Quantitative measurement of reproductive output in the American oyster, Crassostrea virginica (Gmelin), using an enzyme-linked immunosorbent assay (ELISA)

Abstract. A quantitative gonadal index was developed for oysters, Crassostrea virginica (Gmelin. 1791), using polyclonal antibodies from eggs and sperm. Percoll used in the purification of oyster eggs and sperm greatly improved the purity of antigens compared to filtering the egg or sperm through a fine mesh only. The antigen-antibody reaction was tested with indirect sandwich ELISA using alkaline phosphatase-conjugated goat anti-rabbit igG as a secondary antibody. Rabbit anti-oyster egg IgG and anti-oyster sperm IgG initially exhibited a weak cross-reactivity over somatic tissue. Absoring with acetone-dried oyster tissue powder removed this cross-reactivity. Both antisera exhibited strong specific immunological reactions to oyster eggs or sperm respectively. The quantity of eggs or sperm was measured using ELISA and a quantitative gonadosomatic index (dry wt of egg or sperm/dry wt oyster) (GSI) calculated. GSI from ELISA correlated with gonadal stage measured histologically. Monthly mean GSI of female oysters was highest during late spring to early summer (0·157–0·201) and lowest during early winter to early spring (0·002-0·000). Maximum GSI observed during the study was 0·422 for female oysters and 0·446 for male oysters. Female oysters produce 3·7–65·4 million eggs, with an average of 21·1 million during each spawning. A positive correlation was observed between the number of eggs produced and oyster size; the number of eggs in the gonad increased as oyster size (i.e. total dry wt) increased (r= 0·67); however, the relationship was non-linear. Large oysters contained proportionally fewer eggs. Prevalence of Perkinsus marinus parasitism was high, 90–100%, during the study, as was weighted incidence, 1·33 to 2·67, No statistically significant correlation was observed between infection intensity and the per cent weight of oyster eggs or egg number.     

Quantification of eggs and sperm in the Black-lip pearl oyster Pinctada margaritifera using an enzyme-linked immunosorbent assay (ELISA)

We have developed immunological probes to quantify eggs and sperm of the Black-lip pearl (BLP) oyster Pinctada margaritifera. The western blot assay revealed that the polyclonal antibodies developed in this study specifically recognized only egg and sperm proteins. These polyclonal antibodies also showed high sensitivities to the antigens, detecting 0.31–10 μg/ml of the egg or sperm proteins in an indirect enzyme-linked immunosorbent assay (ELISA). Accordingly, we used an indirect ELISA to quantify the eggs and sperm in BLP oysters collected in December 2009 from Weno Island in Micronesia and in May 2010 from Tahiti. The gonad somatic index (GSI), a ratio of gonad weight to somatic tissue weight, of the females collected from Weno Island ranged from 3.6 to 18.4 %, while the GSI of the females collected from Tahiti ranged from 5.6 to 12.6 %. Similarly, the GSI of the male BLP oysters from Weno Island ranged from 0.8 to 8.5 %, while the GSI of the males from Tahiti ranged from 4.8 to 7.5 %. These results lead to the conclusion that the immunological probes developed in this study can be successfully applied to quantify BLP oyster gonadal tissues and that these probes can be used in studies of BLP oyster reproductive ecology based on their sensitivity, rapidity, and affordability.     

Growth of the European flat oyster in the Mediterranean Sea (Murcia, SE Spain)

Growth of the natural European flat oyster (Ostrea edulis L.) spat from the Mar Menor (Murcia, Spain) was studied in the Mediterranean Sea over 18 months. The oysters were cultured in two types of containers, stackable plastic Galician trays and plastic mesh pots, deployed at a depth of 15 m hung from a long-line. Two size classes of spat were used, Class I spat (initially 53 mm in size and 19 g in weight) and class II spat (initially 31 mm, 4 g). The best results were observed in the class I oysters grown in the trays, which yielded a substantial biomass (24.77 kg m^–2), the survival rate was 69% and commercial size (60 mm) was attained by all the oysters within 9 months of the start of the experiment. The observed growth of the class II oysters was poor, attaining a mean of not more than 50 mm and 18 g; the survival rate ranged between 25–74% and only between 8–13% of the class II spat attained commercial size.     

