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ICES WKMSEL REPORT 2012 ICES ADVISORY COMMITTEE

ICES CM 2012/ACOM:59

REF. WGEF, RCM, PGMED, PGCCDBS

Report of the workshop on Sexual Maturity Staging of Elasmobranchs (WKMSEL)

11-14 December 2012

Lisbon, Portugal

International Council for the Exploration of the Sea Conseil International pour l’Exploration de la Mer

H. C. Andersens Boulevard 44–46 DK-1553 Copenhagen V Denmark Telephone (+45) 33 38 67 00 Telefax (+45) 33 93 42 15 www.ices.dk [email protected]

Recommended format for purposes of citation:

ICES. 2013. Report of the workshop on Sexual Maturity Staging of Elasmobranchs (WKMSEL) , 11-14 December 2012, Lisbon, Portugal. ICES CM 2012/ACOM:59. 66 pp.

For permission to reproduce material from this publication, please apply to the Gen-eral Secretary.

The document is a report of an Expert Group under the auspices of the International Council for the Exploration of the Sea and does not necessarily represent the views of the Council.

© 2013 International Council for the Exploration of the Sea

ICES WKMSEL REPORT 2012 i

Contents

Executive summary ................................................................................................................ 1

1 Opening of the meeting ................................................................................................ 3

1.1 Opening of the meeting and adoption of the Agenda ..................................... 3 1.2 Scientific justification and aims .......................................................................... 3

1.3 Terms of Reference ............................................................................................... 4

1.4 Data Collected before the Workshop ................................................................. 5

2 Summary of Presentations ........................................................................................... 5

2.1 Maturity scale proposed for oviparous species applied to skates from Portuguese waters ....................................................................................... 5

2.2 Studies on maturation of South-western Atlantic skates (Chondrichthyes: Rajoidei) .................................................................................. 6

2.3 MEDITS atlas on the maturity stages of Elasmobranchs ................................ 6 2.4 Validation of oviparous and viviparous ICES maturity scales

through histological analysis .............................................................................. 7

2.5 Maturity scales applied in Galeus melastomus and Squalus blainville based on the proposed scale by WKMSEL 2010 ............................. 7

2.6 Maturity staging of Chondrichthyans from south-eastern Australian .............................................................................................................. 7

2.7 Pelagic shark research program currently carried out at the Portuguese Sea and Atmospheric Institute (IPMA, I.P), with examples of applications...................................................................................... 8

2.8 Morphological analysis and description of the reproductive tract of the Ginglymostoma cirratum ......................................................................... 8

2.9 Egg cases morphometry used for specific determination of some Mediterranean skates ........................................................................................... 9

2.10 Application of the maturity scale proposed by WKMSEL 2010 for viviparous deep-water sharks ........................................................................... 10

3 The standardised macroscopic maturity scale ........................................................ 10

3.1 Oviparous maturity scales - macroscopic ....................................................... 11 3.1.1 Females .................................................................................................... 12 3.1.2 Males ........................................................................................................ 15 3.1.3 Macroscopic scales proposed for Oviparous species ........................ 17

3.2 Viviparous maturity scales - macroscopic ....................................................... 20 3.2.1 Females .................................................................................................... 20 3.2.2 Males ........................................................................................................ 21 3.2.3 Macroscopic scale proposed for Viviparous species ......................... 21

3.3 Comparison and conversion of previous maturity scales with the new one WKSMEL2 (2012) ................................................................................ 24

4 Histological determination of microscopic maturity stages ................................ 28

ii | ICES WKMSEL REPORT 2012

5 Photo reference atlas for the maturity scales proposed for oviparous and viviparous elasmobranch species ..................................................................... 32

6 Guidelines for an optimal sampling strategy for the determination of maturity stages in elasmobranchs ............................................................................ 33

7 Applications and other data collection .................................................................... 35

7.1 Introduction ......................................................................................................... 35

7.2 Demographic information ................................................................................. 35 7.3 Spatial information ............................................................................................. 37

7.4 Temporal information ........................................................................................ 37

8 Conclusion and recommendations ........................................................................... 38

9 References and working documents ........................................................................ 40

Annex 1: List of participants............................................................................................... 43

Annex 2: Agenda ................................................................................................................... 46

Annex 3: Species list with info on the study areas and mode of reproduction ................................................................................................................. 48

Annex 4: Histological photo reference for oviparous and viviparous elasmobranch species .................................................................................................. 52

Annex 5: Atlas for oviparous species ................................................................................ 53

Annex 6. Atlas for viviparous species............................................................................... 81

Annex 7: WEBGR contact to upload photo reference library ..................................... 103

Annex 8. Working documents presented by corresponding participants ............... 104

Annex 9:Maturity scales proposed by WKMSEL2 (2012) ............................................ 106

ICES WKMSEL REPORT 2012 1

Executive summary

In 2010 the first workshop on Sexual Maturity Staging of Elasmobranchs (WKMSEL) was held following the proposal by the Planning Group on Commercial Catches, Dis-cards and Biological Sampling (PGCCDBS) and the Planning group for the Mediter-ranean (PGMed 2009) in response to the requirement of the European Union to collect maturity data under the Data Collection Framework (DCF). As there was no international agreement on the maturity scales to be applied to elasmobranchs, the main objective of the workshop was to agree on a common maturity scale for elas-mobranchs (sharks, skates and rays), oviparous and viviparous species, across labora-tories. The WKMSEL reviewed the maturity scales in use and proposed two new maturity scales, one for oviparous and other for viviparous species. The WKMSEL recommended testing those scales in a wider number of species to improve the standardization before a new workshop take place. ICES requested a new meeting in 2012. The main objectives for the WKMSEL2 (2012) held in Lisbon was to review and update the maturity scales proposed in 2010, based in the experience from its applica-tion, making the descriptions simpler and easy to follow. An annotated table summa-rizing the utility and use of these maturity scales for assessment and management purposes is also presented in this report. Moreover the photographic collection built up in 2010 was updated and better reorganized thanks to the contribution of all par-ticipants.

Data

Participants from research institutes from Europe, Algeria, Australia and Brazil pro-vided their feedback on macroscopic maturity scales and microscopic assignment of maturity stages as well as on their application for both oviparous and viviparous spe-cies. Digital photos on macro and microscopic maturity stages of reproductive sys-tems were also provided.

Discussions and conclusions

The participants reviewed the maturity scales proposed by the WKMSEL in 2010 and, sepa-rating the scales by sex, produced four new macroscopic maturity scales for males and females of oviparous and viviparous species. It was proposed the adoption of these scales by all Institutes involved in elasmobranch sampling, both within the framework of EU data collection and by non-EU countries. The draft table containing a summary of the applications of maturity data and other data collection needed for stock assessment models aims to be a reference for future use of the maturity data obtained from the application of the scales, but it is expected to be reviewed by ex-perts in assessment (namely by the WGEF members) and fisheries management for further comment before being finalized. The photographic collection was updated from that of 2010, increasing the number of species and the reproductive strategies covered. Templates for illustrated maturity scales by species were also proposed for a selection of species. All participants felt that the aims of the workshop were attained and recommended the dissemination of the proposed scale to all elasmobranch scien-tists and to all laboratories involved in elasmobranch sampling. Options involving a peer-reviewing paper or a technical report from ICES or FAO were proposed, as well as the diffusion on the ICES website and on the main ICES expert groups related to this subject, namely PGCCDBS and WGEF.

2 ICES WKMSEL REPORT 2012

Abbreviations and acronyms

ARPAT – Environmental Protection Agency - Tuscany Regional ATZI – Tecnalia Sukarrieta Txatxarramendi Ugartea z/g Spain CEFAS – Centre for Environment, Fisheries and Aquaculture Science DCF – Data Collection Framework ENSSMAL Ecole Nationale Supérieure des Sciences de la Mer et de l’Amenagemet du Littoral EU – European Union F – Female FAO – Food and Agriculture Organization of the United Nations FURG – Universidade Federal do Rio Grande GFCM – General Fisheries Commission of the Mediterranean GSA – Geographical Sub Area GSI – Gonadosomatic index HCMR – Hellenic Centre for Marine Research ICCAT – International Commission for the Conservation of Atlantic Tunas ICES – International Council for the Exploration of the Sea IPMA – Instituto Português do Mar e da Atmosfera/ Portuguese Institute for the Sea and Atmosphere ISPRA – High Institute for Environmental Protection and Research M – Male MEDITS – Mediterranean International Trawl Survey NAFO – Northwest Atlantic Fisheries Organization PCL – Pre-caudal length PGCCDBS Planning Group on Commercial Catches, Discards and Biological Sampling PGMed – Planning Group for the Mediterranean RCMs – Regional Co–ordination Meetings SMALK – Sex Maturity Age Length Key TL – Total Length ToR – Terms of Reference TW – Total Weight UFRPE – Universidade Federal Rural de Pernambuco WebGR – Web services for support of Growth and Reproduction studies WGEF – Working Group on Elasmobranch Fishes WKMOG – Workshop on Maturity Ogive Estimation for Stock Assessment WKMSEL Workshop on Sexual Maturity Staging of Elasmobranchs


1 Opening of the meeting

1.1 Opening of the meeting and adoption of the Agenda

The ICES “Workshop on Sexual Maturity Staging of Elasmobranchs 2” (WKMSEL2) was hosted by the Portuguese Institute for the Sea and Atmosphere (IPMA) in Lis-bon, Portugal, from the 11 to 14 December 2012. The event was attended by 20 partic-ipants from Algeria, Australia, Brazil, Greece, Italy, Portugal and Slovenia. Several representatives from Italy, Spain, and UK participated through e-mail correspond-ence. The list of participants is attached as Annex 1.

The meeting was opened by Fabrizio Serena (co-chair of WKMSEL and WKMSEL2) with a brief overview of the background and importance of the workshop, giving reference to the importance of understanding elasmobranch reproduction from a management point of view. At the end of his presentation Fabrizio Serena, showed a final slide mentioning the concern of the corresponding participant Jim Ellis, to the need of revising the application of maturity data to the work of assessment scientists. This point was further included as a term of reference for the oviparous and vivip-arous subgroups.

Bárbara Serra-Pereira (co-chair and host) welcomed the participants, and acknowl-edged the involvement of more participants with knowledge on a wider range of species and scientific background and broadening of the workshop to more countries (from inside and outside the ICES and Mediterranean regions). Followed by a presen-tation with an overview of the main outputs from the first WKMSEL meeting (held in Malta in 2010) and also on the main goals for WKMSEL2. Bárbara Serra-Pereira also highlighted the content of the two scales proposed for oviparous and viviparous (males and females) elasmobranch species. In light of the experience of applying the scales during the previous two years, and the knowledge of the participants on addi-tional species and geographical areas, the main goals of WKMSEL2 were to revise the proposed scales and start to build an Atlas to illustrate the scales.

Following the introduction of the participants, the co-chair Bárbara Serra-Pereira proposed the approval of the Agenda (attached as Annex 2) and the adoption of three subgroups, one for the revision of the oviparous scale, another for the viviparous scales and a third group to compile the Atlas. In the end of the first day participants were assigned to each subgroup based on their experience. On behalf of Fabrizio Serena, Monica Barone was assigned as co-chair during the meeting. Catarina Maia and Joana Fernandez-Carvalho were designated as reporters for the oviparous and viviparous subgroups, respectively.

The workshop working documents and presentations (abstracts are provided on Chapter 2) are available on the ICES Sharepoint web page: https://groupnet.ices.dk/wkmsel2.

1.2 Scientific justification and aims

To understand the life cycle of a species is essential to collect reliable data about its maturity. With this information, fishery assessment scientists can provide estimates for a given species (e.g. onset of maturity by length and age, characterize the life cycle (identify recruits, juveniles, adults, pregnancy, etc.), determine the duration of egg-laying/pregnancy/gestation periods, monitor long term changes on the reproductive strategy, estimate fecundity, etc.). Furthermore, these data are relevant for many oth-er topics regarding the life cycle of any exploitable or already exploited marine living


resource, for example, energy budget allocation and maturity-survival-longevity trade offs. For that purpose, adequate maturity scales need to be produced.

Any accurate comparison of the reproductive pattern within and between species, however, would require a more standardised and consistent approach to adequately collate data from different sources. Indeed, logistic and administrative constraints, the accumulation of new knowledge or the personal opinion of the scientists in charge have determined the use of different macroscopic maturity scales, even for the same species, often in the same geographical area, in successive times. Such difficul-ties are enhanced in case of elasmobranchs species, whose reproductive patterns and their links with the life cycle traits are less documented than with the more common fish species.

The need for common maturity scales between all institutions involved in the as-sessment of fishery resources is essential for the standardization of results and com-parison of studies between laboratories that collect maturity data under the Data Collection Framework (DCF) or for any other scope. Due to the high diversity of elasmobranchs, and lack of knowledge on a large number of species, a common scale for all elasmobranchs seems adequate. Apart from a scale for each sex, the scales should also be distinct between oviparous and viviparous species, due to the major differences on the reproductive strategies of these two groups. Despite the large number of reproductive strategies of viviparous elasmobranchs available, a unique scale for viviparous should be maintained, until scientific evidence suggests other-wise. Nevertheless, the variability in reproductive mode will be taken into account when preparing the scales.

This workshop had the objectives of reviewing the previous methods and scales used for elasmobranch species, defining objective criteria to classify the maturity stages on macro scale, and reaching an agreement on the scales to be used in the future. Con-version tables to scales used in older studies will also be produced. To illustrate the scales, a stage-by-stage Atlas, with photographs provided by the different laborato-ries, on a selection of species, will be also relevant output of this workshop. Finally, it is worth pointing out the fact that if the new standard maturity scales are accepted, this will improve the standardisation and exchange of data between laboratories con-siderably. To promote the scales produced here, several strategies were discussed by the group. A digital platform is preferred, in order to associate the scale to the Atlas and allow other scientists to give their contribution.

1.3 Terms of Reference

1 ) Document the application of the new maturity scales proposed in the pre-vious workshop.

2 ) Update and review maturity information on a wide number of elasmo-branch species and reproductive strategies.

3 ) Reduce sources of error on maturity determination by validating the mac-roscopic maturity stages using histological data and biological measure-ments.

4 ) Update information on resting and regenerating stages of females and males both at the macro and micro scales and propose better descriptions.

5 ) Review and update the WKMSEL 2010 maturity scale based on the new information.


1.4 Data Collected before the Workshop

Each participant (or laboratory group) was invited to preliminary gather and present infor-mation by species and, whenever possible, separately for females and males, on:

1 ) The use of the new maturity scales proposed in the previous workshop, for both oviparous and viviparous species (i.e. difficulties, pros and cons, rec-ommendations, etc.);

2 ) Other Macroscopic maturity scales, old and in use, for both oviparous and vivip-arous species;

3 ) Digital macroscopic photos to illustrate the adopted maturity scales. The photos should be taken on fresh fish and have to include a metric reference and the sam-pling ID (for the identification of geographical area, date, bio-logical data, etc.);

4 ) Digital microscopic photos on gonads or other reproductive organs (uter-us, ovaries and oviducal glands from females and sperm ducts and semi-nal vesicles for males) to validate macroscopic observations. The photos have to include a metric reference and the sampling ID;

5 ) Protocol used for histological samples; 6 ) Methods and estimators describing maturity (e.g. measurements on repro-

ductive organs, follicle size distribution, GSI, etc.); 7 ) Reference material in order to share it among participants;

Before the workshop, each participant was invited to communicate the type of data (macro-scopic and microscopic observations, photos, maturity estimation, etc.) and the species on which they have collected information. Moreover participants were invited to communicate their intention to present a contribution during the workshop (e.g. power point presentation) and preferably to provide a working document con-taining the information collected.

2 Summary of Presentations

2.1 Maturity scale proposed for oviparous species applied to skates from Portuguese waters

Catarina Maia, Bárbara Serra-Pereira, Neide Lagarto and Ivone Figueiredo (IPMA, Portugal)

In Portugal, at IPMA, the maturity scale for oviparous species proposed in the first edition of the WKMSEL has been applied. Maturity data are collected from several skate species: thornback ray Raja clavata, blonde ray Raja brachyura, spotted ray Raja montagui, undulate ray Raja undulata, small-eyed ray Raja microocellata and cuckoo ray Leucoraja naevus. Samples are col-lected from i) IPMA research surveys along the Portuguese continental coast and ii) from commercial landings of the artisanal Portuguese fishery under the National Data Collection Program.

