National Status and Trends Bioeffects Program: field methods

Environmental quality indicators provide resource managers with information useful to assess coastal condition and scientifically defensible decisions. Since 1984, the National Oceanic and Atmospheric Administration (NOAA), through its National Status and Trends (NS&T) Program, has provided environmental monitoring data on chemical, physical, and biological indicators of coastal environments. The program has two major monitoring components to meet its goals. The Bioeffects Assessments Program evaluates the health of bays, estuaries, and the coastal zone around the nation using the Sediment Quality Triad technique that includes measuring sediment contaminant concentrations, sediment toxicity and benthic community structure. The Mussel Watch Program is responsible for temporal coastal monitoring of contaminant concentrations by quantifying chemicals in bivalve mollusks. The NS&T Program is committed to providing the highest quality data to meet its statutory and scientific responsibilities. Data, metadata and information products are managed within the guidance protocols and standards set forth by NOAA’s Integrated Ocean Observing System (IOOS) and the National Monitoring Network, as recommended by the 2004 Ocean Action Plan. Thus, to meet these data requirements, quality assurance protocols have been an integral part of the NS&T Program since its inception. Documentation of sampling and analytical methods is an essential part of quality assurance practices. A step-by–step summary of the Bioeffects Program’s field standard operation procedures (SOP) are presented in this manual.

Sediment quality triad assessment in Kachemak Bay: characterization of soft bottom benthic habitats and contaminant bioeffects assessment condensed. A NOAA/NOS/NCCOS/CCMA Special Report

A baseline environmental characterization of the inner Kachemak Bay, Alaska was conducted using standardized National Status and Trends Bioeffects Program methods. Three sites near the village of Port Graham were also sampled for comparison. Concentrations of over 120 organic and metallic contaminants were analyzed. Ambient toxicity was assessed using two bioassays. A detailed benthic community condition assessment was performed. Habitat parameters (e.g. depth, salinity, temperature, dissolved oxygen, sediment grain size, and organic carbon content) that influence species and contaminant distribution were also measured at each sampling site. The following is the synopsis of findings • Sediments were mostly mixed silt and sand with pockets of muddy zones. Organic compounds (PAHs, DDTs, PCBs, chlorinated pesticides) were detected throughout the bay but at relatively low concentrations. With some exceptions, metals concentrations were relatively low and probably reflect the input of glacial runoff. • Homer Harbor had elevated concentrations of metallic and organic contaminants. Concentrations of organic contaminants measured were five to ten times higher in the harbor sites than in the open bay sites. Tributyltin was elevated in Homer Harbor relative to the other areas. • There was no evidence of residual PAHs attributable to oil spills, outside of local input in the confines of the harbor. • The benthic community is very diverse. Specific community assemblages were distributed based on depth and water clarity. Species richness and diversity was lower in the eastern end of the bay in the vicinity of the Fox River input. Abundance was also generally lower in the eastern portion of the study area, and in the intertidal areas near Homer. The eastern portions of the bay are stressed by the sediment load from glacial meltwater. • Significant toxicity was virtually absent. • The benthic fauna at Port Graham contained a significant number of species not found in Kachemak Bay. • Selected metal concentrations were elevated at Port Graham relative to Kachemak Bay, probably due to local geology. Organic contaminants were elevated at a site south of the village.

Sediment Quality Triad assessment in Kachemak Bay: characterization of soft bottom benthic habitats and contaminant bioeffects assessment

