Biofloc Technology: Emerging Microbial Biotechnology for the Improvement of Aquaculture Productivity

With the significant increases in the human population, global aquaculture has undergone a great increase during the last decade. The management of optimum conditions for fish production, which are entirely based on the physicochemical and biological qualities of water, plays a vital role in the prompt aquaculture growth. Therefore, focusing on research that highlights the understanding of water quality and breeding systems’ stability is very important. The biofloc technology (BFT) is a system that maximizes aquaculture productivity by using microbial biotechnology to increase the efficacy and utilization of fish feeds, where toxic materials such as nitrogen components are treated and converted to a useful product, like a protein for using as supplementary feeds to the fish and crustaceans. Thus, biofloc is an excellent technology used to develop the aquaculture system under limited or zero water exchange with high fish stocking density, strong aeration, and biota. This review is highlighted on biofloc composition and mechanism of system work, especially the optimization of water quality and treatment of ammonium wastes. In addition, the advantages and disadvantages of the BFT system have been explained. Finally, the importance of contemporary research on biofloc systems as a figure of microbial biotechnology has been emphasized with arguments for developing this system for better production of aquaculture with limited natural resources of water.Key words: biofloc, BFT, aquaculture, microbes, water quality, wastes     

Single nucleotide polymorphism discovery in albacore and Atlantic bluefin tuna provides insights into worldwide population structure

The optimal management of the commercially important, but mostly over‐exploited, pelagic tunas, albacore (Thunnus alalunga Bonn., 1788) and Atlantic bluefin tuna (BFT; Thunnus thynnus L., 1758), requires a better understanding of population structure than has been provided by previous molecular methods. Despite numerous studies of both species, their population structures remain controversial. This study reports the development of single nucleotide polymorphisms (SNPs) in albacore and BFT and the application of these SNPs to survey genetic variability across the geographic ranges of these tunas. A total of 616 SNPs were discovered in 35 albacore tuna by comparing sequences of 54 nuclear DNA fragments. A panel of 53 SNPs yielded F_ST values ranging from 0.0 to 0.050 between samples after genotyping 460 albacore collected throughout the distribution of this species. No significant heterogeneity was detected within oceans, but between‐ocean comparisons (Atlantic, Pacific and Indian oceans along with Mediterranean Sea) were significant. Additionally, a 17‐SNP panel was developed in Atlantic BFT by cross‐species amplification in 107 fish. This limited number of SNPs discriminated between samples from the two major spawning areas of Atlantic BFT (F_ST = 0.116). The SNP markers developed in this study can be used to genotype large numbers of fish without the need for standardizing alleles among laboratories.     

Characterization of three pro-inflammatory cytokines,TNFα1, TNFα2 and IL-1β, in cage-reared Atlantic bluefin tuna Thunnus thynnus

Atlantic bluefin tuna (BFT) (Thunnus thynnus) is of great economic significance for world aquaculture and therefore it is necessary to ensure optimal and sustainable conditions for the farming of this species. Intensive culture of fish may be limited by infectious diseases that can impact on growth performance and cause heavy losses. However, to date there are no reports of cloning and expression analysis of any major immune genes of Atlantic BFT although some immune genes are known in other BFT species. Therefore the aim of this study was to characterize the first cytokine molecules in Atlantic BFT, through: 1) Isolation of full-length cDNA and gene sequences of TNFα1, TNFα2 and IL-1β, 2) comparison of these molecules to known sequences in other vertebrates, especially teleost fish, by multiple sequence alignment, phylogenetic tree analysis and homology modeling; 3) Quantification of in vivo expression of these cytokines in selected tissues in reared BFT over the duration of the farming process. The results indicated that these three cytokines could have value for monitoring Atlantic BFT health status. Curiously, the liver seemed to be an important site of cytokine production during poor health conditions in this species, perhaps reflecting its role as an important organ involved in fish defenses.     

