Coral reef ecosystems of Reserva Natural de La Parguera (Puerto Rico): spatial and temporal patterns in fish and benthic communities (2001-2007)

Since 1999, NOAA’s Center for Coastal Monitoring and Assessment, Biogeography Branch (CCMA-BB) has been working with federal and territorial partners to characterize monitor and assess the status of the marine environment in southwestern Puerto Rico. This effort is part of the broader NOAA Coral Reef Conservation Program’s (CRCP) National Coral Reef Ecosystem Monitoring Program (NCREMP). With support from CRCP’s NCREMP, CCMA conducts the “Caribbean Coral Reef Ecosystem Monitoring project” (CREM) with goals to: (1) spatially characterize and monitor the distribution, abundance and size of marine fauna associated with shallow water coral reef seascapes (mosaics of coral reefs, seagrasses, sand and mangroves); (2) relate this information to in situ fine-scale habitat data and the spatial distribution and diversity of habitat types using benthic habitat maps; (3) use this information to establish the knowledge base necessary for enacting management decisions in a spatial setting; (4) establish the efficacy of those management decisions; and (5) develop data collection and data management protocols. The monitoring effort of the La Parguera region in southwestern Puerto Rico was conducted through partnerships with the University of Puerto Rico (UPR) and the Puerto Rico Department of Natural and Environmental Resources (DNER). Project funding was primarily provided by NOAA CRCP and CCMA. In recent decades, scientific and non-scientific observations have indicated that the structure and function of the coral reef ecosystem in the La Parguera region have been adversely impacted by a wide range of environmental stressors. The major stressors have included the mass Diadema die off in the early 1980s, a suite of hurricanes, overfishing, mass mortality of Acropora corals due to disease and several coral bleaching events, with the most severe mass bleaching episode in 2005. The area is also an important recreational resource supporting boating, snorkeling, diving and other water based activities. With so many potential threats to the marine ecosystem several activities are underway or have been implemented to manage the marine resources. These efforts have been supported by the CREM project by identifying marine fauna and their spatial distributions and temporal dynamics. This provides ecologically meaningful data to assess ecosystem condition, support decision making in spatial planning (including the evaluation of efficacy of current management strategies) and determine future information needs. The ultimate goal of the work is to better understand the coral reef ecosystems and to provide information toward protecting and enhancing coral reef ecosystems for the benefit of the system itself and to sustain the many goods and services that it offers society. This Technical Memorandum contains analysis of the first seven years of fish survey data (2001-2007) and associated characterization of the benthos. The primary objectives were to quantify changes in fish species and assemblage diversity, abundance, biomass and size structure and to provide spatially explicit information on the distribution of key species or groups of species and to compare community structure across the seascape including fringing mangroves, inner, middle, and outer reef areas, and open ocean shelf bank areas.

A sudden collapse in distribution of Pacific sardine (Sardinops sagax) off southwestern Australia enables an objective re-assessment of biomass estimates

Fishery-independent estimates of spawning biomass (BSP) of the Pacific sardine (Sardinops sagax) on the south and lower west coasts of Western Australia (WA) were obtained periodically between 1991 and 1999 by using the daily egg production method (DEPM). Ichthyoplankton data collected during these surveys, specifically the presence or absence of S. sagax eggs, were used to investigate trends in the spawning area of S. sagax within each of four regions. The expectation was that trends in BSP and spawning area were positively related. With the DEPM model, estimates of BSP will change proportionally with spawning area if all other variables remain constant. The proportion of positive stations (PPS), i.e., stations with nonzero egg counts — an objective estimator of spawning area — was high for all south coast regions during the early 1990s (a period when the estimated BSP was also high) and then decreased after the mid-1990s. There was a decrease in PPS from the mid-1990s to 1999. The particularly low estimates in 1999 followed a severe epidemic mass mortality of S. sagax throughout their range across southern Australia. Deviations from the expected relationship between BSP and PPS were used to identify uncertainty around estimates of BSP. Because estimation of spawning area is subject to less sampling bias than estimation of BSP, the deviation in the relation between the two provides an objective basis for adjusting some estimates of the latter. Such an approach is particularly useful for fisheries management purposes when sampling problems are suspected to be present. The analysis of PPS undertaken from the same set of samples from which the DEPM estimate is derived will help provide information for stock assessments and for the management of purse-seine fisheries.