Sexing accuracy and indicators of maturation status in captive Murray cod Maccullochella peelii peelii using non-invasive ultrasonic imagery

Macroscopic- and histological-based assessments of gonad condition were compared with ultrasound images to determine the feasibility of this technology as a non-invasive diagnostic tool for identifying sex and assessing maturation status of Murray cod. Four age-classes (1+, 2+, 3+ and 6+ years), were sub-sampled at monthly intervals throughout their annual reproductive cycle and scanned with a 5 MHz linear transducer. An interpretation of sex was made from the resulting images and maximum cross-sectional gonad diameter and area were recorded. Fish were subsequently dissected to confirm gender, and the weights and maturation status of gonads determined and then compared with their respective image profile. Ovaries of females were usually a distinctive feature in ultrasound images, being particularly obvious in older and/or more developed fish. In contrast, the identification of male testis was more problematic. Nonetheless, identifying sex from ultrasound images was consistently achieved by recording the presence/absence of a female ovary (96% total sexing accuracy). Maximum cross-sectional ovary diameter and area were highly correlated with gonad weight (r² = 0.90 and 0.89, respectively) suggesting that indices of maturation status, comparable to the gonadosomatic index (GSI), can be obtained non-destructively from ultrasound scans of females. A less distinct relationship occurred between these dimensions and weight of testes (r² = 0.41). Significant increases (P < 0.05) in mean gonad index (GI, calculated from gonad diameter) occurred for most gonad development stages. However, differences in mean GI between maturation stages were confounded by phenotypic variability, indicating that GI may be limited to population level studies. Nevertheless, ultrasound images of ovaries at each development stage were visually distinctive and enabled qualitative evaluations of maturity, thereby complementing quantitative GI assessments. Repeated serial-monitoring of the same population using ultrasound appears to have great potential for tracking maturation-induced changes in broodfish.     

Early growth and reproduction of hatchery-produced Pacific oyster Crassostrea gigas in Gamakman Bay off the southern coast of Korea

We investigated the early growth and reproduction of hatchery-produced Pacific oysters Crassostrea gigas raised in a suspended long-line facility in Gamakman Bay, off the south coast of Korea. In October 2009, 4?months after transplanting, shell length had increased from 27.4 (July) to 82.5?mm (October), and tissue weight had increased from 0.2 to 5.2?g, indicating that the oyster had reached a marketable size in this month. Histological studies indicated rapid gonad maturation, and the oysters spawned during August and October, with a peak in September. An enzyme-linked immunosorbent assay used to quantify egg biomass revealed that the oysters produced a relatively small quantity of eggs, ranging from 5.1?% (August) to 8.8?% (September) of their body weight. The low total carbohydrate reserve in the tissue recorded in August and September coincided with intense energy utilization due to spawning, while the protein maximum in September matched peak egg mass. Our results suggest that hatchery-produced seed could supply a portion of the spat required in Gamakman Bay as well as in other oyster culture grounds of Korea, where the oyster industry is facing a shortage in the supply of natural spat.     

Family by environment interactions in shell size of 43-day old silver-lip pearl oyster (Pinctada maxima), five families reared under different nursery conditions

To understand the influence the environment and associated genotype by environment interactions will exert on future silver-lip pearl oyster (Pinctada maxima) selective breeding programs, this study assessed the relative performance in four shell growth traits of spat from five full-sib families, when spat were communally reared at different salinities (29, 34 and 40 ppt), food availability (high, medium and low), food quality (high, medium and low), and in a hatchery vs. ocean environment for 43 days.Rearing environment was found to influence growth expression, with significant differences evident when oysters were grown at different salinities in the ocean instead of hatchery, or when fed algae of differing nutritional quality. As indicated by MANOVA, family comparative growth performances were also altered when the environment changed, with significant environment by family interactions apparent in the food quality, food availability and hatchery vs. ocean rearing treatments. Changes in salinity, however, did not affect relevant family performances.These results indicate that growth and relative family performance in P. maxima may change dependent on local environmental conditions and that genotype by environment effects may need to be considered in breeding programs for this species.     

Tetraploid induction in tropical oysters,Crassostrea belcheri (Sowerby) and Crassostrea iredalei (Faustino)

Tetraploid induction has been conducted on temperate oysters but not on tropical oysters. In this study, different heat shocks (32, 35 and 38°C) and cold shocks (1, 4 and 7°C) were used to induce tetraploidy in two tropical oyster species, Crassostrea belcheri and Crassostrea iredalei, through meiosis I inhibition. Temperature shocks were applied on the newly fertilized eggs at 8–10 min post fertilization and terminated when second polar bodies began to form in the control eggs. The ploidy of the larvae and spat was determined via direct chromosome count. The percentage of larval survival until Day 20 was low (between 0.4% and 42.9%) for both temperature shocks and oyster species. No surviving larva was recorded for induction at 1, 4 and 38°C. Tetraploid spat was only recorded in C. iredalei but the percentage is low through heat shock induction of 32 and 35°C. This study shows that the tetraploid induction success rate was slightly higher in C. iredalei compared to C. belcheri. No surviving tetraploid spat were recorded for both oyster species through the cold shock method. This study shows that heat shock can be used to inhibit meiosis for the production of tetraploids but more experiments need to be conducted to determine the optimum temperature when dealing with tropical oysters.     