Since that was the first attempt to apply the proposed maturity scale and to assign the two new stages, regressing and regenerating, the main difficulties in maturity as-sessment was discussed. To this end, a photographic comparison between skate spe-cies by maturity stage was presented. The main difficulties lay in transitional stages and on assignment of regressing and regenerating individuals. It was important to note that during the sampling period there were no males observed in regressing stage, suggesting that this stage may not exist for males. Furthermore, differences in


appearance between species were also detected. As complementary information, two case studies on R. clavata and L. naevus are presented in order to illustrate the scale validation using measurements from reproductive organs. Significant differences in reproductive organs dimensions are possible to observe between different maturity stages.

2.2 Studies on maturation of South-western Atlantic skates (Chondrich-thyes: Rajoidei)

Maria Cristina Oddone and Adalto Bianchini (FURG, Brazil)

Elasmobranchs (Chondrichthyes) have an evolutionary history of more than 400 mil-lion years. In such time, they developed exquisite senses and complex reproductive modes that rival those of the most advanced tetrapods. Oviparity, the probable ances-tral condition in elasmobranchs, foreshadows the situation common in reptiles and universal in birds. The Suborder Rajoidei is oviparous and lays its egg capsules in the environment, where they develop and hatch. Oviparity in skates is single (or “exter-nal”) and all rajoids have the same general reproductive cycle. In this case, egg cap-sules are deposited in pairs with time intervals of 0-5 days between successive depositions. The assessment of chondrichthyan populations requires a quantitative approach for studying their reproductive biology, the size-at-maturity being a key parameter to be estimated. In addition, over the past thirty years, catches of rajoids have increased in the western Atlantic, mainly as a by-catch. However, sustainable catch rates in skates are actually unknown, especially in this area. The aim of the pre-sent study is to review the sexual parameters used as maturity criteria in skates for the Southwestern Atlantic Ocean studied so far. The study area was located in the southwestern Atlantic Ocean, including the continental shelves of Argentina, Uru-guay and Brazil. Traditionally, studies on skates (and elasmobranchs in general) maturation has not considered maturity stages other than immature/mature. Howev-er, in the last decade, efforts have been done in order to improve the knowledge of the skate’s maturity staging, in order to obtain more reliable maturity estimates. In this respect, our experience demonstrated that those scales that have less maturity stages (e.g., just with immature, juvenile/adolescent and mature phases, though among females we distinguish between egg-bearing and non egg-bearing females; and in the males, we recognize that within the mature there are those which storage sperm in their seminal vesicle) are more practical to use. Moreover, the more objec-tive is the scale (in terms of the terminology included) the easier it is to apply in field. A few examples of subjective terms commonly seen in elasmobranchs scales are: “uterus enlarged”, “testes small”, “oviduct widening” and so on. Attention should be paid also in the observation and record of nontraditional sexual characters in the males for maturity assessment, such as the alar thorns count and aspect and the de-velopment degree of the claspers glands. Both of them are examples of structures that can nicely aid in the discrimination between an adolescent male and a (young) ma-ture one. The microscopic assessment of sexual development and maturity onset in skates began a couple of years ago in our research group but is demonstrating to be promising in the studies in question.

2.3 MEDITS atlas on the maturity stages of Elasmobranchs

Maria Cristina Follesa and the MEDITS working group on maturity stages (Univ. of Cagliari, Italy)


A collection of the macro and micro photos of the Mediterranean elasmobranch ma-turity scales, gathered by the MEDITS working group on maturity stages, has been presented. All members group work on absolutely voluntaries basis. Each image was collected on the basis of the oviparous and viviparous maturity scales proposed dur-ing the first WKMSEL held in Malta in 2010. Presently, the collection is composed by images of 26 elasmobranchs, subdivided in 12 oviparous and 14 viviparous species. Some macro scales have been completed with also histological analysis. At the mo-ment the work is in progress but probably it will be completed within next year. The MEDITS group is open to contribute with his collection to an eventual ICES ATLAS on elasmobranch maturity stages.

2.4 Validation of oviparous and viviparous ICES maturity scales through histological analysis

Maria Cristina Follesa, M.F. Marongiu, R. Cannas, C. Porcu, A. Cau (Univ. of Caglia-ri, Italy)

One of the main Tor’s of the WKSSEL2 workshop was to perform histological anal-yses from different structures of elasmobranchs to better confirm the validation of the macro scales per-formed in the previous WKSSEL workshop. With the intent to give a contribution to this term of reference, a description of the macroscopic and micro-scopic characteristics of each stage of oviparous and viviparous scales have been pre-sented. With regard to the oviparous scales, the main features of macroscopic and histological images of the shark Galeus melastomus and the skate Dipturus oxyrin-chus have been shown. Concerning the viviparous, the characteristics of two species (Etmopterus spinax and Squalus blainville) have been shown. A suggestion to ampli-fy the description of the stage 3 (b,c,d) of viviparous scales also to the ovaries was given. This advice derives from the fact that the two species analyzed present at 3 stages a different ovaries activity: the former shows inactive gonads while the latter active ones.

2.5 Maturity scales applied in Galeus melastomus and Squalus blainville based on the proposed scale by WKMSEL 2010

Katerina Anastasopoulou and Chryssi Mytilineou (HCMR, Greece)

Different elasmobranch species were sampled during an experimental bottom long-line fishing in the Eastern Ionian Sea in deep waters from 300 to 855 m depths during two seasons within the framework of CoralFISH project. The maturity stage classifi-cation was based on the scales proposed by WKMSEL Workshop, held in Malta in 2010. At the present Workshop, macroscopic images for the oviparous Galeus me-lastomus and the viviparous Squalus blalinville were presented. The proposed mac-roscopic scale proved to be adequate when applied to female and male elasmobranchs. The main investment of the proposed maturity scale from WKMSEL was the combination of the two separated classifications systems used so far to a unique maturity scale. Problems concerning particular stages were also discussed.

2.6 Maturity staging of Chondrichthyans from south-eastern Australian

Fabian Trinnie and Terence I. Walker (DPI, Victoria, Australia)

This presentation focuses on the definitions of maturity and maternity currently used on many Australian chondrichthyan species. In many cases, chondrichthyan repro-ductive studies are undertaken through ‘at sea’ macroscopic observations but this


requires a readily observable definition of maturity. Across many Australian chon-drichthyan species, the onset of vitellogenesis (the deposition of phosvitin and lipovitellin or follicle being yolked) in females, which can be easily observed macro-scopically, has been adopted as the best indicator of ma-turity. Thus the maturity ogive describes the onset of maturity. The maturity ogive is then used in population models to determine the abundance of the mature population. However, applying the maturity ogive to recruitment rather the maternity ogive may overestimate the recruitment output, especially if the population is only reproducing biennial, trienni-ally or possibly longer. The maternity ogive which incorporates the periodicity of the reproductive cycle is a key component to fisheries models and can be defined as the proportion of females producing young into the population each year. However, to be able to determine the maternity ogive, the determination of the periodicity of the reproductive cycle is first required. Reproductive data was presented on several stingaree species: Banded stingaree Urolophus cruciatus, Sparsely-spotted stingaree U. paucimaculatus, Sandyback stingaree U. bucculentus, and the Eastern Shovelnose stingaree Trygonoptera imitata, to provide examples of varying type of reproductive cycles and to acknowledge that there is a need to unify within the literature a process required to determine the reproductive cycle e.g. all studies must collect data on the ‘period of eggs in utero’, the period of embryos in utero’, the ‘period of ovarian cy-cle’, and whether the eggs or embryos in utero grow synchronously with the ovarian cycle.

2.7 Pelagic shark research program currently carried out at the Portu-guese Sea and Atmospheric Institute (IPMA, I.P), with examples of ap-plications.

Rui Coelho, Joana Fernandez-Carvalho, Sérgio Amorim, Pedro G. Lino and Miguel Neves Santos (IPMA, Portugal)

This presentation focuses the pelagic shark research program currently carried out at the Por-tuguese Sea and Atmospheric Institute (IPMA, I.P.), in Portugal. The main research lines of this group are related to fisheries, fleet dynamics and biology of pe-lagic elasmobranchs. Within the biology component, the main research priorities in-clude life history parameters (age, growth, and reproduction), population genetics, morphometrics and habitat utilization. Most of the studies carried out in terms of re-production are focused on the estimation of parameters that can be used for popula-tion dynamics and stock assessments, and include estimations of size and age at first maturity, fecundity, sex-ratios, reproductive seasonality and periodicity. The types of data collection, including onboard sampling by fishery observers and laboratory analysis for smaller sized species, are presented and the associated difficulties dis-cussed. An example of a practical application is provided and discussed, specifically the calculation of species productivities (λ, population finite growth rate) that can be used in Ecological Risk Assessments (ERA), and that can be calculated with age-based Leslie matrices, that use life history parameters (including reproductive pa-rameters) as data inputs.

2.8 Morphological analysis and description of the reproductive tract of the Ginglymostoma cirratum

Mariana Rêgo, Fabio Hazin and Joaquim Evêncio Neto (UFRPE, Brazil)

The nurse shark is an abundant species in tropical and subtropical waters and is found along the coast of Northeast Brazil. The reproductive mode is lecithotrophic


viviparous (yolk-sac viviparity) and the periodicity of the reproductive cycle is five to six months. A total of 10 animals were collected from Mucuripe/Fortaleza, in the northeast of Brazilian and each spec-imen was measured for total length (TL) and pre-caudal length (PCL) to the nearest centimeter. The reproductive tract was weighed and measured for length and width and the reproductive condition was de-termined using the macroscopic maturity scale described by Stehmann (2002). For the histological analysis, the samples were fixed in 10% formalin (in sea water solution) for 48 hours and then transferred to 70% of ethanol. The processing and staining of tissues followed the standard protocol (Beçak and Paulete 1976 and Junqueira and Junqueira 1983). For females, the total length ranged from 1.25 to 1.50 cm. Microscop-ic analysis showed that prior to maturation, the nurse shark uterus comprises three layers: mucous, muscular and serous and the uterine lining is coated internally by a simple columnar epithelium. Just below the epithelial lining, is the lamina propria, which is undeveloped and consisting of loose connective tissue with blood and lymph vessels and showed no uterine glands. The muscle layer was composed of two well-developed layers of smooth muscle with the presence of blood and lymphatic vessels and the serous was a thin layer consisting of loose connective tissue coated externally by a simple plan epithelial tissue. The ovary showed the primary oocyte where the major part was the epigonal organ and the oocyte surrounded a layer of squamous cells of the follicle wall. In the mature ovary, the oocyte was in different stages of vitellogenesis. For males, the total length ranged from 1.50 to 2.00 m. In the immature testis, the major parts, epigonal organ, albuginea tunica and smooth mus-cle fibers were seen, and in the mature testis the connective tissue, the albuginea tuni-ca, and the beginning of spermatogonia in the somniferous tubes were seen. Also, in the mature testis, the mature spermatozoa with the Sertoli cell were seen.

2.9 Egg cases morphometry used for specific determination of some Mediterranean skates

Cecilia Mancusi, Romano Baino, Monica Barone, Fabrizio Serena (Arpat, Italy) and Gabriel Morey (Direcció General de Pesca Conselleria d'Agricultura i Pesca - Govern de les Illes Balears, Spain).

In the Mediterranean Sea at least 15 rays species have been recorded, but objective difficulties occur in the identification at specific level, especially due to a high mor-phological interspecific variability. Analyses on some morphometric parameters of several hundreds egg cases from Italy and Spain suggest a set of characteristics linked typical of some species that could be useful in the taxonomic discrimination. The species that can be determined with a significant probability from the egg-cases characteristics are, by now, Raja miraletus, Raja asterias, Raja polystigma, Raja clava-ta, Raja undulata, Raja brachyura, Dipturus nidarosiensis, Dipturus oxyrinchus and Rostroraja alba. These 9 egg cases species are described and a photo for each sample is shown. From various multivariate analyses on the recorded measurement of the egg cases, the most significant looks to be the width. Globally 558 egg cases have been examined, but, among these, only 58 egg cases where undoubtedly attributed to a known species, because found inside a pregnant female. The 495 undetermined egg cases have been later attributed to some species, with some uncertainty on the base their morphological structure. Among these, 330 specimens can be assigned to a sin-gle species: this means 67% of them. Other 11 egg cases are in the overlapping range between 3.9 and 5.6 and then can be R. undulata, R. clavata or D. oxyrinchus. Two individual egg cases of 3,8 and 4,1 are in the undetermined size range between R. polistigma and R. undulata, and so likely one of them. The last 152 specimen have


width of 3.2 and 3.3 cm: potentially thy are R. asterias or R. polistigma, but, because the area where they have found, it is most likely there are most if not all R. asterias. The methodology used is a potentially useful tool for helping in the classification of rays egg cases and individuals. Nevertheless, it is necessary to get into the analyses much a more abundant sample of known egg cases in order to define the distribution size function of each species and their probability overlapping. Despite these defi-ciencies, with the only measurement of the egg-case width is usually possible to at-tribute it to one of the most common species of rays in the Mediterranean Sea.

2.10 Application of the maturity scale proposed by WKMSEL 2010 for viviparous deep-water sharks

Teresa Moura, Neide Lagarto, Ivone Figueiredo (IPMA, Portugal)

Detail about the methodology used in the collection of the samples and the previous maturity scales adopted by IPMA were already described in the report from WKM-SEL 2010.

The new scale proposed by this working group in 2010 was adequate for many of the species of viviparous deep-water sharks and also easily adapted to our historical da-ta. However, problems were detected in the assignment of the stage 3a (“Spawning capable”) in males of the Portuguese dogfish: the maturity scale proposed mention that sharks have the testes fully segmented which might not be true for this species, where enlarged testis, not segmented or in early segmentation, were found to be “spawning capable”. Also for the males of this species, the stage 3c (“Actively spawn-ing”) was difficult to assign. The stage 4 (“Regressing”) was never recorded for any species.

In females, the definition of stage 4a (“Regressing”) mentions that the “oviducal glands diameter may be reducing”. This feature does not fit the observations for many of the species sam-pled (e.g. Portuguese dogfish and Leafscale gulper shark). Historical data shows that oviducal gland width starts to decrease right after ovula-tion, with pregnancy. Stages 4a (“Regressing”) and 4b (“Regenerating”) were usually easy to assign, especially based on the observation of the ovary and uterus. However, some occasional samples of ambiguous assignment to, , were presented to the group.

3 The standardised macroscopic maturity scale

The revision of the maturity scales collated in 2010 for oviparous and viviparous spe-cies followed the following list of tasks:

- update the list of species and study areas (also including species from geo-graphical areas outside the ICES and Mediterranean areas) (see Annex 3);

- update the histological information available for each scale (see Chapter 4);

- review the main constrains in the scale, based on the experience of using it during the last two years, and the feedback of the new participants, mainly on:

o the transition between consecutive stages;

o the description of the stages 4a (“regressing”) and 4b (“regenerating”);

o the distinction between virgin and non virgin females;

o the definitions and differences between “maturity” and “maternity” for each stage in the scale.


- propose amendments for the scale.

All the terms used to define the stages are explained in the glossary found in the re-port from 2010 WKMSEL (ICES, 2010 – Chapter 8).

The group agreed that having two separated tables for each sex might be preferable and more user-friendly. This is of special concern regarding viviparous species, since males and female reproductive cycles do not coincide (e.g. stages 3a and 3b).

The group agreed to follow the terminology proposed by ICES (Brown-Peterson et al., 2011), but it was agreed that in general the term “spawning” was not representa-tive of all elasmobranch reproductive strategies so a new term was proposed. The term “spawning” was substituted by “capable to reproduce” in stage 3a, by “egg-laying” for oviparous and “active” (adopted from previous scales) for males from the two groups in stage 4a. The term “regressing” was also changed, based on a misin-terpretation that could rise. The terms “post-laying” for oviparous and “post-partum” (adopted from previous scales) for viviparous, were used instead. In gen-eral, renaming the stages followed terminology previously used in older elasmo-branch maturity scales and standardization between oviparous and viviparous.