A baseline environmental characterization of the inner Kachemak Bay, Alaska was conducted using the sediment quality triad approach based on sediment chemistry, sediment toxicity, and benthic invertebrate community structure. The study area was subdivided into 5 strata based on geophysical and hydrodynamic patterns in the bay (eastern and western intertidal mud flats, eastern and western subtidal, and Homer Harbor). Three to seven locations were synoptically sampled within each stratum using a stratified random statistical design approach. Three sites near the village of Port Graham and two sites in the footprint of a proposed Homer Harbor expansion were also collected for comparison. Concentrations of over 120 organic and metallic contaminants were analyzed. Ambient toxicity was assessed using two amphipod bioassays. A detailed benthic community condition assessment was performed. Habitat parameters (depth, salinity, temperature, dissolved oxygen, sediment grain size, and organic carbon content) that influence species and contaminant distribution were also measured at each sampling site. Sediments were mostly mixed silt and sand; characteristic of high energy habitats, with pockets of muddy zones. Organic compounds (PAHs, DDTs, PCBs, cyclodienes, cyclohexanes) were detected throughout the bay but at relatively low concentrations. Tributyltin was elevated in Homer Harbor relative to the other strata. With a few exceptions, metals concentrations were relatively low and probably reflect the input of glacial runoff. Relative to other sites, Homer Harbor sites were shown to have elevated concentrations of metallic and organic contaminants. The Homer Harbor stratum however, is a deep, low energy depositional environment with fine grained sediment. Concentrations of organic contaminants measured were five to ten times higher in the harbor sites than in the open bay sites. Concentration of PAHs is of a particular interest because of the legacy of oil spills in the region. There was no evidence of residual PAHs attributable to oil spills, outside of local input, beyond the confines of the harbor. Concentrations were one to ten times below NOAA sediment quality guidelines. Selected metal concentrations were found to be relatively elevated compared to other data collected in the region. However, levels are still very low in the scale of NOAA’s sediment quality guidelines, and therefore appear to pose little or no ecotoxicity threat to biota. Infaunal assessment showed a diverse assemblage with more than 240 taxa recorded and abundances greater than 3,000 animals m-22 in all but a few locations. Annelid worms, crustaceans, snails, and clams were the dominant taxa accounting for 63 %, 19%, 5%, and 7 % respectively of total individuals. Specific benthic community assemblages were identified that were distributed based on depth and water clarity. Species richness and diversity was lower in the eastern end of the bay in the vicinity of the Fox River input. Abundance was also generally lower in the eastern portion of the study area, and in the intertidal areas near Homer. The eastern portions of the bay are stressed by the sediment load from glacial meltwater. Significant toxicity was virtually absent. Conditions at the sites immediately outside the existing Homer Harbor facility did not differ significantly from other subtidal locations in the open Kachemak Bay. The benthic fauna at Port Graham contained a significant number of species not found in Kachemak Bay. Contaminant conditions were variable depending on specific location. Selected metal concentrations were elevated at Port Graham and some were lower relative to Kachemak Bay, probably due to local geology. Some organic contaminants were accumulating at a depositional site.

An assessment of polybrominated diphenyl ethers (PBDEs) in sediments and bivalves of the U.S. coastal zone

NOAA’s Mussel Watch Program was designed to monitor the status and trends of chemical contamination of U.S. coastal waters, including the Great Lakes. The Program began in 1986 and is one of the longest running, continuous coastal monitoring programs that is national in scope. NOAA established Mussel Watch in response to a legislative mandate under Section 202 of Title II of the Marine Protection, Research and Sanctuaries Act (MPRSA) (33 USC 1442). In addition to monitoring contaminants throughout the Nation’s coastal shores, Mussel Watch stores samples in a specimen bank so that trends can be determined retrospectively for new and emerging contaminants of concern. In recent years, flame retardant chemicals, known as polybrominated diphenyl ethers (PBDEs), have generated international concern over their widespread distribution in the environment, their potential to bioaccumulate in humans and wildlife, and concern for suspected adverse human health effects. The Mussel Watch Program, with additional funding provided by NOAA’s Oceans and Human Health Initiative, conducted a study of PBDEs in bivalve tissues and sediments. This report, which represents the first national assessment of PBDEs in the U.S. coastal zone, shows that they are widely distributed. PBDE concentrations in both sediment and bivalve tissue correlate with human population density along the U.S. coastline. The national and watershed perspectives given in this report are intended to support research, local monitoring, resource management, and policy decisions concerning these contaminants.

Breeding, age and growth of the freshwater shark Wallago attu (Bloch and Schneider) from the Dhir Beel of the Brahmaputra Basin, Assam, India

The Dhir Beel, one of the major live beels of the Brahmaputra Basin, Assam, has an area of 689 ha and situated in Dhubri district of Assam. The dominance of freshwater shark, Wallago attu (8.10%) in the beel is a striking feature. Restricted breeding of W. attu once a year from June to September was observed. The mean observed length was 37.5, 65.0, 84.5 and 99.0 cm in the 6th, 12th, 18th and 24th months of age respectively. The length growth coefficient (K), the asymptotic length (L infinity ), and the arbitrary origin of the growth curve (t omicron ), for W. attu were estimated to be 0.054484 per month, 136.16 cm and 0.0355 month respectively. The calculated life span (T infinity ) of the fish is 123.86 months (about 10 years). The weight growth parameters were estimated where the monthly growth coefficient (K), the asymptotic weight (W infinity) and the arbitrary origin of the growth curve (t omicron) were found to be 0.0743 per month, 7636.92 gram and 0.431908 month respectively. The length-weight relationship follows the cube law.

