Progress 09/15/05 to 09/14/08
Outputs
OUTPUTS: Biofiltration: The biofiltration studies of normal operating conditions were completed and included in an MS Thesis in the department of Biological and Agricultural Engineering. Results of this study have also been submitted for publication in two journal manuscripts. Findings from this study have been presented at a three conferences and 4 extension workshops in the US, Spain and Australia. Knowledge gained in this project has been used in the design of large scale aquaculture facilities in NC and Florida. Sex Determination in Flounder: The southern flounder studies were completed as proposed. Results of this work are included in a MS Thesis in the department of Zoology. Results of this work were presented at in a series of 4 workshops on southern flounder culture and at international research conferences in Brazil and the USA. Compensatory Growth: We conducted a compensatory growth tank study, where animals are fed following a period of feed restriction, to produce hybrid striped bass that exhibit poor, normal and supra-accelerated growth rates and to determine if feeding manipulation might improve production characteristics of this important cultivar. We measured the blood and tissue protein levels and gene expression of various growth regulatory hormones and their receptors and to establish how they might regulate differential growth rates. We developed blood insulin-like growth factor-I measures as an effective indicator of growth status of hybrid striped bass. Results of this work have been disseminated to producers and scientists through various mechanisms that include publications, five presentations at the annual and triennial meetings of the World Aquaculture Society, American Fisheries Society, and North Carolina Aquaculture Development conferences, and through direct on farm research activities with producers. Striped Bass Egg Quality: We conducted an investigation of the molecular determinants of egg quality in temperate basses (genus Morone) based on the hypothesis that disruption of a system of multiple egg yolk precursors, vitellogenins (Vtgs) is a leading cause of poor egg quality in these species. Three complete cDNAs encoding different forms of white perch (Morone americana) Vtgs were isolated and characterized with respect to immuno-biochemical and functional features of the three Vtg proteins and their product yolk proteins (YPs) in this species and in the congeneric striped bass (M. saxatilis). We have collected biopsy samples of oocytes and ovulated eggs from female striped bass undergoing final maturation and yielding good and poor quality eggs based on fertility and embryo/fry survival estimates. Results of this work have been disseminated to producers and scientists through several publications in per reviewed journals, 5 presentations at the 8th International Symposium on the Reproductive Physiology of Fish, a presentation at the 3rd International Symposium on Viviparous Fishes, a presentation at the triennial meeting of the WAS, a seminar at the NC Aquaculture Conference, and through direct contact with hybrid striped bass growers. PARTICIPANTS: Biofiltration: Individuals: Thomas Losordo, (PI, NC State University) Scientific Director; Dennis DeLong, (Extension Specialist, NC State University, onsite project manager; Richard Jones, (Research Technician, NC State University) onsite technical assistant; Todd Guerdat (Graduate Research Assistant, NC State Univeristy) conducted research activities on this project. Collaborators: Harry Daniels, NC State University; Kevan Main, Mote Aquaculture, Saratota, Florida. Partner Organizations: Amy Riedel (Aquatic EcoSystems, Inc) coordinated workshop in Florida. Flounder: Individuals: Harry Daniels, (PI, NC State University)Scientific Director, Ryan Murashige (Research Technician, NC State University) onsite project management, Robin Sanderson, (Research Technician, NC State University) onsite project management. Collaborators: Wade Watanabe, University of NC Wilmington Russell Borski, NC State University John Godwin, NC State University. Partner Organizations: Keith Hairr, Owner, Carolina Flounder, LLC Golden LEAF Foundation. Compensatory Growth: Individuals: Russell Borski (PI, NC State University) managed the project. Matthew Picha (Ph.D. graduate student), Eugene Won (M.S. graduate student) and Marc Turano (Ph.D. graduate student), Christian Tipsmark (Postdoctoral