The group worked on simplifying the description of each stage on the scales previ-ously adopted, in order to decrease the bias between users. To aid in the use of the scales, the name of each structure was highlighted in bold, in the beginning of the description.

Regarding the assignment of the maturity stage, it was noted that it proceeds in two steps. The first step corresponds to the assignment of the maturity condition (1, 2, 3..) and the second step enters in further details (a, b, c..). Recognizing the need to have a standardized maturity stage scale and respecting the necessity to have an optional simplified version of the scale. This simplified version could be used for quick uses, but also when the capacity and experience are a constraint. The group suggested that the minimum requirement should be the assignment of the maturity/maternity condi-tion (1,2,3,..). Thus the users can decide, based on available capacity and experience, to use the detailed version of the table that will allow a better definition and compre-hension of the reproduction characteristics related to the cycle, behaviors, areas, etc. This is especially true for viviparous species, for which the maturity stage scale is more complex.

3.1 Oviparous maturity scales - macroscopic

The oviparous macroscopic scales were based on the results from skates (Rajidae and Arhyn-chobatidae) and catsharks (Scyliorhinidae), the taxa studied by the partici-pants of this work-shop (see Annex 3). Stages’ descriptions were made considering a geographic area as wide-ranging as possible. Four new species were added to the list: Dipturus nidarosiensis, Raja mi-croocellata (ICES Division IXa), Sympterygia acuta and S. bonapartei (South Brazil).

Following the recommendation from the previous workshop, the group approved the ap-plicability to other shark. Orders such as the Heterodontiformes (bullhead sharks) and Orec-tolobiformes (carpetsharks Parascycilliidae, bamboo sharks Hemiscyllidae and zebra sharks Stegostomatidae), based on the experienced from Fabian Trinnie and his colleagues in Aus-tralian waters, where these sharks mostly occur.

The main general structure of the oviparous maturity scale was maintained. The ma-turity scales for oviparous elasmobranchs include six maturity stages for females (1, 2, 3a, 3b, 4a and 4b) and five for males (1, 2, 3a, 3b and 4a). Following is a summary of


the main aspects dis-cussed when revising the maturity scales for females and males: a detailed description stage-by-stage of the changes done, recommendations to help distinguish between stages and some general recommendations in the end.

3.1.1 Females

Changes made to descriptions:

Stage 1. Immature

- The description of the ovary as “barely visible” was removed due to its ambiguity. That situation can only occur in a newly born or newly hatched fish.

- A primordium of the oviducal gland (OG) (i.e. in some species a thickening of the uteri where the gland will develop may be visible) can be identified in some skate species (e.g. R. undulata, R. clavata, Atlantoraja castelnaui, A. platana, A. cyclophora, Ri-oraja agassizi) (Fig. 3.1.1.1). The term “nidamental” was removed from the scale, adopting the terminology proposed by Hamlett (2005).

Fig. 3.1.1.1. Stage 1. Immature Raja undulata (TL= 47.5 cm) female showing the thickening of the uteri where the gland will develop (IPMA, Portugal).

Stage 2. Developing

- The term “enlarged” to describe specific condition/sizes? of the ovaries may be mis-leading, i.e. similar to stage 4a and 4b, so it was replaced by “developing”.

- Yolked follicles of different sizes may be present in the ovary, but not large ones.

Stage 3a. Capable to reproduce

Instead of referring to the presence of follicles of different sizes (very similar to the description of stage 2), the description of the ovary in this stage was simplified to “presence of large yolked follicles ready to be ovulated”.

Stage 3b. Egg-laying

Since the main feature of this stage is the presence of egg capsules, the remaining fea-tures were removed from the description.

4a.Post-laying

- The presence of large atretic follicles is the key to the assignment of this stage; post-ovulatory follicles (POFs) may also be visible (Fig. 3.1.1.2).

- The term “flaccid” was added to describe the appearance of the uteri after the egg-laying season. The uteri may also be reduced in size compared to stages 3a and 3b,


but this may be difficult to gauge when there is little knowledge on the species and no measurements are recorded.

Fig. 3.1.1.2. POF follicle observed in Leucoraja naevus (IPMA, Portugal) (left); early atretic follicle observed in Raja undulata (IPMA, Portugal) (right).

Stage 4b. Regenerating

- The reference to stage 2 was removed. The ovary may have similar follicle sizes but it is enlarged due to the previous ovarian cycle(s). No pre-ovulatory follicles are pre-sent in the ovary in this stage.

- The reduction of the OG is only detectable if measurements are taken and if there is previous knowledge of the species.

Recommendations to help to distinguish the transitions between stages:

Stage 1. Immature/Stage 2.Developing

According to Maria Cristina Oddone, a female should be assigned to the stage 1.Immature, if even when a cut is made in the ovary, no follicles are observed inside.

Stage 2. Developing/Stage 3a. Capable to reproduce

The distinction between these stages is marked by the presence of large yolked folli-cles (i.e. pre-ovulatory follicles) in 3a, apart from the development degree of OG and uterus, being the latter already fully formed. If one of these features is observed, then that female should be considered as mature (i.e. assigned to stage 3a.Capable to re-produce), since there it is likely that the female is able to contribute to reproductive output that year (Fig. 3.1.1.3).

Fig. 3.1.1.3. Stage 3a. A capable to reproduce Raja clavata (TL= 85.2 cm) (IPMA, Portugal).


Stage 3a. Capable to reproduce/Stage 4a. Post-laying

If a female has just a couple of large pre-ovulatory follicles, and few medium and small sized vitellogenic follicles (including pre-vitellogenic and vitellogenic) it is then considered as:

- 3a if the follicles are still healthy (final phase of spawning season)

or

- 4a if those follicles are entering atresia (Fig. 3.1.1.4).

Fig. 3.1.1.4. Stages 3a. Capable to reproduce and 4a. Post-laying Raja clavata (TL= 84.2 cm and 83.8 cm) (IPMA, Portugal).

Stage 4a. Post-laying/Stage 4b. Regenerating

In stage 4b there is no large degree of atresia in the ovary. The uteri and OG may ap-pear less flaccid than in 4a.post-laying female.

Stage 4b. Regenerating/Stage 2. Developing

In stage 4b, the oviducal glands and uteri are considerably enlarged, with propor-tions typical of a mature female, which are not evident in stage 2.

Stage 4b. Regenerating/Stage 3a. Capable to reproduce

In stage 4b, pre-ovulatory follicles are absent.

Other recommendations:

Ideally, the diameters of follicles should be recorded. If this is not possible, then the diameter of the maximum follicle size observed in a female could be recorded (Fitz and Daiber, 1963). A reference list of the maximum follicle size by species should be compiled, in that way it will be possible to know the pre-ovulatory size by species and help assigning females in stage 3a.Capable to reproduce. Other measurements, such as the width of the OG and uteri should also be recorded whenever possible, to allow detection of differences in size between stages, in each species. Such measure-ments provide quantitative information to support the qualitative assignment of ma-turity.

Most oviparous elasmobranch species are serial spawners, with average egg laying rates that could vary between species (e.g. maximum egg laying rate between 0.5 and 1 for Raja brachyura, R. montagui and R. clavata (Holden et al., 1971; Ellis and Shackley, 1995). Consequently, it is very subjective to detect if a female has already spawned or not. Maria Cristina Oddone showed some photos demonstrating that in some skates (e.g. Rioraja agassizi and Atlantoraja cyclophora), after egg laying, the OGs and the ante-rior uteri became very vascularized and flaccid (Fig. 3.1.1.5), but we are aware that these features may not be detectable in every female in that condition. So, for ovipa-rous species, it will not be considered the separation between females reproducing


for the first time, and those that have already contributed for recruitment in previous seasons. Although not quantified yet, the duration of a possible resting period may be of short duration for most species, i.e. less than one year (e.g. Holden 1975). It is important to highlight, that due to the reproductive strategy of oviparous elasmo-branchs, females assigned under stage 3a Capable to reproduce could be actually spawning in that period, but may have been caught in between egg produc-tion/laying episodes.

Fig. 3.1.1.5. Stage 3a.Capable to reproduce. a) Rioraja agassizi (TL= 47.4 cm); b) Atlantoraja cy-clohpora (TL= 50.0 cm) (FURG, Brazil).

In terms of assessment, oviparous females with annual cycle, after attaining maturity are expected to contribute for recruitment in every year, and should all be considered for the estimation of the Spawning Stock Biomass “SSB” (even when sampled in stag-es 4a and 4b). Although the group does not know of any example, oviparous species with longer reproductive cycles (longer than one year) may occur, and for those, the distinction between stages 4a and 4b is of particular concern, since 4b females may not be considered for SSB estimation.

3.1.2 Males

Changes made in the stages descriptions

- The term “ducts” refers to epididymis and ductus deferens.

- The level of claspers’ calcification was added to the description of stages 1.Immature, 2.Developing and 3a.Capable to reproduce, as “noncalcified”, “partially calcified” and “fully calcified” respectively.


Stage 2.Developing

The term “enlarged” may be misleading, and was replaced by “developing”.

3a.Capable to reproduce

In some species the lobules do not occupy the whole epigonal organ surface, even when fully mature (these have fully developed claspers and sperm ducts). (Fig. 3.1.2.1)

Fig. 3.1.2.1. Testes of a stage 3a.Capable to reproduce male of Atlantoraja platana (TL= 63.5 cm) (FURG, Brazil).

3b.Actively spawning

Although, sperm is not often observed flowing out of the cloaca, but inside claspers after pressing the sperm ducts the scale was maintained to follow previous descrip-tions (e.g. Stehmann, 2002).

4a.Regressing

There are not sufficient data to illustrate this stage, but it was maintained to allow future assignments.


Recommendations to help to distinguish transitions between stages:

Stage 1.Immature/Stage 2. Developing

If doubts persist when males have small claspers, undeveloped ducts (uncoiled), but some visible lobules in the testes, it is recommended to observe the presence/absence of primordial alar thorns (single row). Males with that condition (i.e. with primordial alar thorns visible, will be considered in stage 2.Developing (Fig. 3.1.2.2).

Fig. 3.1.2.2. Alar thorns of a stage 2.developing males of Atlantoraja castelnaui (left) and Sympterygia acuta (right) (FURG, Brazil).

2.Developing/3a.Capable to reproduce

Claspers and testes development are often asynchronous. But if a male has fully de-veloped testes and still flexible claspers, stage 2 should be assigned (i.e. physiological-ly it is not ready for copulating).

Other recommendations:

It is recommended to assign external and internal maturity stages, due to asynchrony of male’s reproductive organs development. But if not possible, claspers develop-ment may be used alone as maturity index, following other studies (e.g. Aasen, 1966).

3.1.3 Macroscopic scales proposed for Oviparous species

The proposed scales for oviparous elasmobranch species are presented in Tables 3.1.3.1-2.


Table 3.1.3.1. Macroscopic scale proposed for oviparous elasmobranch Females

Oviparous elasmobranchs (skates and sharks) - FEMALES

MATURITY STAGE DESCRIPTION

IMMATURE

1 IMMATURE (Immature)

Ovaries: small and whitish. Undistinguishable ovarian follicles. Oviducal gland: often not visible. In some species a thickening of the uteri where the gland will develop may be visible. Uteri: thread-like and narrow.

2 DEVELOPING (Immature)

Ovaries: follicles of different stages of development. Some small and medium sized yolked follicles may be present. Oviducal gland: distinguishable and developing. Uteri: enlarging.

MATURE

3a

CAPABLE TO REPRODUCE (mature)

Ovaries: presence of large yolked follicles ready to be ovulated. Oviducal glands: fully developed. Uteri: fully developed.

3b EGG-LAYING (mature)

Uteri: presence of egg capsules.

4a POST-LAYING (mature)

Ovaries: flaccid with few follicles of different sizes. Few large vitellogenic follicles entering atresia. POFs and atretic follicles may be visible. Oviducal glands: fully developed but may be reduced in size. Uteri: enlarged and flaccid.

4b REGENERATING (mature)

Ovaries: large with small and medium sized yolked follicles. Pre-ovulatory follicles absent. Oviducal glands: fully developed but may be reduced in size. Uteri: enlarged.


Table 3.1.3.1. Macroscopic scale proposed for oviparous elasmobranch Males

Oviparous elasmobranchs (skates and sharks) - MALES


IMMATURE

1 IMMATURE

Claspers: flexible, non-calcified and shorter than pelvic fins. Testes: small and undeveloped (in skates, sometimes with visible lobules). Ducts: straight and thread-like.

2 DEVELOPING

Claspers: flexible, partially calcified and usually as long as or longer than pelvic fins.* Testes: developing (in skates, lobules clearly visible but not fully developed). Ducts: developing and beginning to coil.

MATURE

3a

CAPABLE TO REPRODUCE

Claspers: rigid, fully calcified, and longer than pelvic fins (in some sharks they may only be as long as the pelvic fins). Testes: fully developed (in skates, with fully formed lobules). Ducts: tightly coiled and filled with sperm.

3b ACTIVE

Claspers: similar to stage 3a, however with clasper glands dilated, sometimes swollen and reddish. Sperm may be present in clasper groove or glands. Testes: similar to stage 3a. Ducts: sperm observed inside (after a cut) or flowing out of the cloaca on pressure.

4a REGRESSING

Claspers: fully formed, similar to stage 3. Testes shrunken and flaccid, (in skates, with few visible lobules). On pressure sperm does not flow. Sperm ducts: empty and flaccid

*Note: Claspers of some catsharks, such as Scyliorhinus stellaris, do not extend past the pelvic fins, even when mature.


3.2 Viviparous maturity scales - macroscopic

The maturity stage scale proposed by the WKMSEL in 2010 was reviewed by the par-ticipants, taking into account their experience in the application of the scale to a wid-er number of species. While reviewing, some issues were identified and the main points are addressed hereunder.

Considering the multitude of reproductive strategies that are found among vivip-arous elasmobranchs, and to reinforce that the scales here presented are based on a limited number of species and consequently all strategies may not be covered, the species and correspondent modes of reproduction in which these scales were based are summarize in Annex 3. The species list was updated from the last report follow-ing the new information provided by the participants. As a result ten new species were added to the list, including species from the southern West Atlantic.

3.2.1 Females

- Stage 3: The group noted that stage 3 includes both virgin (mature) and also preg-nant (maternal) (3a, 3b, 3d) females that can be confusing. It was recommended a re-classification of the numbering system of the scale (for Females only).

- Stage 3b and 3c: It was recognized that the term “segments” can create misunder-standing in the observation, thus it was suggested to simplify the description for these stages.

- Stage 3c and 3d: The group noted the subjectivity in the distinction between stages 3c and 3d. The capability for noticing this differentiation is only acquired with spe-cies-specific experience regarding state of the yolk-sac. In case of difficulties distin-guishing between these stages, taking measurements of the embryos is recommended.

- Stage 4a: The group noted that this stage should be regarded as “MATERNAL” (not just mature) condition and a possible re-definition to “Post-partum” (instead of Re-gressing) should be considered. This would be particularly important when calculat-ing the percentage of maternal females.

The group further noted that in some species (e.g. Portuguese dogfish, leafscale gulp-er shark) the oviducal glands may already be greatly reduced during pregnancy (stages 3b-3d). Therefore, on stage 4a the sentence “The oviducal gland diameter may be reducing” could be deleted from the stage definitions.

- Stage 3a to 4b “virgin issue”: The group focused on the application of the maturity stages in the calculation of reproductive parameters and comprehension of the re-productive cycle. The group discussed that females stage 3a could be interpreted ei-ther as a virgin female (maturating for the 1st time), or as a female that is mature after pregnancy and coming from stage 4b. While the current WKMSEL 2010 proposal doesn’t distinguish these two under stage 3a (virgin vs not virgin), in other classifica-tion systems those are always separated. For example, in Australia (Walker, 2005, 2007), once females get to 4b (after one cycle) it is generally recognized that it goes back to 3b, as stage 3a (under that system) is exclusively for virgin females. Those definitions will have implications for establish maturity and maternity ogives (age or length based).

