Phylogeny of the genus Aphis Linnaeus, 1758 (Homoptera: Aphididae) inferred from mitochondrial DNA sequences

Aphis is the largest aphid genus in the world and contains several of the most injurious aphid pests. It is also the most reluctant aphid genus to any comprehensive taxonomic treatment: while most species are easily classified into "species groups" that form well defined entities, numerous species within these groups are difficult to tell apart morphologically and identification keys remain ambiguous and mostly rely on host plant affiliation. In this paper, we used partial sequences of COI/COII and CytB genes to reconstruct the first phylogeny of Aphis and discuss the present systematics. The monophyly of the subgenus Bursaphis and of the tree major species groups, Black aphid, Black backed aphid and frangulae-like species was recovered by all phylogenetic analyses. However our data suggested that the nominal subgenus was not monophyletic. Relationships between major species groups were often ambiguous but "Black" and "Black backed" species groups appeared as sister clades. The most striking result of this study was that our molecular data met the same limits as the morphological characters used in classifications: mitochondrial DNA did not allow the differentiation of species that are difficult to identify. Further, interspecies relationships within groups of species for which taxonomic treatment is difficult stayed unresolved. This suggests that species delineation in the genus Aphis is often ambiguous and that diversification might have been a rapid process.     

DNA isolation of mucus from Salmo trutta (Linnaeus, 1758) and Thymallus thymallus (Linnaeus, 1758) as an alternative method to conventional fin‐clipping

The first objective of the actual project was to investigate if mucus from Salmo trutta (Linnaeus, 1758) and Thymallus thymallus (Linnaeus, 1758) is suitable for DNA extraction. In the second step, it was analyzed if the avoidance of environmentally hazardous chemicals could lead to sufficiently pure DNA isolation. Finally, it was examined if samples taken from different fish body parts yielded different grades of purity of the extracted DNA. Mucus was collected from 552 individuals of S. truttaand T. thymallusfrom 2012 to 2015. DNA was extracted with two different kits (E.Z.N.A.^® Insect DNA Kit and my‐Budget DNA Mini Kit) and two different swabs (my‐Budget roughened laminated sterile cotton swabs and common autoclaved cotton swabs). The results showed that the my‐Budget DNA Mini Kit in combination with the my‐Budget roughened laminated sterile cotton swabs was the most suitable method. It is a non‐destructive and swift technique that results in the concentration of DNA being high and pure.     

Common otolith microstructure related to key early life‐history events in flatfishes identified in the larvae and juveniles of cresthead flounder Pseudopleuronectes schrenki

Otolith microstructure of reared and wild cresthead flounder Pseudopleuronectes schrenki larvae and juveniles was used to investigate the daily periodicity of ring formation, morphological change and unique otolith structure related to important life events. By comparing microstructural features of P. schrenki with those reported for other flatfish species, it was shown that there may be microstructural features that are common to all flatfishes. In the sagittae and lapillus, a check (a distinct ring) was formed in the centre of otoliths at c. 6 days post hatching, and the daily formation of rings observed outside the check was confirmed. During metamorphosis, accessory primordia (AP) of otolith growth were formed on the outer edge of the sagittae, and the shape of the sagittae became more complex. No AP was formed on the lapilli, however, and otolith rings were concentrically formed throughout the larval and juvenile (≤51·6 mm standard length, L_S) stages. It is proposed, therefore, that lapilli are more appropriate than sagittae for analysis throughout the larval and juvenile (≤51·6 mm L_S) stages. During metamorphosis, unique rings that are relatively wide and show weak contrast are formed on lapilli (metamorphosing zone, MZ). Hence, the duration of metamorphosis, larval duration and the days of juvenile life can be estimated by the number of rings within the MZ, using rings from the check to outermost ring of the MZ, and that of rings formed outside MZ, respectively. The formation of AP on sagittae as well as the absence of AP, bilateral asymmetry and the formation of a unique structure during metamorphosis on lapilli have also been reported for other flatfishes.     