中华花鳅的繁殖生物学特性研究

探究中华花鳅(Cobitis sinensis)的繁殖生物学特性,可为其人工增养殖和资源保护利用提供科学依据。2017年11月至2018年5月,在贵州境内舞阳河施秉段采集中华花鳅样本381尾,以鳞片作为年龄鉴定材料,将样本解剖后进行性别鉴定和性腺分期。结果表明,舞阳河中华花鳅体长(L)为58.9~100.2 mm,体重(W)为2.0~13.7 g,年龄在1~4龄。381尾样本中,性别不可辨样本有23尾,雄248尾,雌110尾,性腺发育至IV期最多。经肉眼观察发现,中华花鳅的精巢和卵巢均可分为6期,有70尾雌鱼性腺发育达到IV期,其绝对繁殖力(F)为542~2 865粒,体长相对繁殖力(FL)为6.6~28.6粒/mm,体重相对繁殖力(FW)为99.8~490.7粒/g;绝对繁殖力(F)、体长相对繁殖力(FL)均与体长(L)、体重(W)、净体重(W0)、性腺重(W1)、成熟系数(GSI)之间具有极显著相关性(P<0.01),体重相对繁殖力(FW)与性腺重(W1)、成熟系数(GSI)之间具有极显著相关性(P<0.01)。实地调查发现,中华花鳅的产卵场主要在砂质底质、水质清新、水流较缓、水草丰富的浅水区,繁殖期为5-8月,繁殖高峰期为5月中旬至6月上旬。研究显示,中华花鳅的繁殖力较低,繁殖高峰期时间较短,低龄化严重,亟需对其进行资源保护和系统研究。     

Parasites, pathological conditions and mortality in QX-resistant and wild-caught Sydney rock oysters, Saccostrea glomerata

Differences in the survival of QX-resistant fifth generation (QXR₅) and wild-caught (Wild-Caught) Sydney rock oysters were assessed over the QX-disease risk period in the Pimpama River, SE Queensland. Cumulative mortality of Wild-Caught oysters (31.7%) was significantly greater than QXR₅ oysters (0.0%) over the 118 days of the experiment. PCR and histological results showed that Wild-Caught oyster did not die from QX disease. Histology revealed oysters were infected with disseminating hemocytic neoplasia, a Steinhausia-like infection, a Rickettsia-like organism infecting epithelial cells of the gill, digenean flukes encysted in the gonadal tissue and a gill response to an unknown toxin. The cause of mortality is attributed to disseminating hemocytic neoplasia.     

Suspended Culture of Doughboy Scallops Mimachlamys asperrima (Lamarck)

To evaluate the effects of suspended culture on the wet meat weights, reproductive condition and fecundity of doughboy scallops Mimachlamys asperrima , scallops held either in suspended cages or collected from wild stocks on the sea bed were compared over a year. Changes in gonosomatic index (GSI) and macroscopic staging of gonads were similar in both populations; however, after one month, soft body, muscle and gonad weights were heavier in suspended scallops and, with the exception of gonad weight in early summer, remained heavier throughout the year. Fecundity following serotonin-induced spawning of suspended Mimachlamys asperrima was on average almost twice that of similar sized scallops grown on the sea bed (1.58 and 0.84 million eggs/female, respectively). Suspended scallops surpassed all previous maxima for tissue weights, GSI and fecundity recorded for wild Jervis Bay Mimachlamys asperrima . This indicated that farmers could obtain greater meat yields from suspended rather than reseeded or wild Mimachlamys asperrima and that broodstock to be used in hatcheries may be best obtained from suspended culture.     

