Reproductive biology of the orangespotted grouper Epinephelus coioides (Pisces: Serranidae) from coastal Kenya

By Simon Were Agembe REPORT NO: WIOMSA/MARG-I/2008 14

Abstract The fisheries of groupers have declined world wide mainly due fishing pressure on coral reef fishes. Although data on groupers reproductive biology are important for management of fisheries, such data are lacking in coastal Kenya. This study therefore aimed to determine i) the size at first maturity of the species ii) the spawning time of the species at both the small (lunar cycle) and large (monthly cycle) temporal-scales iii) the sex ratio of the commercially important orange-spotted grouper, Epinephelus coioides at Shimoni on south coastal Kenya. Data were obtained at Shimoni from July 2007 to September 2008. Specimens were measured for weights, lengths and dissected, sexed and assigned maturity stages. The length-weight relationship results for the orange-spotted grouper, Epinephelus coioides, calculated was Wp = 0.011745 TL3.04. Sex ratio was 1:27.5 in favour of females. A chi-squared goodness of fit test showed that sex ratio was significantly different from a 1:1 ratio (2 = 49.281, df = 1, p = 0.000). The orangespotted grouper, Epinephelus coioides, females had a length frequency distribution that was skewed to the left indicating dominance of young fishes in the catch. It was concluded that the sex ratio of the orange-spotted grouper, Epinephelus coioides, showed a deviation from unity sex ratio in favour of females. The length-frequency distributions showed high proportion of fishes were caught before reaching sexual maturity. The presence of high proportions of immature female fishes in the fishery suggested growth overfishing and spawning was found to be synchronized to lunar phase. Epinephelus coioides spawns during new moon. It was recommended that there is need for the Department of Fisheries, GoK, to restrict exploitation of Epinephelus coioides during new moon phase. This lunar phase is the spawning window for this species, thus such restrictions will provide conservation of their spawning stock biomass. There is also need for studies to determine the factors that have led to the decreased abundance of the orange-spotted grouper, Epinephelus coioides. These will include determination of MSY, variation of spawning aggregations together with estimate of grouper densities.

Acknowledgements I wish to thank WIOMSA for funding this research through the MARG I Grant. I am also grateful to the Director and the entire management of KMFRI for support that I received during the execution of this research. Mr. Boaz Orembo, Mr. Dickson Odongo, James Gonda, Bernard Ogongo, Daniel Ocharo and Keneth Omondi assisted in data collection.

Table of Contents Abstract ............................................................................................................................... 2 Acknowledgements............................................................................................................. 3 Table of Contents................................................................................................................ 4 List of Figures ..................................................................................................................... 5 1.0 Introduction................................................................................................................... 6 2.0 Materials and Methods.................................................................................................. 9 2.1 Study site................................................................................................................... 9 2.2: Data collection ....................................................................................................... 11 2.3 Histological Techniques.......................................................................................... 12 2.4 Data Analysis .......................................................................................................... 12 3.0 Results......................................................................................................................... 13 3.1 Length-weight relationship ..................................................................................... 13 3.2 Sex ratio and length-frequency distribution............................................................ 13 3.3 Sex at first maturity................................................................................................. 13 3.4 Gonad maturity stages............................................................................................. 14 3.5 Proportion of mature fish and lunar spawning........................................................ 16 4.0 Discussion ................................................................................................................... 17 5.0 Conclusions................................................................................................................. 19 5.1 Recommendations................................................................................................... 19 6.0 References................................................................................................................... 20

List of Figures Figure 1: Map of the study area showing Shimoni landing site on the South coast, Kenya . 10

Figure 2: Length-frequency distribution for female orange-spotted grouper Epinephelus coioides (n = 114) landed in Shimoni. Vertical dashed lines indicate size at first maturity . 14

Figure 3: Temporal variations in proportion of mature and immature fishes in daily catches of Epinephelus coioides landed in Shimoni with lunar phase transposed on the data. () indicates new moon while () indicates full moon phase . 17