Koi herpesvirus infection in experimentally infected common carp Cyprinus carpio (Linnaeus, 1758) and three potential carrier fish species Carassius carassius (Linnaeus, 1758); Rutilus rutilus (Linnaeus, 1758); and Tinca tinca (Linnaeus, 1758) by quantitative real‐time PCR and in‐situ hybridization

Only single cells in the carrier fish species Carassius carassius (Linnaeus, 1758) for koi herpesvirus (KHV) are infected in contrast to large numbers in the susceptible species common carp Cyprinus carpio (Linnaeus 1758). Several species of the family Cyprinidae have been described as virus carrier species, showing no clinical signs of a KHV disease but able to transmit the virus to other susceptible fish. In this study, 72 common carp Cyprinus carpio (Linnaeus, 1758), 36 tench Tinca tinca (Linnaeus, 1758), 36 crucian carp Carassius carassius (Linnaeus, 1758) and 36 common roach Rutilus rutilus (Linnaeus, 1758) were experimentally infected with KHV (isolate “Israel”) by immersion and kept at 20°C. The fish were euthanized at 12 timepoints over a period of 90 days and virus DNA was quantified in tissues by a real‐time TaqMan PCR. Whereas KHV‐DNA was found in Cyprinus carpio for up to 90 days, the virus DNA was detectable only in single individuals of Rutilus rutilus, Tinca tinca and Carassius carassius for up to 25 days after experimental virus exposure. Tissue samples of Cyprinus carpio and Carassius carassius were screened by in‐situ hybridization. Positive signals were found in various organs of the common carp tested crucian carp. In the latter species a much smaller number of virus‐positive stained cells was detected compared to the infected carp.     

Testing baits prepared from by‐product of the shrimp and snow crab industry in the pot fishery for Gadus morhua (Linnaeus, 1758) and Pollachius virens (Linnaeus, 1758)

In a study between October and November 2015 in northern Norway, bait prepared from the by‐product of the shrimp and snow crab industry was employed in the pot fishery to capture cod and saithe. Frozen natural herring was used as control bait. Pots with the three bait types were placed 1,000 m apart in order to eliminate the risk of interaction between the different baits. In total 11 sampling periods were conducted. There were no significant differences in catch rates between the different baits tested on cod and saithe. The manufactured bait tested in this study showed a continuous release of odours over a 14‐day period. Results show that the water content of the bait increases rapidly in the first phase of the immersion (until day 7), and the corresponding loss of bait mass (dry weight) suggest some physical disintegration (erosion) of the bait until the water content stabilizes. This study illustrates that it is possible to replace natural herring with hydrolysates from by‐products from shrimp or snow crab industry in the Norwegian pot fishery of cod and saithe.     

Evolution of biometric and life‐history traits in lizards (Gallotia) from the Canary Islands

The aim was to study as to how biometric and life‐history traits of endemic lacertids in the Canary Islands (genus Gallotia) may have evolved, and possible factors affecting the diversification process of this taxon on successively appearing islands have been deduced. To that end, comparative analyses of sexual dimorphism and scaling of different body, head and life‐history traits to body size in 10 species/subspecies of Gallotia have been carried out. Both Felsenstein's independent contrasts and Huey and Bennett's ‘minimum evolution’ analyses show that male and female snout‐vent length (SVL) changed proportionally (sexual size dimorphism not changing with body size) throughout the evolution of these lizards and all within‐sex biometric traits have changed proportionally to SVL. Life‐history traits (size at sexual maturity, clutch size, hatchling SVL and mass, and life span) are highly correlated with adult female body size, the first two being the only traits with a positive allometry to female SVL. These results, together with the finding that the slope of hatchling SVL to female SVL regression was lower than that of SVL at maturity to female SVL, indicates that larger females reach maturity at a larger size, have larger clutches and, at the same time, have relatively smaller hatchlings than smaller females. There was no significant correlation between any pair of life‐history traits after statistically removing the effect of body size. As most traits changed proportionally to SVL, the major evolutionary change has been that of body size (a ca. threefold change between the largest and the smallest species), that is suggested to be the effect of variable ecological conditions faced by founder lizards in each island.     