Influence of gonadal development on metal accumulation in the Portuguese oyster,Crassostrea angulata,in subtropical areas

Metal enrichment makes oyster considered to be an appropriate food source for trace elements, but excessive accumulation can threaten human health. Reproduction is an important biotic factor that affects the metal accumulation in oysters. However, the influence of continuous gonad development model on metal accumulation in some subtropical oysters is still unclear. The present study addresses the concentrations of trace metals and glycogen associated with the gonadal and somatic tissues throughout the reproductive cycle of diploid and triploid Crassostrea angulata cultured in Fujian, China. Cu, Zn and Pb contents of gonads were negatively correlated (p < 0.05) with gonadosomatic index (GSI). The correlation between Cd content and GSI was not significant. There was a positive correlation (p < 0.05) between GSI and Mn content in female gonads while the correlation was negative (p < 0.05) in males, indicating Mn plays an important role in gametogenesis of females. The effects of gonadal maturity and gender on metal accumulation in somatic tissue were not significant (p > 0.05). The concentrations of all metals in sterile triploids were not significantly different from those of the somatic tissue of diploids. Gonadal glycogen content showed a negative correlation with the GSI. However, somatic glycogen content was not affected by gonadal development, indicating that the energy consumed during gametogenesis comes mainly from recently ingested food rather than from energy stored in somatic tissue. The effects of the gonadal development on the metal accumulation in oysters varied according to the metal and tissue type.     

Seasonal changes in hepatosomatic index,gonadosomatic index and plasma estradiol‐17β level in captively reared female rabbit fish (Siganus guttatus)

The seasonal changes in hepatosomatic index (HSI), gonadosomatic index (GSI) and plasma estradiol‐17β (E₂) level in female rabbit fish (Siganus guttatus) were investigated. The relationship between plasma E₂ levels with these indices and ovarian growth was also evaluated. Each month, at least 10 female broodfish were sacrificed to collect liver, ovary and blood for HSI, GSI and plasma E₂, respectively. GSI and HSI were calculated as percentage (%) of relative weight of gonad and liver to total body weight, respectively. Plasma E₂ level was measured using enzyme‐linked immunosorbent assay method (ELISA). Ovaries were cut and stained for histological observation. The results included seasonal changes in plasma E₂ levels, stages of ovarian development, GSI and HSI. The highest level of E₂ was observed in June (1,445.62 pg/ml) and during vitellogenesis (2,305 pg/ml). GSI and HSI values significant fluctuated monthly. The highest HSI and GSI were 1.72% in May and 3.58% in June, respectively. The pattern of plasma E₂ levels showed a relationship with GSI and different stages of ovarian development. HSI was associated with ovarian stages. During vitellogenesis, the highest value (1.9%) of HSI was observed. Histological sections showed that rabbit fish is a multiple spawner. These results contribute to further understanding of female rabbit fish reproductive biology in captivity. Important reproductive parameters such as HSI, GSI and E₂ can be used to indicate maturation status of this fish species.     

Reproductive biology of the commercial sea cucumber <Emphasis Type="Italic">Holothuria spinifera</Emphasis> (Echinodermata: Holothuroidea) from Tuticorin,Tamil Nadu,India

The annual reproductive cycle of the commercial sea cucumber Holothuria spinifera was studied in Tuticorin, Tamil Nadu, India, from September 2000 to October 2001, by macroscopic and microscopic examination of gonad tubule, gonad index and histology of gametogenic stages, to determine the spawning pattern. The gonad consists of long tubules with uniform development. It does not confirm the progressive tubule recruitment model described for other holothurians. The maximum percentage of mature animals, gonad and fecundity indices, tubule length and diameter, with the observations on gonad histology, ascertained that H. spinifera had the peak gametogenic activity during September and October 2001 followed by a prolonged spawning period from November 2000–March 2001.     

Off-Season Spawning of Eurasian Perch Perca Fluviatilis

This study was designed to determine the effects of the selection for body weight and of two final warming durations during an out-of-season reproductive cycle on gonad recrudescence, plasma steroid levels and spawning in Eurasian perch Perca fluviatilis males and females. Two different bodyweight classes (200–250 and 350–400 g) and two different warming periods (1 or 2 months) were studied.At the end of the chilling period, a significant effect of the selection for body weight was observed on gonad development whereas fish were the same age. According to higher levels in testosterone (12 ng mL^–1), a better development was recorded for smaller breeders (P < 0.05). A short water warming period up to 14°C (1 month v.s. 2 months) allowed higher rates of females with completed gonadogenesis (33–38% v.s. 7–8%) and spawning (31–33% v.s. 0%) (P < 0.05). Spontaneous out-of-season spawning (17 ribbons) and larvae were obtained. Relative fecundity was about 100 eggs g^–1 of female body weight. Five ribbons were partly fertilized with a rate ranging from 5 to 90%. These studies have shown that delayed spawning in Eurasian perch can be obtained by a temperature manipulation, but results are not sufficient because of low stimulation rate of females, low spawning rates and high heteroge-neity of fertilization rates. The effects of other environmental factors like photoperiod should be tested in further experiment to improve the protocol inducing out-of-season spawning production.     