The group proposed the creation of a new stage for defining females that are capable of reproducing (similar to 3a) but have gone through at least 1 cycle (non virgin). The proposed definition could be: ovaries large with large follicles, uterus enlarged with


flaccid walls, oviducal gland distinguishable. Recognizing the difficulties in the as-signment, it was agreed to not include this stage in the new scale proposed however, whenever possible to distinguish, it is recommended to record it as 3b (in the new numeration system).

As an additional note, the group observed that in aplacental species there is the risk of abortion due to stress of capture and handling, and that in those cases pregnant females (that have aborted) will probably be recorded as 4a. The group recommend-ed that when doubts arise, looking for other structures (e.g. trophonemata) could provide useful evidence for reclassifications under a pregnancy stage 3b-d.

3.2.2 Males

The descriptions of the maturity stages for viviparous elasmobranchs are very similar to oviparous ones, so those presented in section 3.1.1 were further applied to this group, with some notes described below.

- Stage 1: The group noted that some species (e.g. blue shark) never have claspers shorter than pelvic fins, thus it was suggested to rewrite the stage in order to include “usually” before “shorter”, so that the scale definitions reflect those exceptions.

-Stage 3a: The group noted that for some species (e.g. blue shark, Portuguese dogfish) the testes are enlarged and developed but not fully segmented in this stage. The group agreed to change the definition into “for some shark species testes are fully segmented” in order to explain those exceptions.

- Stage 3b: “Reddish” terminology in 3b was considered inappropriate for the species that never present this characteristic. Therefore the group agreed to delete this term from the scale.

- Stage 3a vs 3b: the group noted the difficulties when trying to distinguish between these two stages. In practice, these stages are both considered “mature” and males belonging to stage 3b might be considered a percentage of the total males at mature stage. In this regard, it was recognized that the main objective of the distinction of the stage 3b is related to the potential information on the mating seasons and copulation area. When doubts emerge, the group recommended to assign 3a in order to be more conservative.

- Stage 4: The group agreed that it is hard to distinguish the stage “regressing” for males and it was discussed if a “regressing” stage indeed exists, or if it is too short to be detected. It was considered to eliminate stage 4 for males for practical purposes, though it was further noted that the stage 4 can add valuable qualitative information in the understanding of the reproductive cycle. In addition, it was suggested that the definition of the stage 4 should include that the “seminal vesicle is developed but empty”.

As an additional note, the group discussed that for separating male’s stages, the use of the seminal vesicles is an appropriate methodology. For stage 3a, additional infor-mation that can support an active stage of copulation would be the volume of sperm in the seminal gland. Those should be relative measurements (e.g. 25%, 50%, 100%) of fullness.

3.2.3 Macroscopic scale proposed for Viviparous species

The proposed scales for viviparous elasmobranch species are presented in Tables 3.2.3.1-2.


Table 3.2.3.1. Macroscopic scale proposed for viviparous elasmobranch Females

Viviparous elasmobranchs (skates and sharks) - FEMALES

MATURITY/ MATERNITY

STAGE DESCRIPTION

IMMATURE

1 IMMATURE

Ovaries: small and whitish; undistinguishable ovarian follicles. Oviducal gland: often not visible. In some species a thickening of the uteri where the gland will develop may be visible Uteri: thread-like and narrow.

2 DEVELOPING

Ovaries: follicles of different stages of development. Some small and medium sized yolked follicles may be present. Oviducal gland: distinguishable and developing Uteri: enlarging

MATURE 3 CAPABLE to REPRODUCE

Ovaries: presence of large yolked follicles ready to be ovulated. Oviducal glands: fully developed Uteri: fully developed.

MATERNAL

4a EARLY PREGNANCY

Uteri: well filled and rounded with yolk content (usually candle shape). Embryos cannot be observed.

4b MID PREGNANCY

Uteri: well filled and rounded. Embryos are always visible, small and with a relatively large yolk sac.

4c LATE PREGNANCY

Uteri: embryos fully formed, yolk sacs reduced or absent.

5 POST-PARTUM

Ovaries: shrunken without follicle development and with atretic (degenerating) follicles. Uteri: enlarged and flaccid.

MATURE 6 REGENERATING

Ovaries: large with small and medium sized yolked follicles. Pre-ovulatory follicles absent. Atretic follicles may be present. Oviducal glands: fully developed but may be reduced in size. Uteri: enlarged.


Table 3.2.3.1. Macroscopic scale proposed for viviparous elasmobranch Males

Viviparous elasmobranchs (skates and sharks) - MALES


IMMATURE

1 IMMATURE

Claspers: flexible, non calcified and usually shorter than pelvic fins. Testes: small and undeveloped Ducts: straight and thread-like.

2 DEVELOPING

Claspers: flexible, partially calcified and as long as or longer than pelvic fins. Testes: developing and may start to segment in sharks; in rays lobules clearly visible but do not occupy the whole surface. Ducts: developing and beginning to coil.

MATURE

3a CAPABLE TO REPRODUCE

Claspers: rigid, fully calcified, longer than pelvic fins Testes: fully developed; for some shark species testes are fully segmented Ducts: tightly coiled and filled with sperm.

3b ACTIVE

Claspers: similar to stage 3a, however with clasper glands dilated, sometimes swollen. Sperm may be present in clasper groove or glands. Testes: similar to stage 3a. Ducts: sperm observed inside after a cut or flowing out of the cloaca on pressure.

4 REGRESSING

Claspers: fully formed, similar to stage 3. Testes shrunken and flaccid, (in skates, with few visible lobules). On pressure sperm does not flow. Sperm ducts: empty and flaccid. Seminal vesicle developed but empty


3.3 Comparison and conversion of previous maturity scales with the new one WKSMEL2 (2012)

The terminology commonly used to describe different reproductive phases in elas-mobranchs is variable among authors. A broad comparison between the reproductive terminologies from several published works on oviparous elasmobranchs throughout the world can be consulted in Serra-Pereira et al. (2011). Following is a comparative summary of the terminologies from old and new scales used by WKMSEL and WKMSEL2 participants and from literature (Tables 3.3.1 to 3.3.4)

Table 3.3.1. Conversion table between previous maturity scales and the WKMSEL 2 scale for Mac-roscopic scale proposed for Female oviparous elasmobranchs

Holden and Raitt

(1974)

Relini et al (1999) ”OLD

MEDITS”

Steh-mann (2002)

Relini et al (2008) “MEDIT

S Kavala”

Ungaro (2008)

“ICES”

ICES (2009)

“WGEF”

WKMSEL

2010

Serra-Pereira

et al. (2011)

WKMSEL2

(2012)

1

Imma-ture

1

Imma-ture

A or 1

Imma-ture,

juvenile

1

Imma-ture/

Virgin

F1

Imma-ture,

Juvenile

A

Imma-ture

1

Imma-ture

1

Imma-ture

1

Immature

2

Matur-ing

2

Matur-ing

2 or B

Matur-ing,

adoles-cent

2

Matur-ing

F2

Adoles-cent,

Matur-ing

B

Matur-ing

2

Develop-ing

2

Develop-ing

2

Develop-ing

3

Mature

3

Mature

3 or C

Mature, adult

3a

Mature

F3

Adult, Mature

C

Mature

3a

Spawn-ing ca-pable

3a

Spawn-ing ca-pable

3a

Capable to reproduce

3

Mature

3

Mature

D or 4

Active

E or 5

Ad-vanced

F or 6

Extrud-ing

3b

Mature/

Extrud-ing,

Active

F4

Active, uterine stage; Ad-

vanced, uterine stage;

Extrud-ing,

uterine stage

D

Active

3b

Actively spawn-

ing

3b

Actively spawn-

ing

3b

Egg-laying

No stage compari-

son

No stage compari-

son

No stage compari-

son

4a

Resting

No stage compari-

son

No stage compar-

ison

4a

Regress-ing

4

Regress-ing

4

Post-laying

No stage compari-

son

No stage compari-

son

No stage compari-

son

4

Resting

No stage compari-

son

No stage compar-

ison

4b

Regen-erating

5

Regen-erating

5

Regenerat-ing


Table 3.3.2. Conversion table between previous maturity scales and the WKMSEL 2 scale for Mac-roscopic scale proposed for Male oviparous elasmobranchs

Holden and Raitt (1974)

Relini et al (1999) ”OLD MEDITS”

Stehmann (2002)

Relini et al (2008) “MEDITS Kavala”

Ungaro (2008) “ICES”

ICES (2009) “WGEF”

WKMSEL 2010

Serra-Pereira et al. (2011)

WKMSEL2 (2012)

1 Immature

1 Immature

A or 1 Immature, juvenile

1 Immature/ Virgin

M1 Juvenile

A Immature

1 Immature

1 Immature

1 Immature

2 Maturing

2 Maturing

2 or B Maturing, adolescent, sub-adult

2 Maturing

M2 Adolescent, Maturing

B Maturing

2 Developing

2 Developing

2 Developing

3 Mature

3 Mature

3 or C Mature, adult

3a Mature

M3 Adult, Mature

C Mature

3a Spawning capable

3a Spawning capable

3a Capable to reproduce

3 Mature

3 Mature

4 or D Active

3b Mature/ Extruding, Active

M4 Active, copulating

D Active

3b Actively spawning


3b Active

No stage comparison

No stage comparison

No stage comparison

4a Resting

No stage comparison

No stage comparison

4 Regressing

4 Regressing

4 Regressing


Table 3.3.3. Conversion table between previous maturity scales and the WKMSEL 2 scale for Macroscopic scale proposed for Female viviparous elasmobranchs

Stehmann (2002) Stehmann (2002) Adapted by Samira

Walker (2005)




Figueiredo et al. (2008) (Stehmann, 2002 adaptation)

WKMSEL (2010)

WKMSEL2 (2012)

A or 1 Immature, juvenile

A Immature, juvenile

U=1 Immature

1 Immature/ Virgin

F1 Immature, Juvenile

A Immature

1 Juvenile

1 Immature

1 Immature

2 or B Maturing, adolescent

B Maturing, adolescent, sub-adult

U=2 Maturing

2 Maturing

F2 Adolescent, Maturing

B Maturing

2 Maturing

2 Developing

2 Developing


C Mature, adult

U=3 Mature 3a, 3b

F3 Adult, Mature

C Mature

3 Adult

3a Capable to reproduce

3 Capable to reproduce

4 or D Developing

C1 Active

U=4 Mature Eggs in utero only

4a F4 D Early gravid

4 Developing

3b Early pregnancy

4a Early pregnancy

E or 5 Differentiating

C2

U=5 Mature Embryos in utero

4b F5 E Mid-term gravid

5 Differentiation

3c Mid pregnancy

4b Mid pregnancy

F or 6 Expecting

C2

U=5 Mature Embryos in utero

5 F6 F Late gravid

6 Extrusion

3d Late pregnancy

4c Late pregnancy

G or 7 Post-natal, spent C3

U=6 Mature Post-partum

6 F7 G Post-partum

7 Resting

4a Regressing

5 Post-partum

B or 2 (Maturing for the 2nd or more times)

No stage comparison

U=6 Mature Post-partum

No stage comparison

No stage comparison

No stage comparison

8 Maturing (not for the first time)

4b Regenerating

6 Regenerating


Table 3.3.4. Conversion table between previous maturity scales and the WKMSEL 2 scale for Macroscopic scale proposed for Male viviparous elasmobranchs

Stehmann (2002) Stehmann (2002)

Adapted by Sami-ra

Walker (2005)




Figueiredo et al. (2008)

(Stehmann, 2002 adaptation)

WKMSEL (2010)

WKMSEL (2012)

A or 1 Immature, juve-

nile

A Immature, juve-

nile

GI=1 VI=1 CI=0

Immature

1 Immature/

Virgin

M1 Juvenile

A Immature

1 Juvenile

1 Immature

1 Immature

2 or B Maturing, adoles-

cent, sub-adult

B Maturing, adoles-

cent, sub-adult

GI=2 VI=not defined

CI=1 Immature

2 Maturing

M2 Adolescent, Ma-

turing

B Maturing

2 Maturing

2 Developing

2 Developing


C Mature, adult

GI=3 VI=2 CI=2

Mature

3a Mature

M3 Adult, Mature

C Mature

3 Adult

3a Spawning capable

3a Capable to repro-

duce

4 or D Active

C1 Active

GI=3 VI=2 CI=2

Mature 3b

Mature/ Extruding, Active

M4 Active,

copulating

D Active

4 Active


3b Active

No stage compar-ison

No stage compari-son

GI=3 VI=3 CI=2

Mature





4 Regressing

4 Regressing


4 Histological determination of microscopic maturity stages

The group acknowledges that, although not used as a routine procedure for maturity stage assignment on elasmobranch species, histology could be used as a validation tool when doubts persist assigning the macroscopic stages. For future references, it is recommended that if new information on maturity stages validation using histology is collected, the results should be documented and presented in the next WKMSEL meeting. In light of that new information the scales should be revised accordingly.

The microscales proposed by the WKMSEL aimed to be as wide-ranging as possible, despite the information available being limited to just a few species. Following the scale proposed in the previous workshop, some structural changes in the description of the different stages were made in both males and females microscopic scales. De-scriptions were structured by reproductive organ (highlighted in bold format) in or-der to improve tables reading. Some changes were also made on the text. Some participants provided updates to the histological photo reference, and their contribu-tions are presented on Annex 4.

The microscopic maturity scales for females and males for both oviparous and vivip-arous species are presented below (Tables 4.1, 4.2 and 4.3).

FEMALES

In light of the new information available, following is a brief description of some of the changes made to the scale’ content:

- Stage 2: given that vitellogenesis begins during this stage the following addition was proposed: “Vitellogenic follicles can be present”.

- Stage 3a: regarding that a female “capable to reproduce” may already have released some egg capsules, the sentence concerning the possible observation of post-ovulatory follicles was added.

- Stage 3b: A sentence describing that ovaries and uteri are similar to stage 3a was added.

- Stage 4a: Once that there are no observations on the production of secretions in the uteri during stage 4a, this reference was removed.

- Observations on stage 4b are not available.

MALES

No additional information on males was available.


Table 4.1. Microscopic maturity scale for female oviparous elasmobranch

STAGE FEMALES – Microscopic scale - Oviparous -

1

Ovaries: early pre-vitellogenic follicles, connected to the germinal epithelium and tunica albuginea. Uteri: composed mainly by connective tissue, covered by simple columnar epithelium with some invaginations.

2

Ovaries: pre-vitellogenic follicles of various sizes. Vitellogenic follicles can be present. Oviducal glands: gland tubules beginning to form (early stage) or completely formed with secreting zones differentiated (late stage). Secretions may be observed inside the gland tubules. Uteri: epithelium more invaginated and vascularized than in immature females. Secretions may be observed inside secretory cells.

3a

Ovaries: follicles in all stages of development, being vitellogenic follicles the most abundant. Post-ovulatory follicles can be present. Oviducal glands: filled with secretions in the tubules. Uteri: highly invaginated, with longitudinal folds producing secretions to the lumen. Stratified epithelium.

3b

Ovaries: follicles similar to 3a. Post-ovulatory follicles can be present. Oviducal glands: tubules full of secretion materials. Secretions also present in the gland lumen. Uteri: structure similar to 3a.

4a

Ovaries: follicles in all stages. Post-ovulatory and atretic follicles, in which the basal lamina appears collapsed and invade the central lumen, are found. Oviducal glands: fully formed but without secretions in the lumen. Uteri: fully formed.

4b

No observations available


Table 4.2. Microscopic maturity scale for female viviparous elasmobranch

STAGE FEMALES – Microscopic scale - Viviparous -

1

Ovaries: early pre-vitellogenic follicles, connected to the germinal epithelium and tunica albuginea. Uteri: same as in oviparous

2

Ovaries: pre-vitellogenic follicles of various sizes. Vitellogenic follicles can be present. Oviducal glands: no observations available. Uteri: no observations available.