Environmental parameters and incidence of white spot disease in Penaeus monodon (Fab.) farming

An investigation was carried out to monitor management practices and to find out whether there is any relationship with occurrence of deadly white spot disease and environmental parameters. Three semi-intensive and a improved traditional shrimp farms were selected in which mass mortality of shrimp (Penaeus monodon) by white spot disease occurred previously. The farms were situated at two different geographical locations. Two ponds from each farm at random were selected for the study. Out of eight investigated ponds, 6 ponds in three farms were affected by the disease during investigation period. The non-affected ponds had relatively lower stocking density, lightly different management practice and were located at different geographical area. There was no significant variation in water quality parameters among the affected and non-affected ponds. No significant variations were recorded in pond preparation, source of Post Larvae (PL), water and feed management among the affected and non-affected ponds. The observation indicated that pond micro-organisms in a farm may not the only cause of the disease but some external factors also might be responsible for the outbreak of this disease.

A comparative study on the embryonic development of gynogen, triploid, haploid and normal diploid embryos of stinging catfish, Heteropneustes fossilis

UV irradiation and cold shock were applied on the eggs of stinging catfish, Heteropneustes fossilis, to produce haploid,. gynogen and triploid embryos. A comparative account of the various features· of embryonic development in chromosomally manipulated groups viz. haploid, gynogen and triploid and non-manipulated normal diploid group of H fossilis has been discussed. A slow development and delayed hatching were observed in gynogen and triploid embryos compared to those in normal diploid (control) groups. Mass mortality was observed in all chromosomally manipulated groups particularly during the gastrulation stage. The hatchlings of the gynogen, triploid and normal diploid were similar in overall appearance.

Organic pollution in culture water resulting from excess feed and metabolite buildup

Penaeus monodon postlarvae were subjected to increasing feed concentrations and their growth and survival rates were recorded. Measurements were made of dissolved organic matter, and ammonia and nitrite-nitrogen concentrations. Survival was highest at the lowest feeding level and decreased as feed concentration increased. It is concluded that although organic matter enriches the food supply for P. monodon postlarvae, at higher concentration levels it can pollute the culture water, which in turn leads to mass mortality of the postlarvae. Secondly, the survival rate of P. monodon postlarvae is directly related to dissolved organic matter concentration, oxygen tension, and ammonia-nitrogen concentrations in the culture water. Even at sublethal levels these adverse environmental conditions decrease the survival rate.

Observations on the artificial fertilization of eggs and larval rearing of the grey mullet, Mugil cephalus L.

The results are presented of attempts to artificially fertilize Mugil cephalus eggs in the Philippines. Embryonic development is outlined and rearing of the larvae described. Mass mortality occurred during week 3 of rearing.

Impact of mangrove conversion

The article presents the impact of mangrove conversion on fisheries and on coastal areas. The mangrove areas which serve as nursery grounds for important species of fish and crustaceans are also rich feeding ground for many species from various trophic levels. Thus, the destruction of mangroves could affect the availability of fry and broodstock and, consequently, aquaculture production and fisheries. While in coastal areas, the destruction of mangroves increased the risk of coastal erosion from storm surges and winds, accelerates the erosion of riverbanks, exposes acid sulfate soils, leading to poor production and mass mortality of stocks, and affects the freshwater supply through salt intrusion upstream among others.