Fellow), Jennifer Schmidt (undergraduate student), conducted the research activities, analyses and other work for this and related research. Partner Organizations: William Watson (Owner Westside Fisheries - Hybrid Striped Bass Grower); NC Sea Grant; NC State University. Collaborators and contacts; Harry Daniels (Professor, North Carolina State University) Training or professional development: The research outlined in the proposed studies provided the research training and experiences for 1 Postdoctoral Fellow, 3 graduate students, and one undergraduate student and work with a hybrid striped bass producer (see individuals and partner organizations). Egg Quality: Individuals: Craig Sullivan (Principal Investigator, NC State University) managed the project. Naoshi Hiramatsu (Postdoctoral Fellow), Kaori Hiramatsu (Research Specialist), Benjamin Reading (Ph.D. graduate student), Sandra Page (Ph.D. graduate Student), Amanda Boury (M.Sc. graduate student), Valerie Williams (undergraduate student), and Cherie Gothreau (undergraduate student) conducted the research activities, analyses and other work for this and related research. Partner Organizations: Hybrid striped bass growers Lee H. Brothers (Owner, Carolina Fisheries), Scott Austin (Owner, Austin Brothers Fisheries), and Steven Locke (President, CastleHayne Fisheries); North Carolina Sea Grant; Pamlico Aquaculture Field Laboratory, NC State University. Collaborators and contacts: Akihiko Hara (Dean, Faculty of Fisheries, Hokkaido University, Japan)Takahiro Matsubara (Assistant Director, National Fisheries Research Institute, Kushiro, Japan) E. Gordon Grau (Professor, Hawai Institute of Marine Biology, University of Hawaii). Training or professional development: The research outlined in the proposed studies provided the research training and experiences for 1 Postdoctoral Fellow, 3 graduate students, and two undergraduate students. TARGET AUDIENCES: Biofiltration: National and international aquaculture producers using recirculating systems. Workshops on recirculating systems at the North Carolina Aquaculture Conference and with a cooperator in Florida. Flounder: U.S. aquaculture producers. Workshops on southern flounder culture. Presentations at producer meetings and annual North Carolina Aquaculture Development Conference. Compensatory Growth: This research targeted individuals in aquaculture sciences, the aquaculture industry, and to producers of hybrid striped bass. Results of this work have been disseminated to producers and scientists through various mechanisms that include publications, five presentations at the annual and triennial meetings of the World Aquaculture Society, American Fisheries Society, and North Carolina Aquaculture Development conferences, four seminar presentations at various national and international universities and aquaculture institutes, and through direct on farm research activities with hybrid striped bass producers in North Carolina (Westside Fisheries). Egg Quality: This research targeted individuals, including students, in aquaculture sciences, the general aquaculture industry, and producers of hybrid striped bass nationwide and especially in eastern North Carolina. Results of this work have been disseminated to producers and scientists and students through various mechanisms that include scholarly publications, multiple presentations at international, national and State symposia and conferences dealing with aquaculture issues, direct instruction of graduate and undergraduate students in courses taught by C.V. Sullivan at N.C. State University (ZO 586/587 Aquaculture with Laboratory; ZO 515 Fish Physiology), hands on training of postdoctoral, graduate and undergraduate students during execution of the research, and direct contact with Hybrid striped bass growers during research conducted, in part, on their farms. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