3 No observations available

4a No observations available

4b No observations available

4c No observations available




Table 4.3. Microscopic maturity scale for oviparous and Male viviparous elasmobranch

STAGE MALES - Microscopic scale – Oviparous and Viviparous -

1

Testis: gonocytes, spermatogonia (more abundant), primary and secondary spermatocytes embedded in the epigonal organ. More advanced stages of spermatocysts are rarely observed (spermatids and spermatozoa). Sperm ducts: no observations available.

2

Testis: spermatocysts in all spermatogenesis stages, though generally, secondary spermatocytes are the most abundant. Spermatids and spermatozoa are more abundant than in immature males. Sperm ducts: start to differentiate in villosities. Spermatozoa absent.

3a

Testis: spematocysts in all stages. Spermatids and spermatozoa are more abundant than in stage 2. Sperm ducts: full with seminal liquid, denser in the deferent ducts. Spermatozoa bundles are observed in the lumen.

3b

Testis: description similar to 3a. Spermatozoa are the most abundant stage observed. Spermatozoa heads are arranged to the periphery while the tails are spiralled towards the lumen. Sperm ducts: description similar to 3a.

4a

Testis: lobules previously containing sperm, appear empty. It is possible to find clusters of sperm. Sperm ducts: no observations available.


5 Photo reference atlas for the maturity scales proposed for oviparous and viviparous elasmobranch species

In the ICES area and the Mediterranean basin live about 210 elasmobranch species including sharks, skates and rays. Certain species are present in both the Northeast Atlantic and the Mediterranean basin. In Table 5.1 a summary of the number species for each group and area is presented.

Table 5.1. Number of elasmobranch species described for the NE Atlantic and Mediterranean.

Group NE Atlantic Mediterranean Total

SHARKS 78 (+ 2?) 46 (+ 3?) 124 (+ 5?)

SKATES and RAYS 49 36 (+ 2?) 85 (+ 2?)

Total 127 (+ 2?) 82 (+ 5?) 209 (+ 7?)

The MEDITS program, which is included in the European community Data Collec-tion Framework, it is working on the production of an atlas of the maturity scales, which includes images of all the elasmobranches living in the Mediterranean and Black Sea. In a short time such atlas will be published and shared worldwide by in-ternet. Based on such experience, it is considered useful and necessary the production of a similar tool, for the ICES areas, that will allow sharing information and know-how among all the scientists involved in field scientific studies of fishing resources in European waters.

As a first step, it is needed to build a reliable faunistic list that represents the species living in both considered areas. For such objective, it was decided to take under con-sideration the elasmobranch faunistic list of the NE Atlantic and the Mediterranean that refers to the Eschmeier catalogue.

With the final objective of compiling that Atlas to illustrate the maturity scales, WKMSEL 2 participants provided photos for a wide number of species. For this com-pilation it was taken into account the variability of reproductive strategies that exists among elasmobranch species. Two main groups were considered, but taken into ac-count the differences between each one of the reproductive strategies within each group:

- oviparity, which includes single and multiple oviparity (both lecithotrophic);

- viviparity, which includes one type of lecithotrophy, yolk-sac viviparity, and six types of matrotrophy: oophagy, placental viviparity, lipid histotrophy, limited histotrophy, lamnid oophagy and carcharinid oophagy (Hamlett, 2005).

During the meeting, a subgroup was assigned to make the selection of the best photos to illustrate the scales proposed by WKMSEL 2. Starting from the compilation from the first WKMSEL meeting, the new photos were incorporated to fill the missing gaps, and increasing the number of species and reproductive strategies covered (see Annex 3).

A table with the list of species present in Northeast Atlantic and the Mediterranean associated to the availability of the photos for each maturity stage was compiled and


made available on the sharepoint (“Maturity stages_summary of available photos.xls”). This same file will be made available along with this report on the WKMSEL2 website (http://ices.dk/workinggroups/ViewWorkingGroup.aspx?ID=587). Moreover a tem-plate sheet was developed by participants to standardize the visualization of the pho-tos for each maturity stage scale by species. In particular it was possible to create a photo-template for the following reproductive strategies and key species:

Lecithotrophic - single oviparity: S. canicula; Raja brachyura; Raja clavata;

Lecithotrophic - multiple oviparity: G. melastomus;

Lecithotrophic - yolk-sac viviparity: E. spinax; S. rostratus;

Matrotrophic – viviparity (different reproductive strategies and species in-cluded)

These templates were divided by sex in order to be as useful as possible for a macro-scopic determination of maturity stages during fieldwork, onboard or in laboratory. For the remaining species, for which photos are not available to illustrate all stages, the selection made by the group could incorporate future templates when a complete collection is attained. In particular it is recommended to improve the collection of photos on matrotrophic viviparity. The photo collection is presented in Annexes 5 and 6.

6 Guidelines for an optimal sampling strategy for the determina-tion of maturity stages in elasmobranchs

In response to the ToR “propose an optimal sampling strategy to estimate accurately the maturity ogives”, two ICES documents were consulted: the guidelines for collect-ing maturity data and histological analyses for maturity workshops developed by the Planning Group on Commercial Catch, Discards and Biological Sampling (PGCCDBS) meeting in 2010 and the recommendations from the report of the Work-shop on Maturity Ogive Estimation for Stock Assessment (ICES WKMOG 2008). From the experience gathered by the participating countries the following guidelines to collect data for maturity estimation of elasmobranch fishes were developed:

1 ) Sampling for many species/stocks may need to be conducted in coopera-tion among the participating laboratories, in order to collate sufficient data.

2 ) The number of samples by length range, sex, location and season has to be clearly defined, considering the number of countries involved, timing, and spatial overlap of sampling.

3 ) Preferably, the sampling procedures should be conducted several times during a year (preferably covering all the seasons) in order to assess the reproductive cycle and development of the gonads. Hence, sampling should be conducted at least 4 times at year, or more frequently depending on species, reproductive mode and area.

4 ) Given that fishery-independent trawl surveys are not usually conducted each quarter, limitations in sampling capacity are recognised. Commercial fleet sampling (e.g., from onboard observer schemes) could be alternative-ly used to complement the existing samples if observers are properly trained for maturity staging. For this, having clear maturity stages defini-tions, based on macroscopic observations and supported by imag-es/photographs that can be used by fishery observers onboard commercial vessels, is essential.

http://ices.dk/workinggroups/ViewWorkingGroup.aspx?ID=587


5 ) Improve the training for maturity staging, with frequent calibration work-shops conducted within and across laboratories, and collection of quantita-tive measure of maturity to reduce bias on stage assignment.

6 ) For data collection, each specimen should be given a fish ID including the following information: country, trawl station, date and fish number etc., as suggested by PGCCDBS 2010.

7 ) Staging assignment should be performed on fresh samples, when possible. Staging from pictures or preserved/frozen specimens is more difficult and may lead to errors, hence this should be avoided.

8 ) For survey data to be used in a maturity index of the “spawning stock” (term used to follow adopted terminology for stock assessment), the sur-vey must be conducted at the right time compared to the peak of the re-productive period and have adequate coverage. If survey data are not available at the right time then validated maturity data obtained outside the peak of the reproductive season can be used, although this should be confirmed on a stock-by-stock basis.

9 ) Where valid maturity data are available from market samples it can be used to estimate maturity. This is mainly the case for species with a pro-tracted reproductive season where survey data do not cover the whole sea-son or stock area. Also, if survey and market data do not show systematic differences it can be used together. However, for the species gutted before landing, market sampling may only allow the collection of maturity infor-mation for males, and only based on clasper state.

10 ) Maturity data from market samples should be collected during the whole pre-reproductive season (for determinate species) or peak of the reproduc-tive season (for indeterminate species) season on a métier based sampling programme, and cover the whole stock distribution area.

11 ) As with market samples, onboard samples should be collected on a métier-base to avoid gear and fleet selectivity effects and in the correct time and spatial frame compared to the time of reproduction.

12 ) Whenever possible, maturity staging should take place onboard the survey vessel.

13 ) A comprehensive illustrated guide should be available for all stocks re-quiring maturity observations.

14 ) Macroscopic maturity scales used should be validated through the collec-tion of quantitative measure of maturity, histologically or in another ap-propriate way.

15 ) Data collected should be plotted and mapped to assess differences by source, strata, location and time.

16 ) Length stratified maturity data should be weighted by the length distribu-tion. If samples are collected on a random scheme or the stock is assessed on a length basis, no weighting according to the length distribution is re-quired.

17 ) If the fish maturation process is dependent on age and/or sex as well as length then a Sex Maturity Age Length Key (SMALK) should be used. Age reading precision is important in this context.

18 ) If the stock shows a sexual difference in maturity a female maturity ogive should be used, or the effect of combining both sexes considered in detail.


19 ) Estimation of the maturity ogive should take into account precision and bias in both the sampling and estimation process as described in the ICES WKMOG REPORT 2008.

7 Applications and other data collection

7.1 Introduction

The group tried to identify the information on reproductive biology (e.g. maturity stages) that are most valuable for fisheries management and stock assessment, and such information includes the basic life-history parameters needed in demographic and other assessment models, and spatial information, that may aid in informing al-ternative management actions. This table should be considered a first draft, as it needs to be circulated among experts for further comment before being finalized.

7.2 Demographic information

The main types of reproductive information of most relevance for stock assessment are summarized in Table 7.1.

Furthermore, in addition to spatial information on life-history stages (see Section 7.3), providing that sampling protocols are standardised across study areas (in terms of methods and time of sampling), then spatial differences in life history parameters can also be used to inform on stock identity. Temporal differences in demographic pa-rameters are discussed briefly in Section 7.4.

Table 7.1: Reproductive parameters for elasmobranchs

Information OVI VIV Notes

1

Age/Length-at-maturity

X X

All the maturity stages described above can be categorized as Immature (stages 1-2) and biologically Mature (Stages 3, 4, 5 and 6). This information is needed to provide the length (age) of the smallest mature fish, when 50% are mature (L50%) and the length of the largest immature fish.

2

Age/Length-at-maternity

X X

Maternity ogive is the relationship between the proportion of the female population contributing to annual recruitment and the age/size of the animals. This is the most complex component to determine, as it inevitably requires information on periodicity of each of the ovarian cycle and gestation before estimating values of the parameters of the ogive (Walker, 2005). So, according to the definition we cannot state what stages to use without prior information about the reproductive cycle since it is extremely dependent on the periodicity of the cycle. In oviparous species it can use information from stages 3b to 4a, where for viviparous from stages 4 to 6. Depending on the periodicity of the reproductive cycle. It would be expected that the length of the smallest fish actively reproducing would be slightly greater than the length at first maturity. The proportion of the stock that is actively reproducing will also be affected by reproductive mode and cycle, including resting years.

3 Fecundity (including how fecundity is

X

Captive observations: Maintaining captive fish in aquarium facilities can allow the egg-laying rates and annual egg production to be calculated over the spawning season, and has been used for several rajids, especially when the egg-


correlated with length and/or age)

laying season is a few months (see Ellis and Shackley, 1995; Chevolot et al., 2007; Palm et al., 2011). This can provide fecundity at length information, although it is unknown if conditions in captivity may affect egg-laying.

X X

Ovarian fecundity: Mature follicles can be counted to estimate the number of young that may be produced. Sampling should ideally be undertaken monthly, so that the reproductive cycle and atresia can be monitored. Females on stages 3a and 3b in oviparous and 3 (or 3-4 in viviparous when vitellogenesis is synchronous with gestation) in viviparous should be considered. This method is widely used to estimate fecundity at length, although it must be recognized that not all mature follicles will successfully be fertilized and develop. Such studies should also consider the reproductive cycle and whether or not vitellogenesis is synchronous with gestation. In oophagic species ovarian fecundity should not be used.

X

Uterine fecundity: The numbers of embryos/pups in early, mid and late-pregnancy (Stages 4a,b,c). Several studies have indicated that uterine fecundity is less than ovarian fecundity. This may be due to ovarian fecundity over-estimating fecundity (e.g. not all mature follicles are ovulated), but it is also possible for some pups to be ‘aborted’ during capture (i.e. uterine fecundity may under-estimate fecundity).

4

Reproductive cycle X X

Although the reproductive cycle of males is generally considered to be annual, females may have reproductive cycles of 1-3 years, depending on reproductive mode, the gestation period, and whether or not vitellogenesis and gestation are synchronous. The reproductive cycle can be examined with various measures, such as gonad state (e.g. gonadosomatic index, size distribution of follicles, levels of atresia), uterine state (e.g. maturity stage, size distribution of pups), as well as through endocrinology (e.g. levels of reproductive hormones).

5

Size-at-hatching X

The length at which the young emerge from the egg-case. This can be determined through maintaining egg-cases in aquaria, and also from any observations of wild fish that have hatched recently (e.g. they may still contain a small external or internal yolk-sac).

6

Size-at-birth X

The length at which the young are born. This can be determined through examination of females at stage 4c (later-term), that present fully developed, and also from any observations of wild fish that have recently been born (i.e. those specimens that still have a fresh umbilical scar).

7

Sex-ratio of litters X X

Although the sex-ratio is generally not different to the expected 1:1 ratio, any dedicated studies should report this. For oviparous species, this can only be determined from aquarium observations. For viviparous species, the sex-ratio can be confirmed from examination of embryos in the uteri (i.e. from females as stages 4b,c).


7.3 Spatial information

Spatial management of important elasmobranch habitats has been suggested as a po-tentially useful method for the improved management of their stocks (Bonfil, 1999; Hunter et al., 2006). Although our knowledge of critical elasmobranch habitats is lim-ited for many species, important habitats for elasmobranch fish may include sites/depths of importance for reproduction and feeding (e.g. Coelho and Erzini, 2010), as well as migration corridors. Mating grounds (Carrier and Pratt, 1998; Cam-pana et al., 2003), pupping areas (McCandless et al., 2002), spawning beds for ovipa-rous species (Ellis and Shackley, 1997; Ellis et al., 2005) and nursery grounds (Castro, 1993; Simpfendorfer and Milward, 1993; Ellis et al., 2005) may all be viewed as ecolog-ically important habitats for elasmobranch stocks.

The egg-laying (or oviposition) sites of oviparous elasmobranchs may be important habitats, especially if such sites require particular habitat characteristics, are limited in spatial extent, and are used consistently between years. There is anecdotal evi-dence suggesting that rajids and other oviparous chondrichthyans, including chimae-riforms, heterodontiform sharks and some orectolobiform and carcharhiniform sharks, have discrete spawning beds (Dean, 1906; Williamson, 1913; Orton, 1926; Smith, 1942; Hitz, 1964; Able and Flescher, 1991; Chembian, 2007; Flammang et al., 2011). For example, McLaughlin and O’Gower (1971) reported that the eggs of the Port Jackson shark Heterodontus portusjacksoni were deposited in traditional oviposi-tion sites on shallow, sheltered reefs, and Able and Flescher (1991) reported 300 egg cases of the chain catshark Scyliorhinus retifer attached to the hydroid Eudendrium, suggesting that eggs were laid in structured habitats. Lesser-spotted dogfish S. ca-nicula also deposit their eggs on a variety of upright structures, including macroalgae and, on offshore grounds, erect sponges, hydroids, soft corals and bryozoans (Ellis and Shackley, 1997).

In terms of skates, although the egg-laying season can be relatively protracted (Hold-en, 1975) and spawning migrations have been suggested to occur (e.g. Holden 1975; Hunter et al., 2005a,b), little is known about the sites of spawning grounds (Ellis et al., 2005, 2012), although Holden (1975) noted that parts of the Wash were grounds where female thornback ray Raja clavata would congregate. Rajid egg cases have horns and an “adhesive film” for anchorage, but little is known about the types of substrates on which they are laid and whether certain sites are preferred. Anecdotal-ly, Williamson (1913) reported that large numbers of skate eggs were taken off the shoal water on Aberdeen Bank. Similarly, large numbers of egg cases (up to 152 per 30 minute tow of a scallop dredge) of ‘Raja’ binoculata were reported off the coast of Oregon (Hitz, 1964), and potential spawning grounds have been noted for other spe-cies along the western coasts of North America (Hoff, 2008, 2010; Love et al., 2008) and elsewhere in the Pacific (Hunt et al., 2011).