Filling the gap: Length–weight and length–length relationships of intertidal endemic fishes of the Brazilian Province Oceanic Islands

Length‐weight and length‐length relationships were determined for nine fish species of Rocas Atoll, Fernando de Noronha and Trindade Island. Samples were conducted in 2014 (Rocas) and 2015 (Noronha and Trindade) in tidepools using anaesthetic clove oil and hand nets to collect fish fauna. Four species (Stegastes rocasensis, Scartella itajobi, Starksia multilepis and Bathygobius brasiliensis) are endemic from the Noronha‐Rocas ridge, two are endemic from the Trindade‐Martin Vaz insular complex (Scartella poiti and Malacoctenus brunoi), and three species have a widespread distribution. All relationships are novel for science.     

DNA barcoding Neotropical fishes: recent advances from the Pampa Plain,Argentina

The fish fauna of the Pampa Plain, the southernmost distribution range of many Neotropical species, was barcoded in this study. COI sequences were analysed by means of distance (K2P/NJ) and character‐based (ML) models, as well as the Barcode Index Number (BIN). K2P/NJ analysis was able to discriminate among all previously identified species while also revealing the likely occurrence of two cryptic species that were further supported by BIN and ML analyses. On the other hand, both BIN and ML were not able to discriminate between two species of Rineloricaria. Despite the small genetic divergence between A. cf. pampa and A. eigenmanniorum, a tight array of haplotypes was observed for each species in both the distance and character‐based methods. Deep intraspecific divergences were detected in Cnesterodon decemmaculatus (5%) and Salminus brasiliensis (6%). For Salminus brasiliensis, these findings were further supported by character‐based (ML) evidence and meristic and morphological data. Our results also showed that Pampa Plain representatives of Salminus brasiliensis, Rhamdia quelen, Hoplias malabaricus, Synbranchus marmoratus, Australoheros facetus, Oligosarcus jenynsii and Corydoras paleatus differed by more than 3% from their conspecifics from other parts of South America. Overall, this study was able to highlight the likely occurrence of a cryptic species in Salminus brasiliensis and also illustrate the strong geographical structure in the COI sequence composition of seven fish species from South America.     

Identification of a sex‐specific molecular marker in Salminus brasiliensis (Characiformes) based on SCAR marker

Salminus brasiliensis, a characin regionally named ‘dourado’ (meaning goldfish), is the biggest characid fish, this species is cited in a list of fishes that are vulnerable. The goal of the present work was to try to identify, through AFLP technique to convert them into single locus markers (SCARs), in order to improve the sex identification in S. brasiliensis. Between possible sex‐specific AFLP markers in this study, one male‐specific were isolated (Les1) and converted into a SCAR marker. Les1 marker was confirmed sex‐specific in all samples tested. A sex‐specific DNA marker applicable to fish would be very useful for elucidating sex determination mechanism in fish.     

Changes in milk composition during lactation in three species of insectivorous bats

Changes in milk composition are described for three species of free-ranging insectivorous bats (Myotis lucifugus, M. velifer, and Tadarida brasiliensis) from early to mid (peak) lactation. Dry matter and energy concentrations in milk increased from early to mid-lactation. In M. lucifugus and T. brasiliensis, but not M. velifer, these increases were due largely to a rise in fat concentration, since protein and carbohydrate remained relatively constant. Energy content of milk (kJ·g^-1) for each species from early through mid-lactation was related to dry matter (DM) as follows: M. lucifugus (y=0.31 DM-0.32, r ²=0.68), M. velifer (y=0.48 DM-5.08, r ²=0.99), and T. brasiliensis (y=0.37 DM-1.51, r ²=0.61). Comparison of the effect of sampling method on milk composition of T. brasiliensis indicated that fat, dry matter, and energy concentrations increased significantly from pre-dawn to prenoon samples. Relatively high fat and low water levels in T. brasiliensis milk may reflect the limited access that lactating females have to free water, as well as need to minimize mass of stored milk during long foraging trips. Conversely, lower fat concentrations and higher water levels in milk in M. lucifugus and M. velifer may relate to the propensity for colonies of these two species to roost and forage near bodies of water. In addition, differences in milk fat concentrations observed among the three species may correlate to daily suckling schedules. Females of T. brasiliensis, for example, roost apart from and suckle their young on a regular daily schedule, whereas both species of Myotis roost with their pups and appear to suckle them on demand.Abbreviations bm body mass - DM dry matter - T _a air temperature     

Separating the effects of water quality and urbanization on temperate insectivorous bats at the landscape scale