Biofiltration: Prior to this study, the vast majority of biofilter nitrification rate information was created in laboratory scale filters, often with artificial wastes. This study provides a rare glimpse at actual biofilter nitrification rates in commercial scale under real aquaculture conditions. Total ammoniacal nitrogen (TAN) removal rates were determined for the three types of biofilters for a range of concentrations ranging from 0.13 to 1.20 g TAN m-3. Average observed TAN removal rates (in g TAN/ m3 of unexpanded media per day (standard deviation)) for the three filters were 267.2 (123.3), 586.3 (284.2), and 667.4(344.8)for the moving bed bioreactor, floating bead filter, and fluidized sand filter, respectively. These results are considerably lower than results previously published at the laboratory scale using artificial waste nutrients. This study highlights the need for future biofilter evaluations at the commercial scale using real aquaculture waste nutrients. Flounder: Three studies were conducted. The first study examined the effect of tank background color on sex ratios and cortisol concentrations in southern flounder. Southern flounder juveniles were reared in grey, black and blue tanks for 121 days. Blue tanks resulted in a significantly higher percentage of males than the other treatments. The second study looked at the effect of light intensity on sex ratios and cortisol concentrations. Juveniles were reared in three different light intensities (5, 100, 1500 lux) for 144 days. The 5 lux treatment resulted in a significantly lower percentage of males than the other two treatments. A third study examined the feasibility of feeding cortisol treated feed to juveniles. The juveniles were fed cortisol treated feed with 0, 100, and 500 mg/kg for 48 hours. Cortisol levels in treatment fish remained significantly elevated over control fish from 30 minutes post treatment. Compensatory Growth: We found that CG can be induced in hybrid striped bass (HSB) fed daily after 3-week periods of no feeding. The CG response is characterized by increases in specific growth rate (SGR), hyperphagia and improved feed conversion. Insulin-like growth factor-I (IGF-I) is a critical growth regulator in fish and the IGF binding proteins (IGFBPs) prolong IGF-I's half-life and regulates its availability to target tissues. During the feed restriction period, circulating insulin-like growth factor-I (IGF-I) and the IGFBP-3 declines in parallel with SGR. Upon refeeding, IGF-I and IGFBP-3 rapidly increase along with SGR. Overall, significant correlations between SGR and plasma IGF-I were seen during periods of slow, normal and accelerated growth in these experiments. Based on this work we conclude that IGFBP-3 may serve to prolong the growth promoting actions of IGF-I during states of rapid growth. Striped Bass Egg Quality: Working with the staff of the NCSU genome research laboratory we have pioneered a new mass spectroscopy method for detecting aberrant yolk protein processing in striped bass and related species that will contribute to our assessments of the physiological underpinnings of poor egg quality in these and other farmed species.
Publications
- Turano, M.J., Borski, R.J., and Daniels H.V. (2008) Effects of cyclic feeding on compensatory growth of hybrid striped bass (Morone chrysops x M. saxitilis) foodfish and water quality in production ponds. Aquaculture Research. 39:1514-1523.
- Amano, H., Fujita, T., Hiramatsu, N., Kagawa, H., Sawaguchi, S., Matsubara, T., Sullivan, C.V., and A. Hara. 2007. Molecular alteration of three forms of vitellogenins and their derivative yolk proteins during oocyte growth and maturation in grey mullet (Mugil cephalus). Cybium, the International Journal of Ichthyology (in press). Cybium 2008, 32(2) suppl.: 156-158.
- Davis, L., Hiramatsu, N., Sullivan, C.V., Hirano, T., and E.G. Grau. 2007. Estrogen regulation of multiple vitellogenin and estrogen receptor genes and of the growth hormone-insulin like growth factor axis in tilapia. Cybium, the International Journal of Ichthyology Cybium 2008, 32(2) suppl.: 242-243.
- Hiramatsu, N., Mihoka, I., Ideuchi, H., Fujita, T., Amano, H., Matsubara, T., Sullivan, C.V., and A. Hara. 2007. Differential production and uptake of dual vitellogenins in Japanese medaka (Oryzias latipes). Cybium, the International Journal of Ichthyology Cybium 2008, 32(2) suppl.: 260.
- Reading, B.J., Hiramatsu, N., Matsubara, T., Hara, A., and C.V. Sullivan. 2007. Deduced primary structures of three vitellogenins and specific binding to multiple ovarian receptors in white perch (Morone americana). Cybium, the International Journal of Ichthyology Cybium 2008, 32(2) suppl.: 159-161.
- Sawaguchi, S., Ohkubo, N., Amano, H., Hiramatsu, N., Hara, A., Sullivan, C.V., and T. Matsubara. Controlled accumulation of multiple vitellogenins into oocytes during vitellogenesis in the barfin flounder, Varasper moseri. Cybium, the International Journal of Ichthyology Cybium 2008, 32(2) suppl.: 262.