1.0 Introduction Groupers and hinds (Teleostei, Serranidae) form a valuable component of catches of reef fishes in Kenya (Kaunda-Arara et al., 2003) and the rest of Eastern Africa (Nzioka, 1979; Jiddawi & Stanley, 1999). Catch rates and species diversity of reef fishes in Kenya have declined ((Kaunda-Arara et al., 2003) due to overexploitation. Groupers are long lived slow growing fishes, a trait that makes them vulnerable to over exploitation (Sadovy, 2000). Knowledge of their reproductive biology is important in understanding their population dynamics and for regulation of exploitation. The orange-spotted grouper, Epinephelus coioides Hamilton, 1882 is one of the most common groupers found in tropical estuaries and coastal reefs (Prokop, 2002). The Orange-spotted grouper is economically important and expensive fish in the markets (Heemstra & Randall, 1993), it is commonly taken in juvenile form in some parts of the world for grow-out mariculture (Sadovy, 2000). For this significant commercial species, little is known of its biology in the Western Indian Ocean (WIO) region. This species is known to form spawning aggregations in parts of the pacific and Caribbean waters (Colin et al., 2003), increasing its vulnerability to overexploitation in countries where biological data and exploitation status is scarce like in this region. This study therefore aimed to determine i) the size at first maturity of the species ii) the spawning time of the species at both the small (lunar cycle) and large (monthly cycle) temporal-scales and iii) the sex ratio of the orangespotted grouper, Epinephelus coioides at Shimoni on south coast, Kenya. 1.1 Literature Review The Serranid sub family Epinephelinae is widespread in both tropical and subtropical seas (Heemstra & Randall, 1993) where they are heavily fished (Kaunda-Arara et al., 2003). About 90% of the worlds harvest of marine food is derived from artisanal 6

fisheries, and groupers (Serranidae) are a major component (Heemstra & Randall, 1993). Among the groupers (sub family Epinephelinae) protogyny is reported as the most common mode of reproduction (Rhodes & Sadovy, 2002) while in some males are subsequently absent until the cohort reaches the age of 10 and the cohort sex ratio is not equal until age 15 (Shapiro, 1987 b; Manooch, 1987). This mode of reproduction presents problems for fishery management (Tupper, 1999), because the effects of fishing on protogynous populations are exceedingly difficult to measure without complete information on their reproductive patterns, sex ratio and some other biological aspects of fish populations (Johannes, 1978; Koenig et al., 1996; Coleman et al., 2000; Rhodes & Sadovy, 2002; Sadovy & Domeier, 2005; Pears et al., 2007).

In the Indo-pacific region little data exist on the biology of groupers except general observations (Nzioka, 1979) interview based spawning aggregations (Samoilys et al., 2004; Robinson et al., 2004) but fish are increasingly threatened with overexploitation (Kaunda-Arara et al., 2005; Jiddawi & Stanley, 1999). In coastal Kenya the fisheries have shifted from hand-line capture for the large groupers to drive net, seine (Kaunda-Arara et al., 2003) while most are caught primarily using traps, seines, and bottom set lines, and is sought after by spear fishers.

The orange spotted grouper, Epinephelus coioides (Pisces: Serranidae) is one of the most important food fishes globally. It is believed to be a protogynous hermaphrodite in which most if not all individuals begin life as females and subsequently becoming males (AbuHakima, 1987; Huntsman & Schaaf, 1994; Liu & Sadovy, 2004; Grandcourt et al., 2005). 7

It is reported that selective removal by a fishery of larger sized individual males in protogynous species may result in sex ratio imbalances that could lead to sperm limitation and reduced fertilization rates (Rhodes & Sadovy, 2002). The determinations of the sex ratio and changes in maturity stages during the year are of considerable importance in building knowledge of the general biology of an exploited stock (Taylor, 1984). These determinations provide basic knowledge of the reproductive biology of a stock. The information derived from these analyses may be used in ascertaining the size at which fish attain sexual maturity, the time and place of spawning and the duration of the cycle from the beginning of the development of the ovary to the final release of eggs. These types of data have several practical uses, in assessing the optimum age of first capture of a species and the time and place of spawning, to plan fishing tactics because many species are easiest to catch when they congregate to spawn. Fishing therefore reduce the reproductive potential of stock by possibly removing spawner biomass thus reducing average fish size (Fulton et al., 1999) since this is done with no requisite biological examination of specimens (Domeier et al., 2002; Church pers.com.). Reports indicate that the orange-spotted grouper, Epinephelus coioides, is categorized as Near Threatened (IUCN) (SCRFA, 2007). And large losses in mangroves, key habitat for young Epinephelus coioides in many areas aggravate the problem further. 1.2 Objectives The Overall objective of this project was to provide initial data on the reproductive biology of the commercially important orange-spotted grouper, Epinephelus coioides from coastal Kenya that can be used for scientific management of the stocks. The specific objectives of the project were: 8

i). to determine the size at first maturity of the species ii). to determine the spawning time of the species at both the small (lunar cycle) and large (monthly cycle) temporal-scales. iii). to determine the sex ratio of the species landed in fisher catches