Reproductive condition of the pearl oyster, Pinctada imbricata, Röding, in Port Stephens, New South Wales, Australia

In an investigation of the potential for pearl production in New South Wales (NSW), Australia, changes in the physical and reproductive condition of the pearl oyster, Pinctada imbricata, were monitored for over 2 years. Using wild oysters gathered from close to the southern extent of the species' range in Port Stephens, NSW, a series of macroscopic and histological observations were made. Reproductive activity in P. imbricata was greatest from late spring to early autumn with oysters in poor reproductive condition during winter. Peaks in reproductive indices occurred in November 1998, March 1999, December 1999 and April 2000. Four indices of physiological condition were used: shell growth, byssal attachment, mantle thickness and mucoprotien layer. With the exception of the thickness and extent of the mucoprotein layer, these indices either showed little variation or the variation that occurred was not related to seasonal or reproductive changes. Changes in the mucoprotein layer were correlated with water temperature and suggest that this layer is metabolized during periods of high demand such as during gonadogenesis. Collectors deployed at two sites in Port Stephens demonstrated that spatfall occurs largely in the months of December and January following the spring and early summer peaks in reproductive activity (November 1998 and December 1999). Spatfall was not observed following the autumnal peaks (March 1999 and April 2000) in reproductive activity. Overall, reproductive patterns in P. imbricata are poorly suited to spat supply in Port Stephens. Farmers require spat in early spring (September) to allow maximum use of the ‘growing’ season (September–May). Reproductively capable oysters are not available from the wild until September and quantity of natural spatfall is too variable and occurs too late in the season (December–January). As a result, oysters are being conditioned in the hatchery in July, spawned in August and spat are supplied to farmers in mid September.     

Intra‐annual changes in reproductive indices of male and female Himalayan snow trout,Schizothorax richardsonii (Gray, 1832)

Periodic changes in reproductive hormone levels, gonadal histology and gonadosomatic index (GSI) of snow trout, Schizothorax richardsonii, were examined to ascertain annual cycle of gonadal development and reproductive status in their natural habitat. In females, there were coherent changes in plasma 17β‐oestradiol and vitellogenin along with GSI, oocyte maturation and vitellogenic progression, collectively indicating two distinct maturation peaks during the months of September and February. Coinciding with this, in males, plasma 11‐keto testosterone was also noticeably higher during September and February, with highest GSI values in September. However, plasma 17α, 20β‐dihydroxyprogesterone levels in males were found to be persistently high from September to February. This observation suggests the potential presence of matured oozing males over a longer period, unlike in females. Overall, the close association between reproductive hormone levels, GSI and gonadal maturation stages in males and females (particularly, the presence of postovulatory follicle complexes) with apparent natural synchronization clearly indicates that S. richardsonii breeds twice in a year, possibly during late September to early November and late February to early April in the coldwater riverine habitats of the Indian Himalayan region.     

Relationships among environment, spawning and settlement of Queen scallop in the Ría de Arosa (Galicia, NW Spain)

This study examines the relationship of the reproductive and reserve storage cycles and spat settlement of the Queen scallop (Aequipecten opercularis) to environmental factors. The possibility of farming this species from spat settled on collectors is considered. The gonad condition index shows maximum values between mid-May and early August. Another peak occurs in mid-November. In spring and summer, the energy needed for gametogenesis is derived from food, although it is possible that direct transfer may occur from the digestive gland to the gonad. The glycogen and lipid content of the reserve organs reaches maximum values between late summer and early autumn. The energy requirements related to gametogenesis, which begins in autumn, involve the consumption of accumulated reserves with the resulting reduction in the condition indices. There is a positive correlation between temperature and the condition indices of the digestive gland, the adductor muscle and their lipid and glycogen content, respectively. Two settlement periods were recorded, one in winter, which could not be associated with the spawning of the experimental animals, and another in summer, which coincided with the spawning that took place in spring. The reduced gonad indices recorded at the end of the summer and in winter did not lead to settlement.     