- Turner, P.M. 2008. Effects of Light Intensity and Tank Background Color on Sex Determination in Southern Flounder (Paralichthys lethostigma). M.Sc. Thesis. North Carolina State University, Raleigh, North Carolina.
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Progress 09/15/06 to 09/14/07
Outputs
OUTPUTS: Biofiltration: The proposed biofiltration studies are nearing completion. This project has been part of a MS graduate degree program. This project has provided an opportinuty for learning at the graduate level. Results of this study have been presented in writing and by an oral presentation by the student in a research seminar in his academic department. Ongoing findings from this study have been presented at workshops in NC and Florida. Knowledge gained in this project has been used in the design of a large scale aquaculture facility in NC and an aquaculture laboratory in Florida. Flounder: The proposed southern flounder studies have been completed as described. Results of this work are being presented in a series of workshops on southern flounder culture. We have conducted four of these workshops to current and prospective producers who are involved in the production of southern flounder. Compensatory Growth: We conducted a compensatory growth tank study, where animals are
fed following a period of feed restriction, to produce hybrid striped bass that exhibit poor, normal and supra-accelerated growth rates and to determine if feeding manipulation might improve production characteristics of this important cultivar. We measured the blood and tissue protein levels and gene expression of various growth regulatory hormones and their receptors and to establish how they might regulate differential growth rates. We developed blood insulin-like growth factor-I measures as an effective indicator of growth status of hybrid striped bass. Results of this work have been disseminated to producers and scientists through various mechanisms that include publications, five presentations at the annual and triennial meetings of the World Aquaculture Society (WAS), American Fisheries Society (AFS), and NC Aquaculture conferences, four seminar presentations at various national and international universities and aquaculture institutes, and through direct on farm research
activities with producers in NC (Westside Fisheries). Egg Quality: We conducted an investigation of the molecular determinants of egg quality in temperate basses (genus Morone) based on the hypothesis that disruption of a system of multiple egg yolk precursors, vitellogenins (Vtgs) is a leading cause of poor egg quality in these species. Basic investigations of the multiple Vtg system were conducted to discover the number, characteristics, and function of each form of egg yolk precursor in these species. Striped bass broodstock were sampled throughout their reproductive cycle to measure blood profiles of the different forms of Vtg and ovarian levels of the yolk proteins. Results of this work have been disseminated to producers and scientists through several publications in per reviewed journals, 5 presentations at the 8th International Symposium on the Reproductive Physiology of Fish, a presentation at the 3rd International Symposium on Viviparous Fishes, a presentation at the
triennial meeting of the WAS, a seminar at the NC Aquaculture Conference, and through direct contact with hybrid striped bass growers. (Carolina Fisheries, CastleHayne Fisheries, Pungo Fisheries, Austin Brothers Fisheries).
PARTICIPANTS: Biofiltration: Individuals: Thomas Losordo, (PI, NC State University) Scientific Director; Dennis DeLong, (Extension Specialist, NC State University, onsite project manager; Richard Jones, (Research Technician, NC State University) onsite technical assistant; Todd Guerdat (Graduate Research Assistant, NC State Univeristy) conducted research activities on this project. Collaborators: Harry Daniels, NC State University; Kevan Main, Mote Aquaculture, Saratota, Florida. Partner Organizations: Amy Riedel (Aquatic EcoSystems, Inc) coordinated workshop in Florida. Flounder: Individuals: Harry Daniels, (PI, NC State University)Scientific Director, Ryan Murashige (Research Technician, NC State University) onsite project management, Robin Sanderson, (Research Technician, NC State University) onsite project management. Collaborators: Wade Watanabe, University of NC Wilmington Russell Borski, NC State University John Godwin, NC State University. Partner Organizations: Keith Hairr,