2.0 Materials and Methods 2.1 Study site The study was undertaken in three landing beaches on the southern coast of Kenya. The beaches were Msambweni, Shimoni and Vanga (Fig. 1). Shimoni is located on the Pemba channel, it has three fishing villages, two on the mainland and one in the adjacent Wasini Island (Ochiewo, 2004). Part of its traditional fishing ground was converted into a marine protected area, the Kisite-Mpunguti Marine National Park and Reserve, which is a major tourist attraction on the south coast of Kenya. The beach was chosen because it one of the most active on the south coast of Kenya. Additionally, it is off the more pristine reefs on the Kenyan coast, which likely make the site to have relatively higher grouper landings. The fishing activities off this landing area are concentrated within the reef lagoons as fishermen hardly venture beyond the outer reef area because of the fishing crafts they use.

Figure 1: Map of the study area showing Shimoni landing site on the south coast, Kenya

10

2.2: Data collection Sampling was done at Shimoni on the Kenyan Coast; Epinephelus coioides samples were trapped using the traditional Dema traps (Kaunda-Arara & Ntiba, 1997) widely used in coastal East Africa. Sampling was carried for a period of 12 months. During each month sampling was carried for a period of 14 continuous days. The Dema traps were set within the shallow reef lagoons next to coral heads where groupers were expected to hide. The groupers were baited with animal tissues (e.g. mashed gastropods etc). The traps were inspected after 24 hours of soaking. On each fishing day, the traps were overhauled with the help of experienced fishers on a fishing canoe, contents inspected and samples of Epinephelus coioides sorted. Samples were also collected from other gears including spear gun, hand lines and long liners which landed the species. This was due to low or no samples from traps. The total (TL) and standard lengths (SL) of fish were measured on a measuring board to the nearest 0.01cm and wet body weight on a top-loading balance to nearest 0.1 g. Fish specimens were then dissected, sex determined and gonad maturity stages recorded where stages III onwards were considered to be mature for males and females respectively. The whole gonad was weighed to the nearest 0.01 g on an analytical balance. For females, the size at first maturity was determined by calculating the proportion of mature individuals in each size class. The size at which 50% of individuals were mature was taken as the size at which this fish species reach maturity for the first time.

11

2.3 Histological Techniques Portions of some gonads at different stages of maturation were fixed in formalin, acetic acid and calcium chloride (FAACC) for later histological sectioning in the laboratory to confirm the sexes. Tissues for sectioning were embedded in paraffin wax, sectioned at 7 m, stained with Harris Haematoxylin and counterstained with eosin. Ovaries were assigned a stage on the basis of the most advanced stage of oocyte development (West, 1990).

2.4 Data Analysis Length-weight relationships were determined for grouper species using the equation: Log10 W = log10 a + b log10 TL, where W is the body weight, TL is the total length, a is the intercept, and b is the slope of regression line (Wooton, 1990). This relationship was estimated for fifteen species that had 15 or more specimens. The slope of the lengthweight relationship for this species was tested for significant difference from 3.0 following the one tailed t-test procedure (Zar, 1999), in order to determine whether growth is isometric or allometric. Length frequency distributions were used to describe the size structure of the common grouper species. The differences in sex ratio were analysed for significant divergence from expected 1:1 ratio by using a Chi-square (2) goodness of fit test (Zar, 1999). Reproductive activity was analyzed on a lunar temporal scale by relating the relative proportion (%) of immature (stages I-II) and mature fishes (stage III and

above) in the daily samples for 4 grouper species with the lunar phase. Statistical data analyses were done using MINITAB release 13, MS-Excel and Sigma plot computer packages. The non parametric Chi-square test (Zar, 1999) was used to test for significant 12

deviation from the expected male to female ratio of 1:1. For all statistical tests significance was determined at = 0.05.