Many local scale studies have shown that bats respond to water quality degradation or urbanization in a species‐specific manner. However, few have separated the effects of urbanization versus water quality degradation on bats, in single city or single watershed case studies. Across North Carolina, USA, we used the standardized North American Bat Monitoring Program mobile transect protocol to survey bat activity in 2015 and 2016 at 41 sites. We collected statewide water quality and urban land cover data to disentangle the effects of urbanization and water quality degradation on bats at the landscape scale. We found that statewide, water quality degradation and urbanization were not correlated. We found that bats responded to water quality degradation and urbanization independently at the landscape scale. Eptesicus fuscus and Lasiurus cinereus negatively responded to water quality degradation. Lasiurus borealis and Perimyotis subflavus positively responded to water quality degradation. Lasionycteris noctivagans did not respond to water quality degradation but was more active in more urbanized areas. Tadarida brasiliensis positively responded to urbanization and was less active in areas with degraded water quality. We show that bat–water quality relationships found at the local scale are evident at a landscape scale. We confirm that bats are useful bioindicators for both urbanization and water quality degradation. We suggest that water quality can be used to predict the presence of bat species of conservation concern, such as P. subflavus, in areas where it has not been studied locally.     

Parasites as biological tags for stock discrimination of the Brazilian flathead Percophis brasiliensis in the south-west Atlantic

Three stocks of the Brazilian flathead Percophis brasiliensis were identified on the coast of Argentina and Uruguay using parasites as biological tags. A total of 177 fish were examined and 23 parasite species were found. Fish were caught in four zones: north of the Argentine–Uruguayan Common Fishing Zone (34°30′–36°30′ S; 53°30′–56°00′ W), south of the Argentine–Uruguayan Common Fishing Zone (38°08′ S 57°32′ W), El Rincón zone (39–41° S; 60–62° W) and San Matías Gulf (41°40′–42°10′ S; 63°50′–65°00′ W). Discriminant analyses allowed the identification of three discrete stocks in the four zones (86·44% of classified samples were correctly identified), with P. brasiliensis from both north and south of the Argentine–Uruguayan Common Fishing Zone clumping together, as a single stock. Some species were important in discriminating among groups, Anisakis simplex was related to southern areas, while Grillotia sp., Corynosoma australe and Hysterothylacium sp. were important in determining the position of fish from the Argentine–Uruguayan Common Fishing Zone. These results were corroborated by comparing parasite prevalence and abundance among zones. The main differences were observed in those comparisons involving fish from the San Matías Gulf. Fish from the Argentine–Uruguayan Common Fishing Zone were characterized by higher infections of Grillotia sp., C. australe, Corynosoma cetaceum and Hysterothylacium sp., while the samples from El Rincón and San Matías Gulf showed higher infections of A. simplex. Samples from San Matías Gulf were characterized by lower levels of parasitism for all other species. Differences in environmental factors and their influence on the distribution of zooplankton and other hosts in the food web may be differentially shaping the parasite community structure in each zone, resulting in identifiable stocks of the P. brasiliensis. The present study confirmed the existence of regional biological tags that delineated fish assemblages.     

Restoration of Zooplankton Communities in Industrially Damaged Lakes: Influences of Residual Metal Contamination and the Recovery of Fish Communities

The Sudbury, Ontario, Canada area offers a unique opportunity to develop our understanding of biotic and abiotic lake recovery processes in industrially damaged natural systems. In recent decades, lakes in the Sudbury area have shown improvements in water quality due to decreases in sulfur (S) and metal emissions from area smelters, and reduced acid deposition from more distant sources. However, biological recovery is lagging and mechanisms controlling the lag are not yet clear. Our study examines the roles of two factors, residual metal contamination and altered fish predation, on zooplankton community recovery. Data collected over three decades on six study lakes were analyzed using redundancy analysis (RDA) and partial RDA's to assess historical and present relationships of water chemistry and fish abundance with zooplankton community recovery. Continuing metal toxicity appears to be the primary cause of the absence of some zooplankton species, particularly Daphnia spp. from metal‐contaminated lakes. Conversely, once water quality is suitable and abundant planktivores reestablish, fish planktivory becomes a factor affecting Daphnia spp. establishment. The introduction of piscivores into these lakes may be necessary to facilitate the return of many Daphnia species. Further reductions in metal toxicity will also assist with the complete recovery of zooplankton communities. Studying natural systems over several decades allows us to better understand the intricate steps involved with recovery of industrially damaged lakes, and this knowledge will greatly benefit future restoration efforts in other industrially damaged systems .     