长江中游贝氏(鳖)繁殖生物学研究

2016年4-12月在监利长江何王庙故道采集591尾样本,对长江中游贝氏(鳖)(Hemiculter bleekeri)性比、初次性成熟大小、繁殖季节、产卵类型、产卵群体组成及繁殖力等繁殖生物学指标进行了分析.结果显示:贝氏(鳖)总体雌雄性比1.44:1,与1:1存在极显著性差异;繁殖期间雌雄比2.5:1,非繁殖季节雌雄比为0.91:1.繁殖群体年龄结构为1-3龄,以1龄补充群体为主,属于第Ⅱ类型.初次性成熟体长:SL♀50=8.08 cm, SL♂50=8.47 cm,最小观测性成熟个体分别为:雌性7.5 cm,体重6.65 g, 1龄;雄性8.2 cm,体重8.13 g, 1龄.根据成熟系数变化和性腺发育期观察,推测其繁殖季节为4-6月,可能延至7月份.贝氏(鳖)绝对繁殖力为1703~9601粒/尾,平均为(5876.6 ± 1837.8) 粒/尾,与体重、性腺重均呈线性关系,与体长呈幂函数关系.相对繁殖力(FW)为204.7~406.5粒/g,平均(322.3 ± 49.7) 粒/g,与成熟系数呈幂函数相关;相对繁殖力(FL)为212.0~827.6粒/cm,平均(552.4 ± 136.1) 粒/cm,与体重幂函数相关.     

Gametogenesis, reproductive investment, and spawning behavior of the Pacific giant oyster Crassostrea gigas: evidence of an environment-dependent strategy

The progress of gametogenesis was studied in oysters Crassostrea gigas having the same origin (Tremblade), but cultured during 1 year in two distinctive French marine areas, the Baie des Veys and Marennes-Oléron. We assessed seasonal changes in the reproduction cycle on the basis of stereological techniques to estimate reproductive investment and measurement of gonad evolution area by quantitative histology. From a qualitative point of view, both oyster groups presented typical reproductive stages, but showed differential timing, in particular during the sequence of spawning and duration of the re-absorption stage. Oysters in Baie des Veys had a single partial spawning in August and a re-absorption stage that extended until winter. Oysters in Merennes-Oléron had a partial spawning in July and massive release of gametes during August. Spawnings in both the areas were related to maximum temperature (19°C). The quantitative analysis showed, on an annual basis, a higher reproductive investment by oysters from Baie des Veys, 86% against 53% in the other group. Larger gonads, higher gamete production, and more intensive spawning were the characteristics of oysters in Baie des Veys. Recently, the reproduction pattern and investment has been related with summer mortalities; therefore, a quantitative understanding of reproductive processes becomes necessary for C. gigas. Environmental conditions at each site may explain differences in the progress and intensity of gametogenesis. While temperature regulated the time and speed of gametogenesis, results suggest that the intensity was influenced by the quantity of available food but may need further research. However, nutrient recycling from unreleased gametes in the gonads of oysters from Baie des Vey is a factor to be considered in the results of this study.     

Gonadosomatic index and gonad histopathology of Gobius niger (Gobiidea, Teleost) from Bizerta lagoon (Tunisia): Evidence of reproduction disturbance

To evaluate the effect of anthropogenic pollution on fish reproduction in Bizerta lagoon, one of the largest Tunisian lagoons subjected to various anthropogenic and industrial pressures, the gonadosomatic index (GSI) and gonad histology of mature black goby (Gobius niger) were studied. Fish were collected during the breeding seasons from six stations differently impacted and in one reference station located at the seaward entrance of Ghar el Melh (GH) lagoon. In both sexes, we detected the presence of an overall weaker GSI in Bizerta lagoon than in control fish. These GSI alterations were accompanied by histopathological changes. In females, no significant differences in oocyte size were found across all the sample sites. However, a higher incidence of oocyte lesions (atretic oocytes, cytoplasmic retraction and karyoplasmic cluping) was found in stations located near agricultural activity zones. Histological observation of the testes revealed, as well, abnormalities in Bizerta lagoon fish: germ cells syncytium, vacuolated germ cells, melano-macrophage center and increase of gaps in the interstitium between lobules were detected with a high prevalence in stations influenced by industrial and/or urban activities. In addition, association between gonad fish histopathology and the presence of organic pollutants (DTT, PAHs and organotins) in sediment was noted. These results suggest that the chemical pollution in this lagoon may have adversely affected the gonad development of fish. In conclusion, this study is the first histological evidence of reproduction disturbance related to pollution pressure in a fish species inhabiting Bizerta lagoon. The high levels of gonad histopathology recorded raise concerns about the long-term health of fish populations in this lagoon.