3.0 Results 3.1 Length-weight relationship The length-weight relationship results for the orange-spotted grouper, Epinephelus coioides, calculated from the data was Wp = 0.011745 TL3.04. The t-test performed on the slope showed no significant difference from the value 3 an indication of isometric growth.

3.2 Sex ratio and length-frequency distribution A total of 114 specimens of the orange-spotted grouper, Epinephelus coioides, were sexed of which 110 were females and 4 were male, giving a sex ratio of 1:27.5 in favour of females. A chi-squared goodness of fit test showed that the ratio was significantly different from a 1:1 ratio (2 = 49.281, df = 1, p = 0.000). The orange-spotted grouper, Epinephelus coioides, females had a length frequency distribution that was skewed to the left indicating dominance of young fishes. The modal frequency for this species was 30.0-35.0 cm size groups, fishes of sizes > 50.0 cm were few in the samples (Fig. 2). The mean size for this species was 46.3 18.55. Members of this species mature as females.

3.3 Sex at first maturity The determination of the average size at which 50 % of Epinephelus coioides males and females attain first sexual maturity was not possible from the data collected during this 13

study. However, the size at first maturity on this species was sourced from the global database and is shown in dashed lines in Figure 2.

20 18

Number of individuals

16 14 12 10 8 6 4 2 0 100-105 20-25 25-30 30-35 35-40 40-45 45-50 50-55 55-60 60-65 65-70 70-75 75-80 80-85 85-90 90-95 95-100

Size classs (cm)

Figure 2: Length-frequency distribution for female orange-spotted grouper Epinephelus coioides (n = 114) landed in Shimoni. Vertical dashed lines indicate size at first maturity.

3.4 Gonad maturity stages The description of maturity stages for gonads in both males and females of the orange spotted grouper Epinephelus coioides is given in Table 1 and 2. Both macroscopic and microscopic appearances of the ovary are shown for females. For males only macroscopic appearance is given. 14

Table 1: Description of macroscopic maturity stages of Epinephelus coioides gonads based on the modified maturity schemes from Rhodes & Sadovy (2002). a) OVARIES Maturity Stage I = Immature Macroscopic features Ovary small, strand-like, compact, pink or cream; oocytes (eggs) not clearly distinct; not obviously different from immature or inactive males II = Maturing Ovary relatively small but rounded, less strand-like in appearance, grayish with thickened gonad wall; eggs not clearly distinct and small ; Not clearly different from mature males prior to the development of yolk within the eggs III= Mature/active Ovary large and grayish with transparent gonad wall; large yolky eggs becoming clearly visible and tightly packed IV-V=Mature/ ripe Ovary relatively large, clear, watery (hydrated) eggs visible through wall; typical of individuals just prior to spawning; egg release possible with application of light abdominal pressure VI = Post-spawn b) TESTES I=Immature/ Inactive II = Maturing Ovary flaccid with obvious capillaries (small blood vessels); few eggs visible Testes not obviously different from immature females (see the description of immature females) Early Male II individuals are indistinguishable from Female II. Single blood vessel on ventral side. No sperm extricable when pressed lightly

III= Mature/active Testes expanding and becoming rounded and large; grayish in appearance; early maturing individuals are not clearly different from maturing females until milt (sperm) becomes evident in the sperm sinus along the gonad wall IV-V=Mature/ ripe Testes large and white with sperm visible in sinuses along the gonad wall; milt release with light abdominal pressure VI = Post-spawn Testes flaccid and bloody; sperm release still possible on application of abdominal pressure

15

Table 2: Description of microscopic maturity stages of Epinephelus coioides ovaries based on the modified gonad maturity schemes from Moe (1969). Maturity stage Immature Mature resting Mature active Mature running Spent Ovaries Oocytes exist only as oogonia and stages 1 and 2, unless otherwise stated in the text. Atretic bodies do not exist Ooocytes exist as oogonia and stages 1 and 2. Atretic bodies located generally central to the lamellae. Ovary is undergoing vitellogenesis and oocytes are predominantly at stage stages 3 Hydrated oocytes, post-ovulatory follicles and atretic oocytes The gonad structure is greatly disrupted, empty follicles are evident and the density of tissue in lamellae is sparse