Influence of environmental variables on the growth and reproductive cycle of Crassostrea (Mollusca,Bivalvia) in Guaratuba Bay,Brazil

The production of oysters in Guaratuba Bay, in the state of Paraná, Brazil, is still a mixed activity of mariculture and extractivism. The sustainable development of this production requires the monitoring of environmental, genetic, reproductive, and zootechnical variables. This study evaluated the importance of these variables on Crassostrea oyster production. Data were obtained between September 2009 and February 2011 from six evaluations: water quality, oyster larvae in plankton, capture of oyster spat by artificial collectors, molecular identification of collected spat and larvae, continuous evaluation of the reproductive maturity stage of adult oysters, and zootechnical performance achieved by experimental production. Temperature, dissolved oxygen, and water transparency were the environmental factors that had the most influence on the reproductive cycle (e.g. gonadal development and spat capture) and on the growth of the oyster. The highest rates for spat uptake were recorded in the summer, coinciding with the peak of sexual maturity of adults. The growth of the oysters, in terms of the weight of the meat, was related to the gonadal stage.     

Seasonal and interannual effects of hypoxia on fish habitat quality in central Lake Erie

1. Hypoxia occurs seasonally in many stratified coastal marine and freshwater ecosystems when bottom dissolved oxygen (DO) concentrations are depleted below 2–3 mg O₂ L^?1. 2. We evaluated the effects of hypoxia on fish habitat quality in the central basin of Lake Erie from 1987 to 2005, using bioenergetic growth rate potential (GRP) as a proxy for habitat quality. We compared the effect of hypoxia on habitat quality of (i) rainbow smelt, Osmerus mordax mordax Mitchill (young‐of‐year, YOY, and adult), a cold‐water planktivore, (ii) emerald shiner, Notropis atherinoides Rafinesque (adult), a warm‐water planktivore, (iii) yellow perch, Perca flavescens Mitchill (YOY and adult), a cool‐water benthopelagic omnivore and (iv) round goby Neogobius melanostomus Pallas (adult) a eurythermal benthivore. Annual thermal and DO profiles were generated from 1D thermal and DO hydrodynamics models developed for Lake Erie’s central basin. 3. Hypoxia occurred annually, typically from mid‐July to mid‐October, which spatially and temporally overlaps with otherwise high benthic habitat quality. Hypoxia reduced the habitat quality across fish species and life stages, but the magnitude of the reduction varied both among and within species because of the differences in tolerance to low DO levels and warm‐water temperatures. 4. Across years, trends in habitat quality mirrored trends in phosphorus concentration and water column oxygen demand in central Lake Erie. The per cent reduction in habitat quality owing to hypoxia was greatest for adult rainbow smelt and round goby (mean: ?35%), followed by adult emerald shiner (mean: ?12%), YOY rainbow smelt (mean: ?10%) and YOY and adult yellow perch (mean: ?8.5%). 5. Our results highlight the importance of differential spatiotemporally interactive effects of DO and temperature on relative fish habitat quality and quantity. These effects have the potential to influence the performance of individual fish species as well as population dynamics, trophic interactions and fish community structure.     

Arms as areas for larval development of migratory fish species in a Neotropical reservoir and the influence of rainfall over abundances

