Marek R. Lipinski

Ethical and welfare considerations when using cephalopods as experimental animals

When using cephalopods as experimental animals, a number of factors, including morality, quality of information derived from experiments, and public perception, drives the motivation to consider welfare issues. Refinement of methods and techniques is a major step in ensuring protection of cephalopod welfare in both laboratory and field studies. To this end, existing literature that provides details of methods used in the collection, handling, maintenance, and culture of a range of cephalopods is a useful starting point when refining and justifying decisions about animal welfare. This review collates recent literature in which authors have used cephalopods as experimental animals, revealing the extent of use and diversity of cephalopod species and techniques. It also highlights several major issues when considering cephalopod welfare; how little is known about disease in cephalopods and its relationship to senescence and also how to define objective endpoints when animals are stressed or dying as a result of the experiment.

Methods for the validation of squid Age from statoliths

Growth laminations were first noted in squid statoliths by Clarke (1966), who suggested they might be useful for age determination. Spratt (1978) presented a detailed age analysis of Loligo opalescens Berry, 1911, arguing that some rings in the cephalopod statolith were deposited daily, as are fish otoliths (Panella, 1971). Growth rings in Illex illecebrosus (Lesueur, 1921) statoliths were illustrated by Lipinski (1978) with similar interpretation to that of Spratt ( op. cit. ). Several further attempts have been made to validate and/or to discuss age determination from statoliths (Hurley, Drew & Radtke, 1979; Hurley et al. 1983; Wiborg, 1979; Hurley & Beck, 1980; Kristensen, 1980; Lipinski, 1980, 1981; Rosenberg, Wiborg & Bech, 1981; Martins, 1982; Radtke, 1983; Dawe et al. 1984), and several other attempts are in preparation (R. J. Hanlon, G. V. Hurley, M. R. Clarke & R. L. Radtke, personal communications).

World squid fisheries

Abstract Some 290 species of squids comprise the order Teuthida that belongs to the molluscan Class Cephalopoda. Of these, about 30–40 squid species have substantial commercial importance around the world. Squid fisheries make a rather small contribution to world landings from capture fisheries relative to that of fish, but the proportion has increased steadily over the last decade, with some signs of recent leveling off. The present overview describes all substantial squid fisheries around the globe. The main ecological and biological features of exploited stocks, and key aspects of fisheries management are presented for each commercial species of squid worldwide. The history and fishing methods used in squid fisheries are also described. Special attention has been paid to interactions between squid fisheries and marine ecosystems including the effects of fishing gear, the role of squid in ecosystem change induced by overfishing on groundfish, and ecosystem-based fishery management.

Environmental effects on cephalopod population dynamics: implications for management of fisheries.

Cephalopods are a relatively small class of molluscs (~800 species), but they support some large industrial scale fisheries and numerous small-scale, local, artisanal fisheries. For several decades, landings of cephalopods globally have grown against a background of total finfish landings levelling off and then declining. There is now evidence that in recent years, growth in cephalopod landings has declined. The commercially exploited cephalopod species are fast-growing, short-lived ecological opportunists. Annual variability in abundance is strongly influenced by environmental variability, but the underlying causes of the links between environment and population dynamics are poorly understood. Stock assessment models have recently been developed that incorporate environmental processes that drive variability in recruitment, distribution and migration patterns. These models can be expected to improve as more, and better, data are obtained on environmental effects and as techniques for stock identification improve. A key element of future progress will be improved understanding of trophic dynamics at all phases in the cephalopod life cycle. In the meantime, there is no routine stock assessment in many targeted fisheries or in the numerous by-catch fisheries for cephalopods. There is a particular need for a precautionary approach in these cases. Assessment in many fisheries is complicated because cephalopods are ecological opportunists and stocks appear to have benefited from the reduction of key predator by overexploitation. Because of the complexities involved, ecosystem-based fisheries management integrating social, economic and ecological considerations is desirable for cephalopod fisheries. An ecological approach to management is routine in many fisheries, but to be effective, good scientific understanding of the relationships between the environment, trophic dynamics and population dynamics is essential. Fisheries and the ecosystems they depend on can only be managed by regulating the activities of the fishing industry, and this requires understanding the dynamics of the stocks they exploit.