Effect of environmental parameters on immune response of the Indian spiny lobster, Panulirus homarus (Linnaeus, 1758)

The high export value of the Indian spiny lobster Panulirus homarus increasingly attracts the aquaculturists for farming and fattening. However, lack of knowledge on the effect of environmental parameters on the immune system of this animal could result in high mortality, which ultimately may cause major loss to the industry. Here, we report the effect of salinity (20, 25, 35, 40, and 45 per thousand), pH (5.0, 8.0, and 9.5), dissolved oxygen (DO) (1 and 5 mg L(-1)), and ammonia-N concentration (0, 0.5, 1.5 and 3 mg L(-1)) on the immune response of P. homarus measured in the haemolymph in terms of Total Haemocyte Count (THC), phenoloxidase (PO) activity, and NBT-reduction. Our data showed significant reduction (P<0.05) in THC, and NBT-reduction at lower (20 per thousand) and higher (45 per thousand) salinities. However, PO activity showed significant disparity, showing an increasing trend from 20 to 45 per thousand. Significant reduction (P<0.05) in THC and PO activity under acidic and alkaline conditions, under hypoxic condition (1 mg L(-1)), and at the higher ammonia-N concentrations than their respective optimal conditions were observed. Thus, suggesting that extreme environmental parameters can induce modifications in the immune system of the spiny lobster P. homarus, which may enhance their susceptibility to opportunistic pathogens. The humoral parameters such as THC, PO activity, and NBT-reduction can be used as potential stress indicators for healthy management of spiny lobsters.     

Early development and allometric growth patterns in ide Leuciscus idus (Linnaeus 1758)

Allometric growth and ontogeny were studied in ide Leuciscus idus (L.) reared in laboratory conditions from 0 to 30 days post hatching (DPH), to define important steps in the species’ early life history. Based on the external morphology, eleven different stages during the early development of ide were identified (ES9_a – JS1_a; ES – embryonic stage, JS – juvenile stage). After a short period of rest, hatched individuals were visual feeders with a pelagic life. Organogenesis and allometric changes indicated that development priorities concerned feeding efficiency, respiration efficiency and locomotion performances. Also concluded was that the gradual changes observed in body proportions (which occurred across a wide range of body lengths) reflect the absence of abrupt changes in the behaviour and habitat and may also be considered as a gradual adaptation to life in flowing water.     

Life history of Coilia nasus from the Yellow Sea inferred from otolith Sr:Ca ratios

The habitat use and migratory patterns of the estuarine tapertail anchovy, Coilia nasus, from three sites near- or off-shore of the Yellow Sea, were studied by examining the environmental signatures of Sr and Ca in otoliths using electron probe microanalysis (EPMA). All the samples analyzed were offsprings of typical anadromous C. nasus. The fish grew in freshwater habitat nearly for 1?year, and then entered the brackish and sea water which could be as far as ca. 300?km from the Chinese coast line. The Sr:Ca ratios at the edge of otoliths appeared to correlated with water salinities in sampling sites.     

Factors influencing three‐spined stickleback Gasterosteus aculeatus (Linnaeus 1758) catch per unit effort

Different types of fishing gear are known to vary in catch per unit effort (CPUE), but little is known regarding this in respect to the three‐spined stickleback Gasterosteus aculeatus (Linnaeus 1758). The influence of the three‐spined stickleback CPUE by trap model, baiting and visual attractors was investigated. One trap type was found to out‐perform the other; however, baiting or attractors did not influence the CPUE. Hence, the results suggest that while the choice of trap type may have an impact on the three‐spined stickleback CPUE, baiting or attractors do not seem to improve the impact.     

Characterization of 10 microsatellite loci for the deep‐sea coral Lophelia pertusa (Linnaeus 1758)

The scleractinian coral, Lophelia pertusa, is distributed globally on continental slopes, mid‐oceanic ridges and in fjords. This species forms cold‐water reefs, which are associated with a diverse animal community. These communities are poorly understood but are currently under threat from human activities. Molecular markers are required to assess the spatial genetic population structure of this key species for management and conservation purposes. Ten polymorphic microsatellite loci were isolated for Lophelia pertusa using an enriched partial library technique. Nine loci showed significant differences from Hardy–Weinberg expected genotype frequencies, eight of which showed heterozygote deficiencies.     