The aim of the present study was to verify the use of the arms of the Itaipu Reservoir as areas of initial development for migratory fish species and to assess the relationship between rainfall and the spawning of migratory fish. Accordingly, fish larvae were collected from five arms of the reservoir from 2009 to 2016 using 0.5 mm plankton nets. Density was standardized as the number of larvae per 10 m³ filtered water, and the captured larval and juvenile specimens were identified at the lowest-possible taxonomic level. The larvae were also classified according to the degree of development and notochord flexion stage: larval vitelline, pre-flexion, flexion and post-flexion. To evaluate the distribution of larval abundance and the developmental stage along the longitudinal gradients of the arms, the data were evaluated using a set of nested linear models, following the AIC and Bayesian information criteria. In addition, an analysis of covariance was performed to investigate the influence of rainfall on the larval abundance of migratory species. During sampling, several species of economic and conservation interest such as Salminus brasiliensis and Pseudoplatystoma corruscans were collected. The larvae of the migratory fish taxa were captured from all sampled arms, which indicate them as areas of initial development. Nevertheless, it was observed that larval density increases from fluvial towards lacustrine zones inside the arms. Also, the present study verified that species, even in lentic environments, respond positively to rainfall stimuli in a manner similar to that exhibited by conspecifics in lotic environments. Such results reinforce the necessity of the protection of arms aiming at the conservation of this main group of species impaired by the construction of dams.     

Mollugo viscosa sp. nov. (Molluginaceae), a segregate of M. brasiliensis from Minas Gerais in eastern Brazil

The new species Mollugo viscosa Thulin & Harley is described from Minas Gerais in eastern Brazil. It was previously part of the recently published M. brasiliensis Thulin & Harley, a species that was found to be non‐monophyletic in molecular phylogenetic analyses. Mollugo brasiliensis is here divided into a northern species (M. brasiliensis s.str.) in Bahia and a southern species (M. viscosa sp. nov.) in Minas Gerais. The new species differs from M. brasiliensis in its denser and more viscous indumentum, and in its distinctly smaller seeds with smaller tubercles and a smaller hilar peg, and from its sister species M. ulei by being a perennial herb with bisexual flowers.     

Serological Survey of Paracoccidioidomycosis in Sheep

The objective of this study was to evaluate the prevalence of antibodies against Paracoccidioides brasiliensis in sheep from Guarapuava, Paraná State, Brazil. The seroepidemiological study was carried out in 262 sheep. The samples were analyzed by ELISA and immunodiffusion test using P. brasiliensis gp43 and exoantigen as antigens, respectively. Initially, two sheep were immunized with P. brasiliensis to evaluate whether contact with the fungal cells could induce a humoral immune response against gp43 and exoantigen from P. brasiliensis. Both animals produced antibodies against gp43 and exoantigen, the main antigens used for diagnosis and seroepidemiology of paracoccidioidomycosis. A reactivity of 37% was observed to the P. brasiliensis gp43 antigen by ELISA although no reactivity had been observed by the immunodiffusion test. Sheep under extensive grazing system showed higher frequency of positivity to P. brasiliensis (P ≤ 0.05) than those under intensive and semi-intensive systems. These data suggest that sheep may be a useful epidemiological marker of P. brasiliensis presence in the environment and reinforce that contact with soil is an important risk factor for infection.     

Use of horizontal subsurface flow constructed wetlands to treat reverse osmosis concentrate of rolling wastewater

According to the characteristics of the reverse osmosis concentrate (ROC) generated from iron and steel company, we used three sets of parallel horizontal subsurface flow (HSF) constructed wetlands (CWs) with different plants and substrate layouts to treat the high-salinity wastewater. The plant growth and removal efficiencies under saline condition were evaluated. The evaluation was based entirely on routinely collected water quality data and the physical and chemical characteristics of the plants (Phragmites australis, Typha latifolia, Iris wilsonii, and Scirpus planiculmis). The principal parameters of concern in the effluent were chemical oxygen demand (COD), total nitrogen (TN), and total phosphorus (TP). The results showed that the CWs were able to remove COD, TN, and TP from ROC. S. planiculmis was not suitable for the treatment of high-saline wastewater. The sequence of metals accumulated in CW plants was K>Ca>Na>Mg>Zn>Cu. More than 70% of metals were accumulated in the aboveground of P. australis. The CW filled with gravel and manganese ore and planted with P. australis and T. latifolia had the best performance of pollutant removal, with average removal of 49.96%, 39.45%, and 72.01% for COD, TN, and TP, respectively. The effluent water quality met the regulation in China. These results suggested that HSF CW planted with P. australis and T. latifolia can be applied for ROC pollutants removal.     