Future challenges in cephalopod research

We thank Anto´nio M. de Frias Martins, past President of the Unitas Malacologica and Peter Marko, President of the American Malacological Society for organizing the 2013 World Congress of Malacology, and the Cephalopod International Advisory Committee for endorsing a symposium held in honour of Malcolm R. Clarke. In particular, we would like to thank the many professional staff from the University of the Azores for their hospitality, organization, troubleshooting and warm welcome to the Azores. We also thank Malcolm Clarke’s widow, Dorothy, his daughter Zoe¨, Jose´ N. Gomes-Pereira and numerous colleagues and friends of Malcolm’s from around the world for joining us at Ponta Delgada. We are grateful to Lyndsey Claro (Princeton University Press) for granting copyright permissions.

Life cycle of hake and likely management implications

Despite its economic and social importance for Namibia and South Africa, limited documented information exists regarding key aspects of the biology of deep-water hake, including its life cycle. This study utilizes data collected through the demersal surveys of the R/V Dr Fridtjof Nansen in South Africa and F/V Blue Sea 1 in Namibia to describe the migratory patterns of deep-water hake in space and time. Furthermore the study investigates aspects of the life cycle of this important species in the Benguela region. Results show that deep-water hake spawns between the western Agulhas Bank and Elands Bay in South Africa with the main nursery ground between Hondeklip Bay and the northern tip of Orange Banks. Deep-water hake in Namibia (up to the Kunene River) and along the south coast of South Africa (eastwards to Port Alfred) originate from these grounds, and undertake long-range migrations across latitudes and longitudes, respectively. This hypothesis is supported by the finding that spawning has not been observed in Namibia and there are no small juveniles along the South African south coast from the eastern border of the Agulhas Bank. The proposed pattern implies an interconnection between the Namibian and the South African components of the stock and the consequent need for a revision of the present management regime based on the assumption of stocks confined within the respective national jurisdictions. This study has used length frequency distributions in space and time in order to investigate the life cycle, in terms of origin, movement and population structure in particular, an approach that may also be useful for other widely distributed species.

Spatio‐temporal genetic structure and the effects of long‐term fishing in two partially sympatric offshore demersal fishes

Environmental gradients have been shown to disrupt gene flow in marine species, yet their influence in structuring populations at depth remains poorly understood. The Cape hakes (Merluccius paradoxus and M. capensis) are demersal species co‐occurring in the Benguela Current system, where decades of intense fishing resulted in severely depleted stocks in the past. Previous studies identified conflicting mtDNA genetic substructuring patterns and thus contrasting evolutionary trajectories for both species. Using 10 microsatellite loci, the control region of mtDNA and employing a seascape genetics approach, we investigated genetic connectivity and the impact of prolonged exploitation in the two species, which are characterized by different patterns of fishing pressure. Three consecutive years were sampled covering the entire distribution (N = 2100 fishes). Despite large estimated population sizes, both species exhibited low levels of contemporary genetic diversity (0.581 < HE < 0.692), implying that fishing has had a significant impact on their genetic composition and evolutionary trajectories. Further, for M. paradoxus, significant temporal, but not spatial, divergence points to the presence of genetic chaotic patchiness. In contrast, M. capensis exhibited a clear latitudinal cline in genetic differentiation between Namibia and South Africa (FST = 0.063, P < 0.05), with low (0.2% per generation) estimates of contemporary gene flow. Seascape analyses reveal an association with bathymetry and upwelling events, suggesting that adaptation to local environmental conditions may drive genetic differentiation in M. capensis. Importantly, our results highlight the need for temporal sampling in disentangling the complex factors that impact population divergence in marine fishes.