Construction of AFLP-based genetic linkage map for Zhikong scallop, Chlamys farreri Jones et Preston and mapping of sex-linked markers

Zhikong scallop (Chlamys farreri) is an economically important aquaculture species in China; however, frequent mass mortality seriously affects the development of its industry. Genetic linkage map is useful for genetic improvement and selective breeding of C. farreri. Linkage maps were constructed using an intraspecific F-1 cross and amplified fragment length polymorphism (AFLP) markers. Thirty-two selected AFLP primer combinations produced 545 AFLP markers that were polymorphic in either of the parents and segregated in the progeny. Of these segregating markers, 166 were mapped to 19 linkage groups of the female framework map, covering a total of 1503.9 cM, with an average marker spacing of 10.2 cM; and 197 markers were assigned to 20 linkage groups of the male map, covering a total of 1630.7 cM, with 9.2 cM per marker. A sex-linked marker was mapped on the female map with zero recombination and a LOD of 27.3. The genetic length of C farreri genome was estimated as 1889.0 cM for the female and 1995.9 cM for the male. The coverage of the framework map was calculated as 79.6% for the female and 81.7% for the male. When the triplets and doublets were considered, the observed length of the map was calculated as 1610.2 cM with coverage of 85.2% for the female, and 1880.5 cM with coverage of 94.2% for the male. The genetic maps presented here will serve as a basis for the construction of a high-resolution genetic map and mapping of economically important genes. (C) 2004 Published by Elsevier B.V.

Physiological and immune responses of zhikong scallop Chlamys farreri to the acute viral necrobiotic virus infection

The Zhikong Scallop, Chlamys farreri, is one of the most Important bivalve mollusks cultured in northern China However, mass mortality of the cultured C farreri has posed a serious threat to the maricultural Industry in recent years. Acute Viral Necrobiotic Virus (AVNV) is believed as an important etiological agent causing the scallop mass mortalities To understand the mechanism behind the AVNV associated scallop disease and mortality, we assessed the physiological and immune responses of C farreri to the virus infection using oxygen consumption rate, ammonium-nitrogen excretion rate, hemocyte copper, zinc superoxide dismutase gene expression, and plasma superoxide dismutase activity and alkaline phosphatase activity as indicators Scallops challenged by AVNV at 25 C developed typical disease signs 2 days after virus injection Before the disease manifested, scallop oxygen consumption and NH4+-N excretion rates rose and then fell back. Real-time PCR revealed that the hemocyte cytosol Cu, Zn SOD gene expression was upregulated followed by recovery The plasma SOD activity, however, augmented consistently following virus injection Moreover, plasma AKP activity first lowered and then elevated gradually to the highest level at 24 h post virus injection Scallops challenged by AVNV at 17 degrees C neither developed notable disease nor showed obvious responses that could be associated with the virus infection. While the results suggested a correlation between the elevated seawater temperature and the AVNV infection associated C farreri mortalities, they also indicated that the viral infection provoked multiple physiological and immune responses in the host scallops (C) 2010 Elsevier Ltd All rights reserved

栉孔扇贝大规模死亡病原学研究

用平板画线法从患病栉孔扇贝（Chlamys farreri）体内分离到了一种原核生物（简称QDP）。QDP可以在改进的液体培养基MEM（含2.2%NaCl，5%小牛血清）和脑心浸液（含2.2% NaCl）中生长；菌落在显微镜下（150×）为无色、透明的小点状；革兰氏染色阴性；菌体为圆形或近似圆形。QDP在发育过程中有两种状态，一种为未成熟阶段，直径小于100nm；另一种为成熟阶段，直径变化很大，最小约60nm，最大可达4µm以上。较小的个体有拟核、核糖体和新月状的空泡，未见细胞壁；较大的个体有细胞壁，胞内大部分被空泡充满，未见拟核和核糖体。栉孔扇贝组织超簿切片电镜观查证实QDP的存在。QDP的密度随着生长发育时间的不同而有所变化，繁殖高峰期密度较大。 建立了密度梯度离心结合滤膜过滤分离技术，优化人工培养条件。最适生长温度为23℃，最适生长pH值为7.4，最适生长盐度相当于细胞培养液所需的盐浓度（0.85%NaCl）。 提取的QDP核酸能被RNase A 降解，且没有检测到DNA。以PCR、RT-PCR扩增其16SrRNA基因序列片段，PCR反应没有扩增出扩增子，而RT-PCR则扩增出了16S rRNA基因序列片段，经测定其序列全长度为1430bp，经与GENEBANK中的16S rRNA片段比较分析，与6种不同科的微生物的同源率最高的为95%-95.47%。 采用温度梯度和病原浓度梯度回归感染实验方法，较为系统地研究了QDP的致病性。研究结果表明：QDP对栉孔扇贝有强烈的致病作用，高温（23℃以上）是其致病的必要条件，证实DQP是栉孔扇贝大规模死亡的病原体之一。