Hatching and rearing of gourami, (Osphronemus goramy, Lacepede) in the Polonnaruwa nursery

Gourami have been maintained as spawners in the ponds of the Fisheries Station at Polonnaruwa since 1957, for the production of fingerlings needed for the stocking of the inland waters. The procedure adopted for the rearing of these fingerlings (see Ling 1962, Indrasena, and de Silva, 1964, for details) is briefly descripted.

Genetic analysis of east and west coast populations of Penaeus monodon from India based on random amplified polymorphic DNA

The east and west coast populations of wild Penaeus monodon in India were genetically characterized by RAPD analysis using six highly polymorphic primers reported earlier. The average genetic similarities within populations, based on profiles generated by all the six primers, were 0.828 and 0.851 for the east and west coast populations, respectively, values with individual primers ranging from 0.744 to 0.889. The average genetic similarity between populations across all the primers was 0.774. The number of bands found to be polymorphic were 38 (51.35%) and 37 (50.68%) in the east and west coast populations, respectively. Primer 5 yielded the highest level of polymorphism (63.63%) in the east coast population whereas primer 3 yielded the lowest level of polymorphism (36.36%) in the west coast population. The study reveals the existence of genetic variation in P. monodon stocks providing scope for genetic improvement through selective breeding. It also provides baseline data for future work on population structure analysis of P. monodon.

Demonstration of some selected aquaculture technologies under farming system research in Jessore and Santahar, Bangladesh

BFRI evolved some selected aquaculture technologies viz. polyculture of carps in perennial ponds, monoculture of short cycled fish species (BFRI super strain) in seasonal ponds and prawn seed production through backyard hatchery system have been demonstrated under Farming System Research (FSR) component in Jessore and Santahar regions. Both polyculture of carps and monoculture of short cycled fish species technologies were tested in farmer's ponds in Kaium Kula village near Jessore town. In polyculture trials, seven species comprising of silver carp (Hypophthalmichthys molirrix), catla (Catla catla), rohu (Labeo rohita), grass carp (Ctenopharyngodon idellus), common carp (Cyprinus carpio), mrigal (Cirrhinus cirrhosus) and silver barb (Barbonymus gonionotus) were stocked @ 9,500 (ratio 6:2:4:2:1:5:5); 10,750 (ratio 6:2:4:2:1:5:5) and 12,000 (ratio 6:2:4:2:1:5:4) fish/ha respectively in ponds of T1, T2 and T3 having three replications of each. The mean highest fish production was 3,148 kg/ha in T3, followed by 2,899 kg/ha in T1 and 2,875 kg/ha in T2. Production of T3 was significantly different (P<0.05) than both T1 and T2, while there was no significant differences (P>0.05) between the production of T1 and T2. In case of trial of short cycled fish species, two treatments were tested: T1 (comprising of BFRI super strain of Nile tilapia, silver carp, common carp and silver barb; ratio 3:5:1:1) and T2 (having only BFRI super strain of Nile tilapia). Stocking density in both the treatments were same (20,000 fish/ha). In this trial average production was higher in T1 (2,743 kg/ha) than that of T2 (2,369 kg/ha) but the production figure in these two treatments was not significantly different (P>0.05). Demonstration of backyard prawn hatchery technology was tested at Santahar region of Bogra district, North-west part of Bangladesh. This hatchery consisted of three main components i) bio-filter, ii) rearing tank unit (chari) and iii) air blower/air pump unit. Plastic drum of 200-250 l capacity and cemented chari of 200-250 l capacity were used as bio-filter and larval rearing containers respectively. A 0.5 hp air blower with 6 aquarium air pump were used to operate the aeration system in the hatchery. Diluted sea water (10-12 ppt) made from brine solution (200-250 ppt) collected from salt-bed was used in the backyard hatchery system of hatching of eggs and rearing of larvae. Rearing of first stage zoea-larvae was reared in three rearing tanks following the stocking densities of 40, 50 and 60/l of water respectively. Production of post-larvae were 20±0.82, 22±1.12 and 28±1.63/liter of water in treatments I, II and III respectively in 38, 40 and 39 days rearing period.

