MSM17/3 cruise. A HYDROBIOS Multinet, type Midi (0.25 m**2 mouth area) was equipped with five nets of 500 µm-mesh size, temperature and oxygen probes, and an inner and outer flow meter to monitor the net's trajectory (for volume filtered calculations) as well as net clogging. The Multinet was handled over the side, towed horizontally at 2 knots. Winch speed when fearing was 0.5 or 0.3 m/s, heaving velocity 0.2 - 0.3 m/s. The Multinet was towed obliquely at 38 stations sampling the upper 200 m of the water column, which were divided into five different depth strata after inspection of temperature and oxygen concentration depth profiles. Ichthyoplankton densities and zooplankton biomass were calculated for each depth stratum (=single net) from total abundance and the volume of water filtered [individuals per m**3 and g wet weight per m**3, respectively]. In addition, densities and biomass were integrated over the area for each station [individuals per m**2], as sum of calculations for each net: Sum ([individuals per m**3]*Delta (depth bot[m]-depth top [m]).

Ichthyoplankton density and zooplankton biomass in the Benguela upwelling system during MSM19/1b in October 2011

Ichthyoplankton density (fish eggs and larvae) and bulk zooplankton biomass in October 2011 were determined for 22 stations in the northern Benguela upwelling system, based on oblique Multinet hauls during the FS Maria S. Merian MSM19/1b cruise. A HYDROBIOS Multinet, type Midi (0.25 m**2 mouth area) was equipped with five nets of 500 µm-mesh size, temperature and oxygen probes, and an inner and outer flow meter to monitor the net's trajectory (for volume filtered calculations) as well as net clogging. The Multinet was handled over the side, towed horizontally at 2 knots. Winch speed when fearing was 0.5 or 0.3 m/s, heaving velocity 0.2 - 0.3 m/s. The Multinet was towed obliquely at 22 stations sampling the upper 200 m of the water column, which were divided into five different depth strata after inspection of temperature and oxygen concentration depth profiles. Ichthyoplankton densities and zooplankton biomass were calculated for each depth stratum (=single net) from total abundance and the volume of water filtered [individuals per m**3 and g wet weight per m**3, respectively]. In addition, densities and biomass were integrated over the area for each station [individuals per m**2], as sum of calculations for each net: Sum ([individuals per m**3]*Delta (depth bot[m]-depth top [m]).

Abundance of mesozooplankton collected in October and November 1991 at 20 stations in the South Aegean Sea during O91

The O91- Mesozooplankton dataset is based on samples collected in mid October-mid November 1991 at 20 stations in the South Aegean, the SE.Ionian Sea and in NW Levantine. Samples were collected at discrete layers (from the surface till 300m. These data are published. Sampling volume was estimated by multiplying the mouth area with the wire length. The entire sample (for deep layers) or aliquot of Taxon-specific mesozooplankton abundance (1/4) (for the upper layer) was analyzed under the binocular microscope. Copepod and cladoceran species were identified and enumerated; the other zooplankters were identified and enumerated at higher taxonomic level (commonly named as zooplankton groups). Taxonomic identification was done by I.Siokou-Frangou, E.Christou, and N.Fragopoulu, using the relevant taxonomic literature. The entire sample (for deep layers) or aliquot of Mesozooplankton total abundance (1/4) (for the upper layer) was analyzed under the binocular microscope. All zooplankters were enumerated.

Diurnal egg and pellet production of Calanus finmarchicus collected during METEOR cruise M87/1 in Aril 2012

Egg and pellet production of Calanus finmarchicus was measured at 6-h intervals at all stations during the second leg of the cruise. Calanus was collected at the surface 150-m using a WP2 plankton net, and incubated in chl-max water for 24-h. Each 6 hours females were transferred to a new food solution and eggs and pellets were counted. In the end of the experiment, females were measured for prosome length. The purpose of the exercise was to calculate the minimum carbon consumption of Calanus, and how large proportion of ingestion is egested as fast sinking fecal pellets, and when.

Ichthyoplankton density and zooplankton biomass in the northern Benguela upwelling system during D356 September 2010

Ichthyoplankton density (fish eggs and larvae) and bulk zooplankton biomass in September 2010 were determined for 10 stations in the northern Benguela upwelling system, based on oblique Multinet hauls during the RRS Discovery D356 cruise. A HYDROBIOS Multinet, type Midi (0.25 m**2 mouth area) was equipped with five nets of 500 µm-mesh size, temperature and oxygen probes, and an inner and outer flow meter to monitor the net's trajectory (for volume filtered calculations) as well as net clogging. The Multinet was handled over the side, towed horizontally at 2 knots. Winch speed when fearing was 0.5 or 0.3 m/s, heaving velocity 0.2 - 0.3 m/s. The Multinet was towed obliquely at 10 stations sampling the upper 200 m of the water column, which were divided into five different depth strata after inspection of temperature and oxygen concentration depth profiles. Ichthyoplankton densities and zooplankton biomass were calculated for each depth stratum (=single net) from total abundance and the volume of water filtered [individuals per m**3 and g wet weight per m**3, respectively]. In addition, densities and biomass were integrated over the area for each station [individuals per m**2], as sum of calculations for each net: Sum ([individuals per m**3]*Delta(depth bot[m]-depth top [m]).