Improved identification guides for egg cases of oviparous species are needed to facili-tate accurate identification of viable egg cases that may be taken in trawl surveys.

7.4 Temporal information

Several studies have examined potential temporal differences in maturity and fecun-dity in elasmobranchs (e.g. Ellis and Keable, 2008; McCully et al., 2012, and references therein), as such information can inform on potential population responses to over-exploitation. In most cases, determining whether there has been a genuine change in, for example, the length at maturity, has been hampered by different studies using


different maturity scales, highlighting the need for a standardised approach to col-lecting reproductive parameters.

8 Conclusion and recommendations

The maturity scales developed by WKMSEL 2010 were updated and revised for male and female oviparous and viviparous species. Modifications were based on published literature, and the knowledge and experience of the participants attending the work-shop.

The revision of the scales was made taking into account a greater number of species and reproductive strategies than in WKMSEL 2010.

A comparison and conversion between the existing maturity scales (including WKM-SEL 2010) and the WKMSEL 2012 ones was undertaken in order to provide a com-mon tool for exchanging data and scientific information, as well as to help on database updates.

In general, changes carried out were related to:

1 ) Terminology. The group agreed to adopt the terminology proposed by Brown-Peterson et al. 2011. However, terms such as “spawning” and “re-gressing” were considered inadequate for the description of elasmobranch reproduction in particular, and may therefore not be accepted by the scien-tific community. Instead, the term “active“ was adopted for males, “egg-laying” and “post-laying” were adopted for oviparous and “post-partum” for viviparous (see section 3.1 for more details).

2 ) Description of structures. The description of each stage was rewrote, in order to make it simpler and ready-to-use: 2.1 ) The name of each specific structure was highlighted in the scale (in

bold, and in separate lines); 2.2 ) Every detail that was not essential for assessing the stage was re-

moved; 2.3 ) We avoided using subjective descriptive terms, like the use of col-

ours to describe stages (e.g. “yellow” for ovarian follicles and “red-dish” for claspers).

3 ) Transition among stages. The transition between stages was clarified taking into account the application of maturity data for data analysis used for stock assessment. 3.1 ) Regarding the oviparous females scale. According to the experience

gathered since the last workshop, the participants accepted the maintenance of stages 4a and 4b, and their description was updated.

3.2 ) Regarding the viviparous females scale: The difference between ma-ture and maternal conditions was clarified. This is essential, in par-ticular, for determining the periodicity of the reproductive cycle. The numeric system of the stages was amended accordingly (see chapter 3.2 for more details).

Participants recommended to:

4 ) Continue to collect information on more species especially those that attain relatively large size, such as pelagic elasmobranchs;


5 ) Collect more information on all the different viviparous modes of repro-duction, particularly lamnoid and carcharhinid oophagy and histotrophy (both limited and lipid);

6 ) From now onward, analyse maturity stage data according to the scales herein proposed in order to validate its application for stock assessment modelling (see chapter 7 for more details);

7 ) Increase the geographical range of the data collection, including infor-mation from the Atlantic, North Sea, Baltic and Eastern and Southern Med-iterranean countries. Information from long distance fisheries (e.g. Pacific, Arctic etc.) exploited by European fleets would also be welcome;

8 ) Promote a proper dissemination of the proposed scale to all ICES and Mediterranean countries, but also to other geographical areas, through one or more of the following hypothesis: 8.1 ) Journal article (proposal: ICES Journal of Marine Science) 8.2 ) Publish the scale and Atlas through ICES website (with possible ex-

change of photos with other scientists) – the ICES will be contact by email about this option.

8.3 ) FAO Technical Paper 8.4 ) The option to upload the Atlas photos on WebGR should be ex-

plored. If someone already used or know anyone that used this tool can you please give us your feedback (http://webgr.wiki.azti.es/doku.php) (see Annex 7).

9 ) Promote calibration workshops inside and between laboratories.

As a final and important recommendation, it is proposed that the new maturity scales be revised and commented by the ICES Working groups responsible for data collec-tion, like PGCCDBS, IBTSWG and WGBEAM, as well as by the advisory groups for elasmobranchs WGEF, before further dissemination inside and outside the ICES.

http://webgr.wiki.azti.es/doku.php


9 References and working documents

Aasen, O. 1966. Bllihaien. Prionace glauca (Linnaeus) 1758. Fisken Havet 1:1·16.

Able, K. W. and Flescher, D. 1991. Distribution and habitat of chain dogfish, Scyliorhinus retifer, in the mid-Atlantic Bight. Copeia, 1991(1): 231–234.

Bonfil, R. 1999. Marine protected areas as a shark fisheries management tool. Proceedings of the 5th Indo-Pacific Fish Conference, Nouméa, 1997 (B. Séret and J.-Y. Sire, eds), 217–230.

Brown-Peterson, N.J., Lowerre-Barbieri, S.K., Macewicz, B.J., Saborido-Rey, F., Tomkiewicz, J. and Wyanski, D.M. 2011. A Standardized Terminology for Describing Reproductive De-velopment in Fishes. Marine and Coastal Fisheries: Dynamics, Management, and Ecosys-tem Science 3: 52-70.

Campana, S., Joyce, W. and Marks, L. 2003. Status of the porbeagle shark (Lamna nasus) popula-tion in the Northwest Atlantic in the context of species at risk. Canadian Science Advisory Secretariat, Research Document 2003/007, 27 pp.

Carrier, J. C. and Pratt, H. L. 1998. Habitat management and closure of a nurse shark breeding and nursery ground. Fisheries Research 39, 209–213.

Castro, J. I. 1993. The shark nursery of Bulls Bay, South Carolina, with a review of the shark nurseries of the southeastern coast of the United States. Environmental Biology of Fishes, 38: 37–48.

Chembian, A. J. 2007. New record of Rhinochimaera atlantica (Chimaeriformes: Rhinochimaeri-dae) spawning ground in the Gulf of Mannar along the south-east coast of India. Indian Journal of Fisheries, 54: 345–350.

Chevolot, M., Ellis J.R., Rijnsdorp, A.D., Stam W.T. and Olsen J.L. 2007. Multiple paternity analysis in the thornback ray Raja clavata L. Journal of Heredity, 98: 712–715.

Coelho, R. and Erzini, K. 2006. Reproductive aspects of the undulate ray, Raja undulata, from the south coast of Portugal Fisheries Research, 81: 80-85

Dean, B. 1906. Chimaeroid fishes and their development. Carnegie Institution of Washington, 194pp.

Ellis, J. R., Cruz-Martinez, A., Rackham, B. D. and Rogers, S. I. 2005. The distribution of chon-drichthyan fishes around the British Isles and implications for conservation. Journal of Northwest Atlantic Fishery Science, 35: 195–213.

Ellis, J. R. and Keable, J. 2008. The fecundity of Northeast Atlantic spurdog (Squalus acanthias L., 1758). ICES Journal of Marine Science, 65: 979–981.

Ellis, J. R., Milligan, S. P., Readdy, L., Taylor N. and Brown M. J. 2012. Spawning and nursery grounds of selected fish species in UK waters. Science Series Technical Report, CEFAS Lowestoft, 147, 56 pp.

Ellis, J. R. and Shackley, S. E. 1995. Observations on egg-laying in the thornback ray. Journal of Fish Biology, 46: 903-904.

Ellis, J. R. and Shackley, S. E. 1997. The reproductive biology of Scyliorhinus canicula in the Bris-tol Channel, U.K. Journal of Fish Biology, 51: 361–372.

Figueiredo, I., Moura, T., Neves, A. and Gordo, L.S. 2008. Reproductive strategy of leafscale gulper shark, Centrophorus squamosus, and Portuguese Dogfish, Centroscymnus coelolepis, on the Portuguese continental slope. Journal of Fish Biology, 73: 206-225.

Flammang, B. E., Ebert, D. A. and Cailliet, G. M. 2011. Intraspecific and interspecific spatial distribution of three eastern North Pacific catshark species and their egg cases (Chondrich-thyes: Scyliorhinidae). Breviora, 525: 1–18.


Hamlett, W.C. 2005. Reproductive biology and phylogeny of Chondrichthyes: sharks, batoids and chimaeras. Sciences Publishers, Inc, New Hampshire, USA.

Hitz, C. R. 1964. Observations on the egg cases of the big skate (Raja binoculata Girard) found in Oregon coastal waters. Journal of the Fisheries Research Board of Canada, 21: 851–854.

Hoff, G. R. 2008. A nursery site of the Alaska skate (Bathyraja parmifera) in the eastern Bering Sea. Fishery Bulletin, 106: 233–244.

Hoff, G. R. 2010. Identification of skate nursery habitat in the eastern Bering Sea. Marine Ecolo-gy Progress Series, 403: 243–254.

Holden, M. J. 1975. The fecundity of Raja clavata in British waters. Journal du Conseil, 36: 110–118.

Holden M.J. and Raitt D.F.S. 1974. Manual of fisheries sciences, Pt 2: Methods of resource in-vestigation and their application. FAO Fisheries Technical paper No 115, revision 1. Unit-ed Nations Development Programme, food and Agriculture Organization of the United Nations, Italy. pp 127-133.

Hunt, J. C., Lindsay, D. J. and Shahalemi, R. R. 2011. A nursery site of the golden skate (Raj-iformes: Rajidae: Bathyraja smirnovi) on the Shiribeshi Seamount, Sea of Japan. Marine Bio-diversity Records, 4, e70, 7pp. (Published online)

Hunter, E., Berry, F., Buckley, A. A, Stewart, C. and Metcalfe, J. D. 2006. Seasonal migration of thornback rays and implications for closure management. Journal of Applied Ecology, 43: 710–720.

Hunter, E., Buckley, A. A., Stewart, C. and Metcalfe, J. D. 2005a. Migratory behaviour of the thornback ray, Raja clavata, in the southern North Sea. Journal of the Marine Biological As-sociation of the United Kingdom, 85: 1095–1105.

Hunter, E., Buckley, A. A., Stewart, C. and Metcalfe, J. D. 2005b. Repeated seasonal migration by a thornback ray in the southern North Sea. Journal of the Marine Biological Association of the United Kingdom, 85: 1999–1200.

ICES. 2008. Report of the Workshop on Maturity Ogive Estimation for Stock Assessment (WKMOG), 3-6 June 2008, Lisbon, Portugal. ICES CM2008/ACOM:33. 72 pp.

ICES. 2009. Report of the Joint Meeting between ICES Working Group on Elasmobranch Fishes (WGEF) and ICCAT Shark Subgroup. ICES CM 2009/ACOM:16. Copenhagen, Denmark. 405 pp.

ICES. 2010. Report of the Workshop on Maturity Staging for Elasmobranchs (WKMSEL). ICES CM 2010/ACOM 48. 132 pp.

Love, M. S., Schroeder, D. M., Snook, L., York, A. and Cochrane, G. 2008. All their eggs in one basket: a rocky reef nursery for the longnose skate (Raja rhina Jordan and Gilbert, 1880) in the southern California Bight. Fishery Bulletin, 106: 471–475.

McCandless, C.T., Pratt, H.L. Jr. and Kohler, N.E. 2002. Shark nursery grounds of the Gulf of Mexico and the East Coast waters of the United States: an overview. An internal report to NOAA’s Highly Migratory Species Office. NOAA Fisheries Narragansett Laboratory, 28 Tarzwell Dr., Narragansett, RI 02882. 287 pp.

McCully, S. R., Scott F. and Ellis J. R. 2012. Length at maturity and conversion factors for skates (Rajidae) around the British Isles, with a critique of earlier studies. ICES Journal of Marine Science, 69: 1812–1822.

Moura, T., Nunes, C., Bandarra, N., Gordo, L. S., Figueiredo, I. (2011). Embryonic development and maternal-embryo relationships of the Portuguese dogfish Centroscymnus coelolepis. Marine Biology 158:401–412.

Orton, J. H. 1926. A breeding ground of the nursehound (Scyliorhinus stellaris) in the Fal Estu-ary. Nature, 118: 732.


Palm, B. D., Koester, D. M., Driggers, W. B. and Sulikowski, J. A. 2011. Seasonal variation in fecundity, egg case viability, gestation, and neonate size for little skates, Leucoraja erina-cea, in the Gulf of Maine. Environmental Biology of Fishes, 92: 585–589.

Pratt, H. L.J. 1988. Elasmobranch gonad structure: a description and survey. Copeia, (3): 719-729.

Relini G., Bertrand J. and Zamboni A. 1999. Synthesis of the knowledge on bottom fishery re-sources in central Mediterranean (Italy and Corsica). Biol. Mar. Medit. 6(suppl. 1): 868pp.

Relini G., Carpentieri P. and Murenu M. 2008. Manuale di Istruzioni Medits (Medits Instruction Manual). Biologia Marina Mediterranea, 15 (Suppl. 2): 1-78.

Serra-Pereira B., Figueiredo I. and Gordo L.S. 2011. Maturation of the Gonads and Reproduc-tive Tracts of the Thornback Ray Raja clavata, with Comments on the Development of a Standardized Reproductive Terminology for Oviparous Elasmobranchs. Marine and Coastal Fisheries: Dynamics, Management, and Ecosystem Science 3: 160-175.

Simpfendorfer, C. A. and Milward, N. E. 1993. Utilisation of a tropical bay as a nursery area by sharks of the families Carcharhinidae and Sphynidae. Environmental Biology of Fishes, 37: 337–345.

Smith, B. G. 1942. The heterodontid sharks: Their natural history, and the external develop-ment of Heterodontus japonicus based on notes and drawings by Bashford Dean. The Bash-ford Dean Memorial Volume Archaic Fishes, Article VIII. American Museum of Natural History, New York, 647–784.

Stehmann, M. 1987. Quick and dirty tabulation of stomach contents and maturity stages for skates (Rajidae), squaloid and other ovoviviparous and viviparous species of sharks. Pages 5–9 in American Elasmobranch Society Newsletter 3.

Stehmann, M.F.W. 2002. Proposal of a maturity stages scale for oviparous and viviparous carti-laginous fishes (Pisces, Chondrichthyes). Archive of Fishery and Marine Research, 50 (1): 23-48.

Ungaro N. 2008. Field manual on macroscopic identification of maturity stages for the Mediter-ranean fishery resources. GCP/RER/ITA/MSM-TD-21. MedSudMed Technical Documents No 21: 34 pp

Williamson, H. C. 1913. On the eggs of certain skates (Raia). Scientific Investigations, Fishery Board for Scotland, Part I, 3–6.