Evaluating life‐history strategies of reef corals from species traits

Classifying the biological traits of organisms can test conceptual frameworks of life‐history strategies and allow for predictions of how different species may respond to environmental disturbances. We apply a trait‐based classification approach to a complex and threatened group of species, scleractinian corals. Using hierarchical clustering and random forests analyses, we identify up to four life‐history strategies that appear globally consistent across 143 species of reef corals: competitive, weedy, stress‐tolerant and generalist taxa, which are primarily separated by colony morphology, growth rate and reproductive mode. Documented shifts towards stress‐tolerant, generalist and weedy species in coral reef communities are consistent with the expected responses of these life‐history strategies. Our quantitative trait‐based approach to classifying life‐history strategies is objective, applicable to any taxa and a powerful tool that can be used to evaluate theories of community ecology and predict the impact of environmental and anthropogenic stressors on species assemblages.     

Implications of guppy (Poecilia reticulata) life‐history phenotype for mosquito control

Guppies (Poecilia reticulata) are frequently introduced to both natural and artificial water bodies as a mosquito control. Laboratory studies have demonstrated that guppies can consume large numbers of larval mosquitoes. Our study investigates how intraspecific variability in guppy phenotype affects their importance as a mosquito biocontrol and how habitat conditions (natural ponds vs. water storage containers) may influence insect biomass and guppy feeding. Using a blocked experimental design, we established stream‐side mesocosm ponds with half receiving gravel substrate to simulate pond‐bottom habitat. To provide realistic diet choices and insect abundances, we allowed the mesocosms to colonize naturally with aquatic insect larvae for 1 month before introducing guppies. We tested two distinct guppy phenotypes (from high‐ and low‐predation streams) alongside fish‐free controls. After 1 month, we measured insect biomass in the mesocosms and examined guppy gut contents to document direct predation. While overall insect biomass was not significantly different across the three fish treatments, we observed a significant reduction in mosquito biomass in fish treatments compared to fish‐free controls, as well as intraspecific differences in feeding. Overall insect biomass was significantly higher in mesocosms without gravel, while habitat condition had no effect on mosquito biomass. As guppy phenotype responds to changes in their environments, it is an important consideration for biocontrol policy to anticipate potential ecosystem effects. We close by relating our findings to other studies and by discussing the implications and potential risks of using guppies to control mosquitoes.     

Early life history of herring larvae in contrasting feeding environments determined by otolith microstructure analysis

Newly hatched autumn-spawned herring larvae Clupea harengus were released in two 2500-m³ outdoor mesocosms and reared over a 2-month period. Hydrographic conditions were similar in the two mesocosms, but the average plankton density was initially more than 10 times higher in mesocosm B compared to mesocosm A (>11⁻¹ v. <0.11⁻¹). Half-way through the experiment the feeding conditions reversed with three times higher average densities in mesocosm A than in mesocosm B (>31⁻¹ v. ∼11⁻¹). Herring larvae were sampled with a 0.3-m² two-chambered net twice weekly, and survivors were harvested by draining the mesocosms at the end of the experiment. Otolith growth trajectories of individual larvae were determined by relating radial otolith size with number of increments from the outer edge of the otolith (days before capture). The increment widths during the first 3 weeks after hatching, including the first-check size, were generally wider among larvae from mesocosm B (relatively good initial feeding conditions) than among those from mesocosm A (poor initial feeding conditions). The otolith growth pattern also confirmed that the surviving herring in mesocosm A belonged to the upper size range of larvae in the mesocosm after only 2–3 weeks from hatching; no such trend was found in mesocosm B. In both mesocosms the otolith size-at-age indicated that with the present sampling gear, herring larvae larger than 20–25 mm were underrepresented in the net samples. The information obtained from otolith-size-at-age is compared with other morphometric and biochemical measures of size and condition of larvae obtained throughout the experiment.