A new direct method of stock assessment of the loliginid squid

Hydroacoustic research conducted on chokka squid (Loligo reynaudi d’Orbigny, 1845), off the east coast of South Africa from 1994–2005, has led to the development of an innovative stock assessment technique, perhaps applicable to all loliginids that migrate inshore to spawn. This technique combines hydroacoustic biomass estimates made on the spawning concentrations inshore, and minimum biomass estimates made both inshore and offshore using demersal surveys employing the swept-area method. The hydroacoustic estimate uses an improved method to obtain target strength measurements, and squid concentrations are individually mapped from a small boat with a towed transducer. This method may be used even during intense fishing operations because of the manoeuvrability of the small boat inside a tight cluster of fishing vessels. Biomasses of the individual concentrations are then summed. The inshore biomass, also includes dispersed, mature squid migrating between concentrations, this is assessed using a concentration stability factor. The biomass of dispersed squid offshore is again calculated using the swept-area method, a well known demersal survey methodology. The biomass of concentrated (spawning) squid offshore is calculated using the same proportions between concentrated and dispersed squid which were found inshore. All four components are then summed to calculate the total biomass. The result obtained is subject to the effect of complex temporal dynamics, as new animals are recruited to the adult pool and those recently assessed migrate to other sectors of the distribution area.

Phenotypic divergence despite high gene flow in chokka squid Loligo reynaudii (Cephalopoda: Loliginidae): implications for fishery management

19 20 The commercially important chokka squid Loligo reynaudii occurring in South African 21 waters is currently managed on a single-unit stock hypothesis. We tested this assumption 22 through a spatial comparison of the morphology throughout the distributional range of the 23 species. Forty three morphometric characters were measured from 1079 chokka collected off 24 the south coast of South Africa, the west coast of South Africa, and southern Angola. While 25 no significant differences were found in the hard body parts, results from classification 26 analysis showed that though all four types of morphometric attributes (soft body parts, beaks, 27 statoliths, sucker rings) resulted in some separation, the most consistent separation of 28 samples from the three regions was based on soft body part morphometric characters. On 29 average, though dependant on the model, the overall correct classification rate ranged from 30 0.68 – 0.99 for males and 0.7 – 0.99 for females in all three regions. Previous DNA analysis 31 had revealed some genetic differences between west coast and south coast samples, 32 suggesting the confluence of the cold Benguela and warm Agulhas current may act as the 33 approximate point of a phenotypic and possible genetic breakpoint. Finer scale genetic 34 analysis of samples collected across the Benguela-Agulhas confluence reported no significant 35 genetic structuring in this area suggesting environmental heterogeneity and not restriction of 36 genetic flow/isolation as the primary driver of the observed phenotypic divergence. 37 38 INTRODUCTION 39 40 The marine environment off the coast of southern Africa is one of the most diverse, complex 41 and highly variable in the world (Lutjeharms et al., 2001). The distribution of the cape hope 42 squid Loligo reynaudii (locally known as chokka) along this coastline is largely influenced by 43 the warm Angola current and the cold Benguela current upwelling system along the West 44 African coast and the warm Agulhas current system along the south east coast (Figure 1). L. 45 reynaudii inhabits these three different environments (south coast of South Africa, west coast 46 of South Africa, and southern Angola) with an apparent break in its distribution off the coast 47 of Namibia (Shaw et al. 2010). In South Africa, two-thirds of the adult biomass is 48 concentrated on the eastern Agulhas Bank shelf where it has become an important fishery 49 resource, targeted by a major commercial hand-line jig fishery (6000–13,000t caught 50

Observations on age and reproduction of the oceanic squidAncistrocheirus lesueurii(d’Orbigny, 1842) (Cephalopoda: Ancistrocheiridae)

Despite the importance of Ancistrocheirus lesueurii in the diet of a wide variety of oceanic predators, many aspects of its biology are unknown. We report new observations on the reproductive system of the species and provide age estimates of one normal and two intersexual males based on the number of increments in the statoliths. The age of the examined mature males was estimated to be more than 2 years, increasing the maximum age known for males of the species. Female A. lesueurii have specific modified areas for spermatangia reception in the nuchal region. The morphology of the right hectocotylized ventral arm and the relatively large spermatophore are also described.