3.5 Proportion of mature fish and lunar spawning The results on percent proportion of mature and immature fishes showed there were more immature fishes in the catch (Fig. 3). The catch comprised mainly of the females. In the month of July there were distinct peaks for both mature and immature fishes. The temporal variations in proportions of mature and immature orange-spotted grouper, Epinephelus coioides, landed in Shimoni is shown in Figure 3. In this species proportionately mature individuals ( 25 %) were caught on day 193 coinciding with the new moon phase. A small proportion ( 15 %) of mature individuals was observed on days 169, 197 and 201 all corresponding with full moon lunar phase. A large proportion (30 %) of immature individuals was caught on day 192 corresponding to the new moon phase. Other distinct peaks in occurrence of immature E. coioides were observed on days 165 and 194 which coincided with the full moon lunar phases.

16

Mature fish Immature fish

Lunar phase

35 30 % Proportion 25 20 15 10 5 0 157 159 161 163 165 167 169 190 192 194 196 198 200 202 Day of year

Figure 3: Temporal variations in proportion of mature and immature fishes in daily catches of Epinephelus coioides landed in Shimoni with lunar phase transposed on the data. () indicates new moon while () indicates full moon phase.

4.0 Discussion A skewed sex-ratio in favour of females in groupers has also been reported by several workers (Shapiro 1987 b; Rhodes & Sadovy, 2002; Grandcourt et al., 2005; Rhodes & Tupper, 2007). Sadovy (1996) and Rhodes and Tupper (2007) noted that biased sex-ratios of groupers is a typical characteristic of protogynous hermaphrodites. However, Jones et al., (2004) suggested biased sex ratios may be attributed to differences in spatial distribution and schooling behaviour between sexes. 17

The results of length-frequency distributions analyses for females of orange-spotted grouper, Epinephelus coioides, indicated that mature fishes are few in the landings while immature fish comprised large proportion of the catches. The modal size class for the orange-spotted grouper species (30-35 cm) is below the reported size at first maturity (Froese et al., 2003) for the orange-spotted grouper, Epinephelus coioides. The higher proportion of smaller fish in catches suggests that the orange-spotted-grouper fisheries may be experiencing growth overfishing (Thompson & Munro, 1983), which occurs when large fish are being harvested usually because of excessive effort thereby reducing the average size in the population (McClanahan et al., 2008). Although no individual fish with both ovarian and testicular tissues were observed in my histological studies, the predominance of females in the smaller size classes was taken to suggest Epinephelus coioides matures as females then changes to males.

The results of this study suggested that spawning in orange-spotted-grouper, Epinephelus coioides is influenced by lunar phase. For example, spawning in Epinephelus coioides coincided with the new moon phase (Fig. 3). These findings are in agreement with results on spawning in Serranidae occurring during specific lunar periods as reviewed by Johannes (1978). Elsewhere Epinephelus striatus is reported to spawn around the new moon in the Bahamas (Smith, 1972) while Epinephelus merra and E. striatus are reported to spawn at the full moon lunar phase at the Society Islands (Randall, 1964) and off Belize (Johannes, 1978) respectively. The results in this study are contrary to most findings of reef fishes in the NEM season in Kenya (Nzioka, 1979; Kaunda-Arara & 18

Ntiba, 1997). However, it may be that the lunar based spawning of groupers makes it difficult to detect spawning on a larger monthly scale applied in other studies.

5.0 Conclusions The sex ratio of the orange-spotted grouper, Epinephelus coioides, showed a deviation from unity sex ratio in favour of females. The length-frequency distributions of this species showed high proportion of fishes were caught before reaching sexual maturity. The presence of high proportions of immature female fishes in the fishery suggested growth overfishing. Spawning was found to be synchronized to lunar phase. Epinephelus coioides spawns during new moon.

5.1 Recommendations From the results of this study the following recommendations are made: There is need for the Department of Fisheries, GoK, to restrict exploitation of Epinephelus coioides during new moon phase. This lunar phase is the spawning window for this species, thus such restrictions will provide conservation of their spawning stock biomass. There is need for studies to determine the factors that have led to the decreased abundance of the orange-spotted grouper, Epinephelus coioides. These will include determination of MSY, variation of spawning aggregations together with estimate of grouper densities.