Survival, growth and immune activity of scallop Chlamys farreri cultured at different depths in Haizhou Bay (Yellow Sea, China) during hot season

Survival, growth and immune response of the scallop, Chlamys farreri, cultured in lantern nets at five different depths (2, 5, 10, 15, and 20 m below the sea surface) were studied in Haizhou Bay during the hot season (summer and autumn) of 2007. Survival and growth rates were quantified bimonthly. Immune activities in hemolymph (superoxide dismutase (SOD) and acid phosphatase (ACP)) were measured to evaluate the health of scallops at the end of the study. Environmental parameters at the five depths were also monitored during the experiment. Mortalities mainly occurred during summer. Survival of scallops suspended at 15 m (78.0%) and 20 m (86.7%) was significantly higher than at 2 m (62.9%), 5 m (60.8%) or 10 m (66.8%) at the end of the study. Mean shell height grew significantly faster at 10 m (205.0 mu m/d) and 20 m (236.9 mu m/d) than at 2, 5 or 15 m in summer (July 9 to September 1); however, shell growth rate at 20 m was significantly lower than at the other four depths in autumn (September 2 to November 6). In contrast to summer, scallops at 5 m grew faster (262.9 mu m/d) during autumn. The growth of soft tissue at different depths showed a similar trend to the shell. Growth rates of shell height and soft tissue were faster in autumn than in summer, with the exception of shell height at 20 m. SOD activity of scallops increased with depth, and ACP activity was significantly higher at 15 and 20 m than at other depths, which suggests that scallops were healthier near the bottom. Factors explaining the depth-related mortality and growth of scallops are also discussed. We conclude that the mass mortality of scallop, C. farreri, during summer can be prevented by moving the culture area to deeper water and yield can be maximized by suspending the scallops in deep water during summer and then transferring them to shallow water in autumn.

Haematological responses of Penaeus monodon to environmental alterations and pathogenic invasion

This thesis Entitled Haematological responses of penaeus monodon to environmental alterations and pathogenic invasion. Thesis concluded from the present study that stress is accompanied by alterations in haemolymph metabolic variables and immune responses that influences the susceptibility of P. monodon to infection. Acute salinity variations were proved to be a stress condition that enhances the susceptibility of P. monodon to V. harveyi and WSSV infection. Ambient Cu at 0.1 mg 1" and ambient Zn at 1.0 mg 1" proved immunostimulatory in increasing the immunocompetence of P. monodon to WSSV infection and higher concentrations of Cu and Zn proved immunosuppressive. Haemolymph total protein, total carbohydrates and total lipids showed the highest relation with immune responses. THC, PO, ACP and ALP that greatly correlated with the survival rate proposed as reliable biomarkers of health in P. monodon. The study highlights the need for proper management practices and regular health monitoring to be adopted to avoid mass mortality in shrimp culture ponds.