Annex 1: List of participants

Country Name Address Phone/Fax Email GRC Aikaterini

Anastasopoulou Hellenic Centre For Marine Research (HCMR) 46.7 Km Athens Sounio, Mavro Lithari, PO BOX 712 19013 Anavissos, Attica Greece

Phone +30 2109856705 Fax +30 2109811713

[email protected]

BRA André Afonso Universidade do Algarve / Universidade Federal Rural de Pernambuco (UFRPE) Av. Dom Manuel de Mederios, s/n 52171-900 Recife, Pernambuco Brazil

Phone +55 8181062323 Phone +351964288393

[email protected]

PRT Barbara Serra Pereira (co-chair)

Instituto Português do Mar e da Atmosfera (IPMA) Avenida de Brasilia 1449-006 Lisbon Portugal

Phone +351 21 3027108

[email protected]

PRT Catarina Maia


Phone +351 21 3027108

[email protected]

ITA Cecilia Mancusi (corresponding participant)

ARPAT - Tuscany Regional Agency for Environmental Protection via Marradi 114 57126 Livorno - Italy

Phone +39 339 3462217

[email protected]

AUS Fabian Trinnie Fisheries Research Branch, Fisheries Victoria 2A Bellarine Hwy Queenscliff, 3225 Victoria Australia

Phone +61 0352580255 Fax +61 0402518322

[email protected]

ITA Fabrizio Serena (co-chair)


Phone +39 3204391149

[email protected]

DZA Farid Hemida ENSSMAL (Ecole Nationale Supérieure des Sciences de la Mer et de l'Aménagement du littoral) Campus universitaire - BP 19 Bois des Cars- Dely Ibrahim 16320 Algiers Algeria

Phone +213 771577999

[email protected]


ESP Gabriel Morey (corresponding participant)

Fisheries Department, Balearic Islands Government c/Foners, 10 07006 Palma de Mallorca Spain

[email protected]

GBR Jim Ellis (corresponding participant)

CEFAS, Lowestoft Laboratory Lowestoft NR33 0HT Suffolk United Kingdom

[email protected]

PRT Joana Fernandez-Carvalho

Instituto Português do Mar e da Atmosfera (IPMA) Av 5 de Outubro, s/n 8700-305 Olhão Portugal

[email protected]

ITA Maria Cristina Follesa

Univeristy of Cagliari - Department of Life science and Environment Via T.Fiorelli, 1 09131 Cagliari ITALY

[email protected]

BRA Maria Cristina Oddone

Instituto de Ciências Biologicas, Universidade Federal do Rio Grande (FURG) Avenida Itália, Km 8, Campus Carreiros 96203-900 Rio Grande, RS Brasil

Phone + 55 5332336633

[email protected]

BRA Mariana Rego Universidade Federal Rural de Pernambuco (UFRPE) Av. Dom Manuel de Mederios, s/n 52171-900 Recife, Pernambuco Brazil

[email protected]

ITA Massimiliano Bottaro (corresponding participant)

Italian National Institute for Environmental Protection and Research ISPRA (formerly ICRAM) Via Vitaliano Brancati, 60 I-00144 Rome Italy

Phone +39 06 50074765

[email protected]

ITA Monica Barone


Phone +39 3332240666

[email protected]

PRT Neide Lagarto Instituto Português do Mar e da Atmosfera (IPMA) Avenida de Brasilia 1449-006 Lisbon Portugal

Phone +351 21 3027116

[email protected]

SLO Polona Pengal Fisheries Research Institute of Slovenia Spodnje Gameljne 61a 1000 Ljubljana Slovenia

[email protected]

mailto:[email protected]

mailto:[email protected]


ITA Rita Cannas Univeristy of Cagliari - Department of Life science and Environment Via T.Fiorelli, 1 09131 Cagliari ITALY

[email protected]

PRT Rui Coelho Instituto Português do Mar e da Atmosfera (IPMA) Av 5 de Outubro, s/n 8700-305 Olhão Portugal

[email protected]

PRT Sérgio Amorim Instituto Português do Mar e da Atmosfera (IPMA) Av. General Norton de Matos Nº4 4450-208 Matosinhos

Phone +351 936861127

[email protected]

GBR Sophy McCully (corresponding participant)

CEFAS, Lowestoft Laboratory Lowestoft NR33 0HT Suffolk United Kingdom

[email protected]

PRT Teresa Moura


Phone +351 21 3027131

[email protected]

SLO Tomaz Modic

Fisheries Research Institute of Slovenia Spodnje Gameljne 61a 1000 Ljubljana Slovenia

[email protected]


Annex 2: Agenda

Tuesday, 11th December 2012

9.00 – 10.30 Presentation of chairs and participants.

Adoption of the agenda and time-table.

Overview of the previous WKMSEL achievements

10.30 -11.00 Coffee break

11.00 - 13.00 Presentation by participant:

i) the use of proposed maturity scales (pros and cons);

ii) other maturity scales still in use;

iii) adopted protocol for data collection;

iv) application of maturity scales data.

(5-10 min each)

13.00 – 14.00 Lunch break

14.00 – 15.30 Group discussion about the adopted maturity scales for oviparous and viviparous elasmobranchs

15.30 – 16.00 Coffee break

16.00 – 17.30 Continue from previous session

Workplan by subgroup:

- Oviparous maturity scale; - Viviparous maturity scale; - Atlas to illustrate the use of the adopted maturity scales by

species or group of species;

Wednesday, 12th December

9.00 – 10.30 Subgroup sessions:

- Define the list of species (sharks, skates and rays) and types of reproduction

- Revise the maturity scales proposed by WKMSEL - Compile photo Atlas


11.00 - 13.00 Continue with sub-group sessions


14.00 – 15.30 Plenary



16.00 – 17.30 Continue with sub-group sessions

Thursday, 13th December

9.00 – 10.30 Continue with sub-group sessions


11.00 - 13.00 Continue with sub-group sessions


14.00 – 15.30 Plenary


16.00 – 17.30 Presentation of the sub-groups conclusions

Friday, 14th December

9.00 – 10.30 Continue from the previous session.


11.00 - 13.00 Report


14.00 – 15.30 Report


16.00 – 17.30 List of recommendations and terms of reference for the next work-shop


Annex 3: Species list with info on the study areas and mode of reproduction

GSA Area 4 Algeria 5 Balearic Island 9 North Tyrrhenian Sea 11 Sardinia 14 Gulf of Gabes 15 Malta Islands 16 South of Sicily 17 North Adriatic 19 Ionian Sea 22 Aegean Sea ICES Area IXa Portugal FAO Area 34 41


OVIPAROUS SPECIES Areas (GSA, ICES or FAO)

Group Order Family Species Macro Micro Mode of reproduction (Hamlet, 2005) Sharks Carcharhiniformes Scyliorhinidae Scyliorhinus canicula GSA 9, 19, 22 GSA 9 Lecithotrophic - single oviparity

Sharks Carcharhiniformes Scyliorhinidae Scyliorhinus stellaris GSA 5, 15,9 GSA 5 Lecithotrophic - single oviparity

Skates Carcharhiniformes Scyliorhinidae Galeus melastomus GSA 5,9,11,19, 22, ICESIXa

GSA 11 Lecithotrophic - multiple oviparity

Skates Carcharhiniformes Scyliorhinidae Galeus atlanticus ICESIXa Lecithotrophic - single oviparity

Skates Rajiformes Arhynchobatidae Rioraja agassizi FAO 41 Lecithotrophic - single oviparity

Skates Rajiformes Rajidae Atlantoraja castelnaui FAO 41 Lecithotrophic - single oviparity

Skates Rajiformes Rajidae Atlantoraja cyclohpora FAO 41 Lecithotrophic - single oviparity

Skates Rajiformes Rajidae Atlantoraja platana FAO 41 Lecithotrophic - single oviparity

Skates Rajiformes Rajidae Dipturus oxyrinchus GSA 9, 4 Lecithotrophic - single oviparity

Skates Rajiformes Rajidae Dipturus nidarosiensis GSA 4 Lecithotrophic - single oviparity

Skates Rajiformes Rajidae Leucoraja circularis GSA 5, 9 Lecithotrophic - single oviparity

Skates Rajiformes Rajidae Leucoraja naevus GSA 5; ICESIXa Lecithotrophic - single oviparity

Skates Rajiformes Rajidae Raja asterias GSA 4,9 GSA 9 Lecithotrophic - single oviparity

Skates Rajiformes Rajidae Raja brachyura GSA 5, 9; ICESIXa Lecithotrophic - single oviparity

Skates Rajiformes Rajidae Raja clavata GSA 5, 9, 15;ICES IXa GSA 9; ICESIXa

Lecithotrophic - single oviparity

Skates Rajiformes Rajidae Raja miraletus GSA 5, 9, 15, 19 GSA 9 Lecithotrophic - single oviparity

Skates Rajiformes Rajidae Raja microcellata ICESIXa Lecithotrophic

Skates Rajiformes Rajidae Raja montagui ICES IXa Lecithotrophic - single oviparity

Skates Rajiformes Rajidae Raja polystigma GSA 5, 9 Lecithotrophic - single oviparity

Skates Rajiformes Rajidae Raja radula GSA 5 Lecithotrophic - single oviparity

Skates Rajiformes Rajidae Raja undulata ICES IXa Lecithotrophic - single oviparity

Skates Rajiformes Rajidae Rostroraja alba GSA 5, 9 GSA 5 Lecithotrophic - single oviparity

Skates Rajiformes Rajidae Sympterygia acuta FAO 41 Lecithotrophic - single oviparity


VIVIPAROUS SPECIES areas

Group Order Family Species Macro Micro Mode of reproduction (Hamlet, 2005)

Sharks Hexanchiformes Hexanchidae Hexanchus griseus GSA 9 Lecithotrophic - yolk-sac viviparity

Sharks Hexanchiformes Hexanchidae Heptranchias perlo Lecithotrophic - yolk-sac viviparity

Sharks Squaliformes Centrophoridae Centrophorus granulosus ICES IXa; GSA 5, 9, 19 Lecithotrophic - yolk-sac viviparity

Sharks Squaliformes Centrophoridae Centrophorus squamosus ICES IXa Lecithotrophic - yolk-sac viviparity

Sharks Squaliformes Centrophoridae Deania calcea ICES IXa Presumably Matrotrophic limited Histotrophyc (?)

Sharks Squaliformes Somniosidae Centroscymnus coelolepis ICES IXa Lecithotrophic - yolk-sac viviparity

Sharks Squaliformes Somniosidae Scymnodon ringens ICES IXa Lecithotrophic - yolk-sac viviparity

Sharks Squaliformes Somniosidae Somniosus rostratus GSA 9 Lecithotrophic - yolk-sac viviparity

Sharks Squaliformes Somniosidae Centroselachus crepidater ICES IXa Lecithotrophic - yolk-sac viviparity

Sharks Squaliformes Oxynotidae Oxynotus centrina GSA 4 Lecithotrophic - yolk-sac viviparity

Sharks Squaliformes Dalatidae Dalatias licha GSA 4, 5, 9, 19 9 Lecithotrophic - yolk-sac viviparity

Sharks Squaliformes Etmopteridae Etmopterus spinax ICES Ixa, 5, 9, 19 Lecithotrophic - yolk-sac viviparity

Sharks Squaliformes Etmopteridae Etmopterus pusillus ICES IXa Lecithotrophic - yolk-sac viviparity

Sharks Squaliformes Squalidae Squalus acanthias 17 17 Lecithotrophic - yolk-sac viviparity

Sharks Squaliformes Squalidae Squalus blainville 9, 11, 15, 19, 22 11 Lecithotrophic - yolk-sac viviparity

Sharks Squatiniformes Squatinidae Squatina spp 4 Lecithotrophic - yolk-sac viviparity

Sharks Squatiniformes Squatinidae Squatina squatina 4 Lecithotrophic - yolk-sac viviparity

Sharks Orectolobiformes Ginglymostomatidae Ginglymostoma cirratum FAO 41 Lecithotrophic - yolk-sac viviparity

Sharks Lamniformes Pseudocarchariidae Pseudocarcharias kamoharai FAO 34 Matrotrophic - Lamnid oophagy

Sharks Lamniformes Alopidae Alopias vulpinus 5, 9 Matrotrophic - Lamnid oophagy

Sharks Lamniformes Cetorhinidae Cetorhinus maximus 9 Matrotrophic - Lamnid oophagy

Sharks Lamniformes Lamnidae Isurus oxyrinchus 16 16, FAO 41 Matrotrophic - Lamnid oophagy


Sharks Carcharhiniformes Carcharhinidae Carcharhinus obscurus 9 Matrotrophic - placental

Sharks Carcharhiniformes Carcharhinidae Carcharhinus plumbeus 9, 14, 16 17 Matrotrophic - placental

Sharks Carcharhiniformes Carcharhinidae Prionace glauca ICES IXa, 4, 9, 17 17 Matrotrophic - placental

Sharks Carcharhiniformes Triakidae Galeorhinus galeus 4 Lecithotrophic - yolk-sac viviparity

Sharks Carcharhiniformes Triakidae Mustelus asterias 15 Lecithotrophic - yolk-sac viviparity

Sharks Carcharhiniformes Triakidae Mustelus mustelus 4, 5, 9, 14, 15, 17 5, 17 Matrotrophic - placental

Sharks Carcharhiniformes Triakidae Mustelus punctulatus 15 Little known. Presumably matrotrophic - placental

Rays Rajformes Rhinobatidae Rhinobatos rhinobatos 14 Lecithotrophic - yolk-sac viviparity

Rays Rajformes Rhinobatidae Rhinobatos cemiculus 14 Lecithotrophic - yolk-sac viviparity

Rays Rajformes Gymnuridae Gymnura altavela 5, 14 Lecithotrophic - yolk-sac viviparity

Rays Rajformes Myliobatidae Myliobatis aquila 4, 5, 9 5 Matrotrophic - lipid histotrophy

Rays Rajformes Myliobatidae Pteromylaeus bovinus 9 Matrotrophic - lipid histotrophy

Rays Rajformes Dasyatidae Dasyatis centroura 9 Matrotrophic - lipid histotrophy

Rays Rajformes Dasyatidae Dasyatis pastinaca 5, 9 Matrotrophic - lipid histotrophy

Rays Rajformes Torpedinidae Torpedo marmorata 9, 5 Lecithotrophic - yolk-sac viviparity

Rays Rajformes Torpedinidae Torpedo nobiliana 9 Lecithotrophic - yolk-sac viviparity

Rays Rajformes Torpedinidae Torpedo torpedo 9, 19 Lecithotrophic - yolk-sac viviparity


Annex 4: Histological photo reference for oviparous and viviparous elasmobranch species

Participants presented the following updates on histological data.

VIVIPAROUS SPECIES

Microscopic analysis of the uterine villi of females of the Portuguese dogfish Cen-troscymnus coelolepis

Teresa Moura and Ivone Figueiredo (IPMA, Portugal)

The macro and microscopic analysis of the uterus of the Portuguese dogfish Cen-troscymnus coelolepis (Bocage and Capello, 1864) is available in Moura et al. (2011). This study highlights the role of the uterine villi in the reproduction of this species, particularly during the gestation period, when it greatly increases in length. Uterine villi were detected in the inner uterine wall of all the maturing and adult females but were absent in the juvenile ones. Microscopically, villi are composed by connective tissue lined by a stratified epithelium in developing, capable to reproduce and early pregnant females. With pregnancy, the epithelium of the villi becomes simple squa-mous, greatly decreasing its thickness. The number of blood vessels near the epitheli-um increases both in number and diameter. After parturition, the epithelium of the villi recovers the initial structure presented before ovulation.

Modification in the epithelium of the uterine villi of Centroscymnus coelolepis along the reproductive cycle: (a) female capable to reproduce, (b) early pregnant, (c) late preg-nant, and (d) post-partum. Staining: PAS/AB; art =arteriole, bv =blood vessel, ct =connective tissue, ep =epithelium, arrow =basal membrane. Scale bar = 50 µm.

53

Annex 5: Atlas for oviparous species

Lecithotrophic ‐ single oviparity ‐ Species: Scyliorhinus canicula FEMALE

DESCRIPTION STAGE MATURITY PHOTO

Ovaries: small and whitish. Undistinguishable ovarian follicles.

Oviducal gland: often not visible. In some species a thickening of the uteri where the gland will develop may be visible.

Uteri: thread‐like and narrow.

1 IMMATURE IMMATURE

Ovaries: follicles of different stages of development. Some small and medium sized yolked follicles may be present.

Oviducal gland: distinguishable and developing.

Uteri: enlarging.

2 DEVELOPING IMMATURE

Ovaries: presence of large yolked follicles ready to be ovulated.

Oviducal glands: fully developed.

Uteri: fully developed.


MATURE

Uteri: presence of egg capsules. 3b EGG‐LAYING MATURE

Ovaries: flaccid with few follicles of different sizes. Few large vitellogenic follicles entering atresia. POFs and atretic follicles may be visible.

Oviducal glands: fully developed but may be reduced in size.

Uteri: enlarged and flaccid.

4a POST‐LAYING MATURE not available

Ovaries: large with small and medium sized yolked follicles. Pre‐ovulatory follicles absent.


Uteri: enlarged.

4b REGENERATING MATURE not available

54

Species: Scyliorhinus canicula MALE


Claspers: flexible, non‐calcified and shorter than pelvic fins.

Testes: small and undeveloped (in skates, sometimes with visible lobules).

Ducts: straight and thread‐like.

1 IMMATURE IMMATURE

Claspers: flexible, partially calcified and usually as long as or longer than pelvic fins.*

Testes: developing (in skates, lobules clearly visible but not fully developed).

Ducts: developing and beginning to coil.


Claspers: rigid, fully calcified, and longer than pelvic fins (in some sharks they may only be as long as the pelvic fins).