19

6.0 References Abu-Hakima, R. (1987) Aspects of the reproductive biology of the grouper, Epinephelus tauvina (Forskl), in Kuwait waters. Journal of fish biology 30:213-222 Coleman, F.C; Koening, C.C & Collins, L.A. (1996) Reproductive styles of shallow water groupers (Pisces: Serranidae) in the eastern Gulf of Mexico and the consequences of fishing spawning aggregations. Environmental Biology of fishes 4747:129-141 Colin, P.L.; Sadovy, Y.J. & Domeier, M.L. (2003) Manual for the study and conservation of reef fish spawning aggregations society for the conservation of the reef fish aggregation special publication No.1 (version 1.0). Pp1-98 + iii Domeier, M.L.; Colin, P.L.; Donaldson, T.J., Heyman, W.D., Pet, J.S., Russell, M., Sadovy, Y., Samoilys, M.A., Smith, A., Yeeting, B.M. & Smith, S. (2002) Transforming coral reef conservation: Reef fish spawning aggregation component working group report. 139pp Froese, R., Pauly, D. & Woodland, D. (2003) "FishBase" (On-line). FishBase World Wide Web electronic publication. Accessed at http://www.fishbase.org/ Fulton, E.; Kault, D.; Mapstone, B. & Sheaves, M. (1999) Spawning season influences on commercial catch rates: computer simulations and Plectropomus leopardus, a case in point. Canadian journal of fisheries and aquatic science 56: 1096-1108 Grandcourt, E.M.; Al Abdessalaam, T.Z.; Francis, F.; Al Shamsi, A.T (2005) Population biology and assessment of the orange-spotted grouper, Epinephelus coioides (Hamilton, 1822), in the southern Arabian Gulf. Fisheries Research 74: 55-68 Heemstra, P.L. & Randall, J.E. (1993) FAO species catalogue.vol.19.Groupers of the world. Family Serranidae, Subfamily Epinephelinae. An annotated and illustrated 20

catalogue of the grouper and lyretail species known to date .FAO Fisheries Synopsis,vol.125,no.16,FAO,Rome.382p Huntsman, G.R. & Schaaf, W.E. (1994) Simulations of the impact of fishing on reproduction of a protogynous grouper, the graysby. North American Journal of Fisheries Management 14: 41-52 Jiddawi, N.S. & Stanley, R.D. (1999) Fisheries stock assessment in the traditional fisheries sector: The Information needs. In Proceedings of the National Workshop on the Artisanal Fisheries Sector, Zanzibar, September 2224, 1997 N. S. Jiddawi & R.D. Stanley (Editors) Zanzibar, Tanzania Johannes, R.E. (1978) Reproductive strategies of coastal marine fishes in tropics. Environmental fish biology 3 (1) : 65-85 Jones, G.P., McCormick, M.I., Srinivasan, M., Eagle, J.V. (2004) Coral decline threatens fish biodiversity in marine reserves. Proceedings of the National Academy of Science 101: 82518253 Kaunda-Arara, B. & Ntiba, M.J. (1997) The reproductive biology of Lutjanus fulviflamma (Forssskl, 1775) (Pisces: Lutjanidae) in Kenyan inshore marine waters Hydrobiologia 353:153-160 Kaunda-Arara, B. (2005) Preliminary investigations into spawning aggregations in coastal Kenya. A technical report prepared for IUCN, Nairobi. 27pp Kaunda-Arara, B., Rose, G.A., Muchiri, M.S. & Kaka, R. (2003) Long-term trends in coral reef fish yields and exploitation rates of commercial species from coastal Kenya. Western India Ocean Journal of Marine Science 2: 105-116

21

Koening, C.C; Coleman, F.C; Collins, L.A; Sadovy, Y & Colin, P.L. (1996) Reproduction in gag (Mycteroperca microlepis) (Pisces: Serranidae) in the Eastern Gulf of Mexico and the consequences of fishing spawning aggregations. ICLARM conference proceedings 48: 307-323 Liu, M. & Sadovy. Y. (2004) Early gonadal development and primary males in the protogynous Epinepheline, Cephalopholis boenak. Journal of fish Biology 65: 9871002 Manooch. C.S. III. (1987) Age and growth of snappers pages 329-373 in J.J. Polovina and S. Ralston, eds. Tropical snappers and groupers: biology and fisheries management, Westview Press. Boulder Colorado McClanahan, T.R., Hicks, C.C. & Darling, E.J., (2008) Malthusian overfishing and efforts to overcome it on Kenyan coral reefs. Ecological Applications 18 (6): 1516-1529 Moe, M.A. Jr. (1969) Biology of the red grouper, Epinephelus morio (Valenciennes) from eastern Gulf of Mexico. Professional paper series marine laboratory Florida 10:195. Nzioka, R.M. (1979) Observations on the spawning seasons of East African reef fishes. Journal Fish Biology 14: 239-342 Ochiewo, J. (2004) Changing fisheries practices and their socioeconomic implications in South Coast Kenya. Ocean and Coastal Management 47: 389-408 Pears, R.J.; Choat, J.H.; Mapstone, B.D. & Beggs, G.A. (2007) Reproductive biology of large, aggregation spawning Serranid, Epinephelus fuscoguttatus