Vibrio harveyi phages: Isolation, characterization and evaluation of their potential as phage therapeutics on Vibrio harveyi in shrimp hatcheries

Aquaculture is a global industry providing food and employment thereby contributing to the economy. For the sustenance of aquaculture, disease management is a major requirement. Among the bacterial pathogens Vibrio harveyi remains to be the major one especially in shrimp culture systems. Rapid and mass mortality of shrimp larvae due to Vibrio harveyi infection is well known, and the pathogen causes serious economic losses in grow out systems as well. It suggests that a well defined management strategy has to be built up to protect the crop from Vibrio harveyi infection in aquaculture systems. Antibiotics have been the choice for quite some times which led to residues in meat and development of multidrug resistant bacteria which invited ban on their application. In this context several alternate options have been thought off such as probiotics, immunostimulants and vaccines. Phage therapy is yet another option. Phages being natural parasites of bacteria and are abundant in aquatic environments their application to control bacterial pathogens in aquaculture has commendable potential in lieu of antibiotics. For that matter the therapeutic effect of phages has been proven in several antibiotic resistant pathogens inclusive of Vibrio harveyi.

Isolation of a pathogenic strain of Vibrio alginolyticus from necrotic larvae of Macrobrachium rosenbergii (de Man)

Giant freshwater prawn, Macrobrachium rosenbergii (de Man), is an important commercial species with considerable export value, ideal for cultivation under low saline conditions and in freshwater zones (Kurup 1994). However, despite more than a decade of research on its larval production systems, vibriosis still hampers seed production resulting in high mortality rates. Among the different species of vibrios, Vibrio alginolyticus has been isolated frequently from diseased shrimp as the aetiological agent of vibriosis and has been described as a principal pathogen of both penaeids and nonpenaeids (Lightner 1988; Baticados, Cruz-Lacierda, de la Cruz, Duremdez-Fernandez, Gacutan, Lavilla- Pitogo & Lio-Po 1990; Mohney, Lightner & Bell 1994; Lee, Yu, Chen, Yang & Liu 1996). Vibrio fluvialis, V. alginolyticus, V. cholerae non-O1 (Fujioka & Greco 1984), Aeromonas liquifaciens and V. anguillarum (Colorni 1985) have been isolated from the larvae of M. rosenbergii. A profound relationship between the abundance of members of the family Vibrionaceae and larval mortality (Singh 1990) and the predominance of Vibrio in eggs, larvae and post-larvae of M. rosenbergii (Hameed, Rahaman, Alagan & Yoganandhan 2003) was reported. The present paper reports the isolation, characterization, pathogenicity and antibiotic sensitivity of V. alginolyticus associated with M. rosenbergii larvae during an occurrence of severe mass mortality at the ninth larval stage.

Anti–white spot syndrome virus activity of Ceriops tagal aqueous extract in giant tiger shrimp Penaeus monodon

White spot syndrome virus (WSSV), the most contagious pathogen of cultured shrimp, causes mass mortality, leading to huge economic loss to the shrimp industry. The lack of effective therapeutic or prophylactic measures has aggravated the situation, necessitating the development of antiviral agents. With this objective, the antiviral activity in the aqueous extract of a mangrove plant Ceriops tagal in Penaeus monodon was evaluated. The Ceriops tagal aqueous extract (CTAE) was non-toxic to shrimps at 50 mg/ml when injected intramuscularly at a dosage of 10 lL/animal (0.5 mg/animal) and showed a protective effect against WSSV at 30 mg/ml when mixed with WSSV suspension at a 1:1 ratio. When the extract was administered along with the diet and the animals were challenged orally, there was a dose-dependent increase in survival, culminating in 100 % survival at a concentration of 500 mg/kg body weight/day. Neither hypertrophied nuclei nor the viral envelope protein VP28 could be demonstrated in surviving shrimps using histology and indirect immunofluorescence histochemistry (IIFH), respectively. To elucidate the mode of action, the temporal expression of WSSV genes and shrimp immune genes, including antimicrobial peptides, was attempted. None of the viral genes were found to be expressed in shrimps that were fed with the extract and challenged or in those that were administered CTAE-exposed WSSV. The overall results suggest that the aqueous extract from C. tagal can protect P. monodon from white spot syndrome virus infection.