Effects of black Bengal goat manure on growth and production of Genetically Improved Farmed Tilapia (GIFT) starin [sic], Oreochromis niloticus

In a goat-tilapia integrated farming system, the effect of Black Bengal goat manure on the growth and production of Oreochromis niloticus was studied at the Freshwater Substation, Shantahar, Bogra for 4.5 months. The stocking density used in three treatments were, 200 goats and 15,000 GIFT strain/ha (T1); 300 goats and 15,000 GIFT strain/ha (T2); and only 15,000 GIFT strain/ha (T3). The initial individual total length and weight of stocked tilapia were 7.6 cm and 11.34 g, respectively. Twelve ponds each having an area of 40 square meters were used for this trail [sic]. On one side of each pond goat shed was constructed and the space allocated for each goat was 0.75 m x 1.5 m. Newly weaned black Bengal goats of average weight 8.45 kg was used in the trial. Every morning the goats manure was swiped out in ponds through the fixed opening of bamboo made floor of goat shed. Water depth of the ponds was maintained at 0.75 m. The highest fish production was in treatment 1 (1,750 kg/ha) followed by treatment 2 (1,455 kg/ha) and treatment 3 (621 kg/ha). The difference in individual goat weight was not significant (P>0.05) among treatments 1 and 2.

Trophic State Index, morphoedaphic index and fish yield prediction in a sub-tropical lake, Manchar (Sindh), Pakistan

Limnological factors of a sub-tropical lake Manchar were studied on seasonal basis. The mean values of various parameters were: transparency, (secchi disc reading): 90.5 cm, Orthophosphate: 0.257 mg/l, TDS: 3310,5 mg/l, Conductivity: 5232 µs/l, Total Chlorophyll (Planktonic): 31.3 µg/l Planktonic biomass: 5466 µg/l. Trophic state index (TSI) was calculated by using Carlson's (1977) equations. Mean TSI for transparency was 61, while for orthophosphate and chlorophyll, it was 82 and 64 respectively. TSI values indicate advanced eutrophic state of Manchar Lake. Morphoedaphic index (MEI) was also calculated on seasonal basis. The mean values were, TDS: 1103, conductivity: 1744, alkalinity: 60, transparency: 29 and biomass (plankton dry weight): 1746. Fish yield prediction for Manchar Lake (Z =3m, mean area=100 km²) was calculated by using MEI values. The results were quite different among various parameters. Conductivity (89.1mt/y), biomass (67.6 mt/y) and TDS (44.6 mt/y) were found to be good predictors of fish yield. Chlorophyll, transparency and alkalinity values gave very low estimate.

Gross clinical signs and haematological changes associated with artifical infection of Edwardsiella tarda in Koi Carp

The occurrence of diseases is a significant setback for successful aquafarming. One of the common fish bacterial disease syndromes, Edwardsiellosis is caused by Edwardsiella tarda, a gram-negative, rod shaped bacterium associated with several diseases of marine and fresh water fish. In this study, an attempt was made to observe and analyze the onset of clinical symptoms and certain haematological parameters in Koi Carp, Cyprinus carpio L., following artificial infection with Edwardsiella tarda. The disease progress was observed and the clinical symptoms were monitored over a period of 15 days following infection. Fish were sampled at three day intervals to analyse the haematological parameters: total erythrocyte counts (RBC), total leucocyte counts (WBC), haemoglobin content and differential leucocyte count. Clinical symptoms observed included: erratic swimming behaviour, loss of appetite, haemorrhages, dropsy and exophthalmia. There was a significant decrease in the total RBC and haemoglobin levels by the 3rd and 6th day post infection, and an increase thereafter. WBC counts were higher in all infected groups in comparison to the control group. A significant increase in the number of neutrophils was found in the infected group up to the 9th day and a decrease thereafter. The lymphocyte number was significantly less up to the 12th day while the monocyte counts were significantly higher up to the 12th day post infection. The results showed that the bacterium, E. tarda, is pathogenic to Koi Carp. The hematological changes and clinical signs in infected fish reported in this paper will be helpful in the identification and the control of this infection.

Development of cell culture from caudal fin and heart of Tor tor (Hamilton-Buchanan)

Tor mahseer (Tor tor), possess high commercial and recreational value as they are potential game as well as food fish of India. Two cell culture systems were developed from fin and heart of T. tor (Hamilton-Buchanan). The explants excised aseptically from fingerling of T. tor were cultured in Leibovitz-15 (L-15) medium with 20% fetal bovine serum (FBS). Radiation of cells started after 72 hours and 48 hours of explant attachment from caudal fin and heart respectively. Confluent monolayer of cells with heterogeneous morphology around fin explants was observed after 7-10 days, where as a homogenous confluent layer of fibroblastic cells from heart explant was observed after 12-13 days. The establishment of cell culture systems from different organs and tissues of commercial important species would facilitates in vitro research.