(Forsskl):management implications. Journal of fish biology 71:795-817

22

Prokop, F. (2002) Australian Fish Guide. Australian Fishing Network, Victoria, Australia. 2nd reprint. Pp 224 Randall, J.E. (1964) Notes on the groupers of Tahiti, with description of a new serranid fish genus. Pacific Science 18: 281296 Rhodes, K.L & Sadovy, Y. (2002) Reproduction in the camouflage grouper (Pisces: Serranidae) in Pohnpei, Federated States of Micronesia. Bulletin of Marine science 70(3): 851-869 Rhodes, K.L. & Tupper, M.H. (2007) A preliminary market-based analysis of the Pohnpei, Miconesia, grouper (Serranidae: Epinephelinae) fishery reveals unsustainable fishing practices. Coral Reefs 26: 335-344 Robinson, J., Isidore, M., Marguerite, M.A., hman, M.C.,& Payet .R.J. (2004) Spatial and temporal distribution of reef fish spawning aggregations in the Seychelles An interview-based survey of artisanal fishers. Western Indian Ocean Journal of Marine Science 3 (1):63-69 Sadovy, Y. (2000) Regional survey for fry/fingerling supply and current practices for grouper mariculture: evaluating current status and long-term prospects for grouper mariculture in South East Asia. Asia-Pacific Economic Cooperation Completion Report. 102 p. + App. + Plates http://www.enaca.org/grouper/Project/Fry/survey.htm Sadovy, Y.J & Domeier, M.L. (2005) Perplexing problems of sexual patterns in the fish genus Paralabrax (Serranidae; Serraninae).Journal of zoology 267:121-133 Sadovy, Y.J. (1996) Reproduction of reef fish. In Polunin, N.V.C.; Roberts, C.M (eds). Reef fisheries. Chapman and Hall. Fish and fisheries series pp 16-53 23

Samoilys, M.A; Church J.E; Kaunda-Arara, B.; Kamukuru, A & Jiddawi, N. (2004) Preliminary findings on spawning aggregations of reef fishers in East Africa. Proceedings for the 10th International Coral Reef Symposium, Okinawa, Japan SCRFA. (2007) About threatened and spawning aggregations. http://www.scrfa.org/serve. 10th July 2007 Shapiro, D.Y. (1987 b) Reproduction in groupers .Pages 295-327 in J.J. Polovina and S. Ralston, eds. Tropical snappers and groupers: biology and fisheries management, Westview Press. Boulder Colorado Smith, C.L., (1972) A spawning aggregation of Nassau grouper, Epinephelus straitus (Bloch). Transactions of the American Fisheries society 257-261 Taylor, M.H. (1984) Lunar synchronization of fish reproduction. Transactions of the American Fisheries society 113:484-494 Thomson, R. & Munro, J. L. (1983) The biology, ecology and bionomics of the hinds and groupers, Serranidae. In: Munro, J.L. (ed.). Caribbean coral reef fishery resources (ICLARM Stud. Rev. 7), ICLARM, Manila, Philippines, pp. 94109 Tupper, M.H. (1999) A brief review of grouper reproductive biology and implications for management of the Gulf of Mexico gag groupers fisheries 8pp West, G. (1990) Methods of assessing ovarian development in fishes: a review. Australian Journal of Marine and Freshwater Research 41: 199222 Wooton, R.J. (1990) Ecology of teleost Fishes. Fish and Fisheries series. Chapman and Hall, New York 404 pp. Zar, J.H. (1999) Biostatistics Analysis. Prentice Hall, New Jersey

24