Testes: fully developed (in skates, with fully formed lobules).

Ducts: tightly coiled and filled with sperm.

3a

CAPABLE

TO

REPRODUCE

MATURE

Claspers: similar to stage 3a, however with clasper glands dilated, sometimes swollen and reddish. Sperm may be present in clasper groove or glands.

Testes: similar to stage 3a.

Ducts: sperm observed inside (after a cut) or flowing out of the cloaca on pressure.

3b ACTIVE MATURE

Claspers: fully formed, similar to stage 3. Testes shrunken and flaccid, (in skates, with few visible lobules). On pressure sperm does not flow.

Sperm ducts: empty and flaccid

4a REGRESSING MATURE not available

55

Lecithotrophic ‐ single oviparity ‐ Species: Scyliorhinus canicula FEMALE

STAGE PHOTO

1

2

3a

56

3b

4a not available

4b not available

57

Species: Scyliorhinus canicula MALE

STAGE PHOTO

1

2

3a

58

Lecithotrophic ‐ single oviparity ‐ Species: Raja brachyura FEMALE





1 IMMATURE IMMATURE



Uteri: enlarging.





3a


MATURE not available





4a POST‐LAYING MATURE not available

59



Uteri: enlarged.

4b REGENERATIN

G MATURE

Species: Raja brachiura MALE





1 IMMATURE IMMATURE









MATURE

60




3b ACTIVE MATURE




61

Lecithotrophic ‐ single oviparity ‐ Species: Raja brachiura FEMALE

STAGE PHOTO

1

2

62

3a not available

3b

4a not available

63

4b

64

Species: Raja brachiura MALE

STAGE PHOTO

1

2

65

3a

3b

4 not available

6

66

Lecithotrophic ‐ single oviparity ‐ Species: Raja clavata FEMALE





1 IMMATURE IMMATURE



Uteri: enlarging.





3a


MATURE





4a POST‐LAYING MATURE

67



Uteri: enlarged.

4b REGENERATING MATURE

Species: Raja clavata MALE





1 IMMATURE IMMATURE








3a

CAPABLE

TO

REPRODUCE

MATURE




3b ACTIVE MATURE

6 8

68




69

Lecithotrophic ‐ single oviparity ‐ Species: Raja clavata FEMALE

STAGE PHOTO

1

2

70

3a

3b

4a

71

4b

72

Species: Raja clavata MALE

STAGE PHOTO

1

2

73

3a

3b

4 not available

74

Lecithotrophic ‐ multiple oviparity ‐ Species: Galeus melastomus FEMALE





1 IMMATURE IMMATURE



Uteri: enlarging.





3a


MATURE





4a POST‐LAYING MATURE

75



Uteri: enlarged.

4b REGENERATING MATURE

Species: Galeus melastomus MALE





1 IMMATURE IMMATURE








3a

CAPABLE

TO

REPRODUCE

MATURE




3b ACTIVE MATURE

76



4a REGRESSING MATURE

Lecithotrophic ‐ multiple oviparity ‐ Species: Galeus melastomus FEMALE

STAGE PHOTO

1

2

77

3a

3b

4a

78

4b

79

Species: Galeus melastomus MALE

STAGE PHOTO

1

2

3a

80

3b

4

81

Annex 6. Atlas for viviparous species

Lecithotrophic ‐ yolk‐sac viviparity ‐ Species: Etmopterus spinax FEMALE

PROPOSAL MATURATION

STATE STAGE PHOTO

Ovaries: small and whitish; undistinguishable ovarian follicles. Oviducal gland: often not visible. In some species a thickening of the uteri where the gland will develop may be visible Uteri: thread‐like and narrow.

IMMATURE 1 IMMATURE


DEVELOPING 2 IMMATURE


CAPABLE to REPRODUCE 3 MATURE


EARLY PREGNANCY 4a MATERNAL


MID PREGNANCY 4b MATERNAL


LATE PREGNANCY 4c MATERNAL


POST‐PARTUM 5 MATERNAL

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Ovaries: large with small and medium sized yolked follicles. Pre‐ovulatory follicles absent. Atretic follicles may be present. Oviducal glands: fully developed but may be reduced in size. Uteri: enlarged.

REGENERATING 6 MATURE

Species: Etmopterus spinax MALE

PROPOSAL MATURATION

STATE STAGE MATURITY PHOTO

Claspers: flexible, non calcified and usually shorter than pelvic fins. Testes: small and undeveloped Ducts: straight and thread‐like.

IMMATURE 1 IMMATURE




CAPABLE

TO

REPRODUCE

3a MATURE


ACTIVE 3b MATURE


REGRESSING 4 MATURE not available

83

Lecithotrophic ‐ yolk‐sac viviparity ‐ Species: Etmopterus spinax FEMALE

STAGE PHOTO

1

2

84

3

4a

4b

85

4c

5

6

86

Species: Etmepterus spinax MALE

STAGE PHOTO

1

2

87

3a

3b

4 not available

88

Lecithotrophic ‐ yolk‐sac viviparity ‐ Species: Somniosus rostratus FEMALE

PROPOSAL MATURATION



IMMATURE 1 IMMATURE








MID PREGNANCY 4b MATERNAL



89





90

Species: Somniosus rostratus MALE

PROPOSAL MATURATION



IMMATURE 1 IMMATURE not available




CAPABLE

TO

REPRODUCE

3a MATURE


ACTIVE 3b MATURE not available



91

Lecithotrophic ‐ yolk‐sac viviparity ‐ Species: Somniosus rostratus FEMALE

STAGE PHOTO

1

2

92

3

4a

4b

93

4c

5

6

Species: Somniosus rostratus MALE

PHOTO

94

Not available

1

2

3a not available

3b not available

4 not available

95

Matrotrophic – viviparity: FEMALE

PROPOSAL MATURATION



IMMATURE 1 IMMATURE








MID PREGNANCY 4b MATERNAL not available



Cetorhinus maximum

Myliobatis aquila

Deania calcea

Carcharhinus plumbeus


96





Deania calcea

Mustelus mustelus

97

MALE

PROPOSAL MATURATION



IMMATURE 1 IMMATURE




CAPABLE

TO

REPRODUCE

3a MATURE


ACTIVE 3b MATURE



Mustelus mustelus

Mustelus mustelus

Alopias vulpinus

Cetorhinus maximum

98

Matrotrophic ‐ viviparity: FEMALE

STAGE PHOTO

1

2

Cetorhinus maximum

Myliobatis aquila

99

3

4a

4b not available

Deania calcea


100

4c

5

6


Deania calcea

Mustelus mustelus

101

MALE

STAGE PHOTO

1

2

Mustelus mustelus

Mustelus mustelus

102

3a

3b

4 not available

Alopias vulpinus

Cetorhinus maximum

ICES WKMSEL REPORT 2012

Annex 7: WEBGR contact to upload photo reference library

WebGR is a European project that aims to develop Open Source software to support studies of fish growth and reproduction. In particular it promotes the use of online services to organise calibration workshops.

Scientists, who read otoliths to identify the ages of individual fish, have carried out calibration workshops for many years, to fine-tune their interpretation of the ages of fish within individual stocks or species. Calibration workshops have also recently been extended to cover the identification of fish gonad maturity stages. In general WebGR can be applied to all situations, where individual scientists need to discuss the interpretation of a protocol, for the identification of the status of biological mate-rial.

The consortium is constituted by: Portuguese Institute for the Sea and Atmosphere (IPMA) – Consortium leader, The Agri-Food and Biosciences Institute (UK), AZTI Tecnalia Foundation (Spain), Federal Agency for Agriculture and Food (Germany), Johann Heinrich von Thünen Institute (Germany), Hellenic Centre for Marine Re-search (Greece), Instituto Español de Oceanografia (Spain), Institut français de re-cherche pour l’exploitation de la mer (France), Institute for Marine Resources and Ecosystem Studies (The Netherlands), Institute of Marine Research (Norway), Swe-dish Board of Fisheries (Sweden), Italian Society for Marine Biology (Italy).

Note that all information uploaded will be under a Creative Commons Attribution-Share Alike 3.0 Unported License (link this to Creative Commons Attribution-Share Alike 3.0 Unported License.

For more information please visit http://webgr.wiki.azti.es.

http://creativecommons.org/licenses/by-sa/3.0/

http://creativecommons.org/licenses/by-sa/3.0/

http://webgr.wiki.azti.es/

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Annex 8. Working documents presented by corresponding partici-pants

Application of the WKMSEL scales during the MEDITS survey conducted off the Balearic Islands (GSA 05)

by Gabriel Morey

This report contains the description of the activities related to the assessment of ma-turity of elasmobranchs during the MEDITS survey conducted off the Balearic Islands (GSA 05) between May 29 and June 12 2012 by the COB-IEO (Centre Oceanogràfic de Balears - Spanish Oceanographic Institute).

Study area

The survey was carried out over shelf and upper slope trawlable bottoms around Mallorca and Menorca islands, at a depth range of 50-800 m. A total of 50 hauls were performed at 4 depth strata: B (51-100 m; n=20), C (101-200 m; n= 14), D (201-500 m; n=8) and E (501-800 m; n=8).

Species recorded during the survey

18 elasmobranch species (7 sharks and 11 batoids) plus a chimaera were reported during the survey. A total of 1588 specimens were captured (1169 sharks, 418 batoids and 1 chimaera). Among sharks, the most common species were Scyliorhinus canicula and Galeus melatomus, representing 51% and 43% of the sharks, respectively. For ba-toids, the most important species were Raja clavata and Raja polystigma, accounting for 38% and 25%, respectively.

Table 1. Number of individuals by sex for all the Chondrychthyan species reported during the survey, and depth range where they were captured.

Sex Depth range (m)

Species Females Males Unsexed Total Min. Max.

Dalatias licha 1 1 743

Etmopterus spinax 8 11 19 421 744

Galeus melastomus 191 316 1 508 356 744

Mustelus mustelus 1 1 2 57 57

Oxynotus centrina 1 1 98

Scyliorhinus canicula 253 362 615 51 398

Squalus blainvillei 9 13 1 23 172 744

Dasyatis pastinaca 5 4 9 56 56

Dipturus oxyrinchus 13 7 20 160 421

Myliobatis aquila 6 4 10 51 147

Leucoraja naevus 33 20 53 69 252

Raja brachyura 2 2 57 57

Raja clavata 88 70 158 70 421

Raja miraletus 24 16 40 56 150

Raja polystigma 54 49 103 58 398

Raja radula 15 4 19 51 147

Rostroraja alba 1 1 2 77 251

Torpedo marmorata 1 1 2 51 77

Chimaera monstrosa 1 1 601

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Sampling

All elasmobranch species were individually sampled as indicated in Table 2. Because the large number of G. melastomus and S. canicula individuals, these two species were measured to the lowest 0.5 cm, and both eviscerated, gonads and liver weights were not taken. For batoids, further to total length, the disc width was also taken in order to determine length-length relationships or weight-length relationships through lengths other than TL, as batoids are usually landed with tails cut off.

Table 2. Measures taken for the different species or groups. TL: total length; LP: length to pelvic fins; DW: disc width; Wt: total weight; We: eviscerated weight; Wg: gonads weight; Wl: liver weight.

Sex Maturity TL LP DW Wt We Wg Wl

Galeus melastomus

Scyliorhinus canicula

Rest of shark species

Dasyatis pastinaca Myliobatis aquila Rest of batoid species

Maturity assessment

The maturity scale officially used during the survey was that previously used in the MEDITS program, this is, stages 1 to 4 both for males and females as following:

- Stage 1: Immature

- Stage 2: Maturing

- Stage 3: Mature / Spawning

- Stage 4: Post-spawning / Resting

This scale was applied to G. melastomus and S. canicula. Because the large amount of individuals of both species, they were sampled by part of the staff not familiarized with the maturity scale resulting from the WKMSEL 2010.

Whereas, the rest of the elasmobranchs were sampled by other part of the staff that had been previously studying the maturity scale proposed at the WKMSEL 2010. The detailed maturity scales (including descriptions of gonads and claspers) and macro-scopic gonad reference photos, as appearing on the WKMSEL report 2010, were available to the staff and consulted continually during all the sampling.

Due to that the standardized database used during the survey did not allow to in-clude subdivisions of maturity stages, an additional field was added for doing so, but maintaining the former one where stages 1 to 4 (without subdivisions) were reported.

Photos of gonads belonging to 35 individuals, with the corresponding information (species, sex, TL and maturity stage assessed) were contributed to the 2012 WKMSEL with the purpose of checking the accuracy when assessing maturity stages during the survey.


Annex 9:Maturity scales proposed by WKMSEL2 (2012)

Oviparous elasmobranchs (skates and sharks) - FEMALES


IMMATURE

1 IMMATURE (Immature)

Ovaries: small and whitish. Undistinguishable ovarian follicles. Oviducal gland: often not visible. In some species a thickening of the uteri where the gland will develop may be visible. Uteri: thread-like and narrow.

2 DEVELOPING (Immature)

Ovaries: follicles of different stages of development. Some small and medium sized yolked follicles may be present. Oviducal gland: distinguishable and developing. Uteri: enlarging.

MATURE

3a

CAPABLE TO REPRODUCE (mature)

Ovaries: presence of large yolked follicles ready to be ovulated. Oviducal glands: fully developed. Uteri: fully developed.

3b EGG-LAYING (mature)

Uteri: presence of egg capsules.

4a POST-LAYING (mature)

Ovaries: flaccid with few follicles of different sizes. Few large vitellogenic follicles entering atresia. POFs and atretic follicles may be visible. Oviducal glands: fully developed but may be reduced in size. Uteri: enlarged and flaccid.

4b REGENERATING (mature)

Ovaries: large with small and medium sized yolked follicles. Pre-ovulatory follicles absent. Oviducal glands: fully developed but may be reduced in size. Uteri: enlarged.


Oviparous elasmobranchs (skates and sharks) - MALES


IMMATURE

1 IMMATURE

Claspers: flexible, non-calcified and shorter than pelvic fins. Testes: small and undeveloped (in skates, sometimes with visible lobules). Ducts: straight and thread-like.

2 DEVELOPING

Claspers: flexible, partially calcified and usually as long as or longer than pelvic fins.* Testes: developing (in skates, lobules clearly visible but not fully developed). Ducts: developing and beginning to coil.

MATURE

3a


Claspers: rigid, fully calcified, and longer than pelvic fins (in some sharks they may only be as long as the pelvic fins). Testes: fully developed (in skates, with fully formed lobules). Ducts: tightly coiled and filled with sperm.

3b ACTIVE

Claspers: similar to stage 3a, however with clasper glands dilated, sometimes swollen and reddish. Sperm may be present in clasper groove or glands. Testes: similar to stage 3a. Ducts: sperm observed inside (after a cut) or flowing out of the cloaca on pressure.

4 REGRESSING

Claspers: fully formed, similar to stage 3. Testes shrunken and flaccid, (in skates, with few visible lobules). On pressure sperm does not flow. Sperm ducts: empty and flaccid


Viviparous elasmobranchs (skates and sharks) - FEMALES

MATURITY/ MATERNITY

STAGE DESCRIPTION

IMMATURE

1 IMMATURE

Ovaries: small and whitish; undistinguishable ovarian follicles. Oviducal gland: often not visible. In some species a thickening of the uteri where the gland will develop may be visible Uteri: thread-like and narrow.

2 DEVELOPING


MATURE 3 CAPABLE to REPRODUCE


MATERNAL

4a EARLY PREGNANCY


4b MID PREGNANCY


4c LATE PREGNANCY


5 POST-PARTUM


MATURE 6 REGENERATING

Ovaries: large with small and medium sized yolked follicles. Pre-ovulatory follicles absent. Atretic follicles may be present. Oviducal glands: fully developed but may be reduced in size. Uteri: enlarged.


Viviparous elasmobranchs (skates and sharks) - MALES


IMMATURE

1 IMMATURE

Claspers: flexible, non-calcified and usually shorter than pelvic fins. Testes: small and undeveloped Ducts: straight and thread-like.

2 DEVELOPING


MATURE



3b ACTIVE


4 REGRESSING