Environmental influences on the replenishment of lizardfish (family Synodontidae) in Caribbean Panama

Lizardfish (family Synodontidae) are little studied despite their potentially important predatory role in epibenthic coral reef communities. The present study documented the temporal and spatial larval supply patterns of five Caribbean lizardfish species together with environmental variables (solar radiation, rainfall, water temperature, onshore-offshore winds, alongshore winds and wind-induced turbulence) to examine: (1) whether species within the same family respond differently to their developmental environment and (2) if larval supply in year-round warm waters is influenced by climatic changes. To address these questions, late-stage larvae of Synodus foetens, Synodus intermedius, Synodus poeyi, Saurida suspicio and Saurida brasiliensis were collected in replicate light traps in three different reef habitats (back-reef, lagoon and exposed) in the San Blas Archipelago, Panama, over 18 consecutive lunar months. Although replenishment of lizardfish occurred year-round, the temporal and spatial supply patterns were species-specific: S. foetens, S. intermedius and S. poeyi were most abundant during the dry season while S. suspicio and S. brasiliensis were most prevalent during the wet season. When seasons were analysed separately, water temperature explained 39 and 26% of the variance in light trap catches of S. foetens and S. intermedius, respectively, in the dry season while wind-induced turbulence accounted for 25% of the variability in S. suspicio and S. brasiliensis catches during the wet season. These findings stress the importance of analysing larval supply in conjunction with environmental data at a high taxonomic resolution to better understand the mechanisms that drive replenishment in reef systems at low latitudes.     

Constructed Wetlands for Tannery Wastewater Treatment in Portugal: Ten Years of Experience

Wastewaters from tannery industry are complex in composition and providing adequate treatment can be difficult. Constructed wetlands (CW) are regarded as an alternative treatment to the conventional biological systems, as a developing cost-effective and environmentally friendly phytoremediation technology. The present review compiles and integrates information on CWs technology for the needs of the tannery sector. The following issues arise as crucial for the implementation of such systems, namely i) an accurate wastewater characterization and an effective pretreatment before reaching the CW, ii) choosing the plants species better adapted to the imposed conditions, iii) substrate selection and iv) range of organic loadings applied. The examples practiced in Portugal give indication that horizontal subsurface flow systems, with expanded clay media, are a suitable option to be considered when dealing with high organic loading tannery wastewater (up to c.a. 3800 kgCODha^?1d^?1), being resilient to a wide range of hydraulic variations. Plants such as Phragmites and Typha have shown to be adequate for tannery wastewater depuration, with Arundo donax proving resilient to high salinity wastewaters. The flexibility of implementation allows the CW to be adapted to different sites with different configurations, being suitable as main secondary or tertiary treatment stage.     

Length–weight relationships for 25 fish species from three coastal lagoons in Southeastern Brazil

This work analyzes the relationship between length and weight for 25 fish species belonging to 15 families in three mixohaline/hypersaline coastal lagoons in Southeastern Brazil. The study presents the first estimation of L–WRs for six species (Anchoa tricolor, Brevoortia aurea, Jenynsia multidentata, Ctenogobius boleosoma, Microgobius meeki and Bathygobius soporator) and maximum lengths for four species (Atherinella brasiliensis, Jenynsia multidentata, Poecilia vivipara and Microgobius meeki) that are greater than previously recorded.     

Response of stream fish to improved water quality: a comparison between the 1960s and 1990s

1. The effect of improved water quality on fish assemblages in streams in southern Sweden was assessed by comparing species composition at 161 sites and water quality at twenty-nine sites in the 1960s and the 1990s. 2. Water quality had improved and there was an increase in the number of sites or catchments with brown trout (Salmo trutta), stone loach (Barbatula barbatula) and eel (Anguilla anguilla). The response was greatest for brown trout and was best explained by increased oxygen concentrations. 3. The number of sites with nine-spined stickleback (Pungitius pungitius), ide (Leuciscus idus) and brook lamprey (Lampetra planeri) decreased between the 1960s and 1990s. The decrease was greatest for nine-spined stickleback and was related to the increase in sites with trout, suggesting that nine-spined stickleback may be sensitive to predation or competition. 4. Improved water quality has led to recolonization by brown trout, probably enabling biotic interactions to play a larger role in structuring fish assemblages. Improving water quality was an effective method for rehabilitating fish populations in streams where natural colonization was possible.