Owner, Carolina Flounder, LLC Golden LEAF Foundation. Compensatory Growth: Individuals: Russell Borski (PI, NC State University) managed the project. Matthew Picha (Ph.D. graduate student), Eugene Won (M.S. graduate student) and Marc Turano (Ph.D. graduate student), Christian Tipsmark (Postdoctoral Fellow), Jennifer Schmidt (undergraduate student), conducted the research activities, analyses and other work for this and related research. Partner Organizations: William Watson (Owner Westside Fisheries - Hybrid Striped Bass Grower); NC Sea Grant; NC State University. Collaborators and contacts; Harry Daniels (Professor, North Carolina State University) Training or professional development: The research outlined in the proposed studies provided the research training and experiences for 1 Postdoctoral Fellow, 3 graduate students, and one undergraduate student and work with a hybrid striped bass producer (see individuals and partner organizations). Egg Quality: Individuals: Craig Sullivan
(Principal Investigator, NC State University) managed the project. Naoshi Hiramatsu (Postdoctoral Fellow), Kaori Hiramatsu (Research Specialist), Benjamin Reading (Ph.D. graduate student), Sandra Page (Ph.D. graduate Student), Amanda Boury (M.Sc. graduate student), Valerie Williams (undergraduate student), and Cherie Gothreau (undergraduate student) conducted the research activities, analyses and other work for this and related research. Partner Organizations: Hybrid striped bass growers Lee H. Brothers (Owner, Carolina Fisheries), Scott Austin (Owner, Austin Brothers Fisheries), and Steven Locke (President, CastleHayne Fisheries); North Carolina Sea Grant; Pamlico Aquaculture Field Laboratory, NC State University. Collaborators and contacts: Akihiko Hara (Dean, Faculty of Fisheries, Hokkaido University, Japan)Takahiro Matsubara (Assistant Director, National Fisheries Research Institute, Kushiro, Japan) E. Gordon Grau (Professor, Hawai Institute of Marine Biology, University of
Hawaii). Training or professional development: The research outlined in the proposed studies provided the research training and experiences for 1 Postdoctoral Fellow, 3 graduate students, and two undergraduate students.
TARGET AUDIENCES: Biofiltration: National and international aquaculture producers using recirculating systems. Workshops on recirculating systems at the North Carolina Aquaculture Conference and with a cooperator in Florida. Flounder: U.S. aquaculture producers. Workshops on southern flounder culture. Presentations at producer meetings and annual North Carolina Aquaculture Development Conference. Compensatory Growth: This research targeted individuals in aquaculture sciences, the aquaculture industry, and to producers of hybrid striped bass. Results of this work have been disseminated to producers and scientists through various mechanisms that include publications, five presentations at the annual and triennial meetings of the World Aquaculture Society, American Fisheries Society, and North Carolina Aquaculture Development conferences, four seminar presentations at various national and international universities and aquaculture institutes, and through direct on farm research
activities with hybrid striped bass producers in North Carolina (Westside Fisheries). Egg Quality: This research targeted individuals, including students, in aquaculture sciences, the general aquaculture industry, and producers of hybrid striped bass nationwide and especially in eastern North Carolina. Results of this work have been disseminated to producers and scientists and students through various mechanisms that include scholarly publications, multiple presentations at international, national and State symposia and conferences dealing with aquaculture issues, direct instruction of graduate and undergraduate students in courses taught by C.V. Sullivan at N.C. State University (ZO 586/587 Aquaculture with Laboratory; ZO 515 Fish Physiology), hands on training of postdoctoral, graduate and undergraduate students during execution of the research, and direct contact with Hybrid striped bass growers during research conducted, in part, on their farms.
PROJECT MODIFICATIONS: None to date.
Impacts
Biofiltration: Results from the biofilter studies directly impacted how a large scale fish farm in North Carolina was designed. Similarly, results from this study also impacted the design of biofilters at a large research facility in Florida. Flounder: Results from the studies on flounder sex determination have been used to design the facilities (level of illumination) and modify production practices (age at which fish are exposed to certain environmental cues) to maximize the number of females in a production cohort. These modifications have been applied to a new commercial southern flounder farm currently in its first year of operation. Compensatory Growth: We found that compensatory growth (CG) can be induced in hybrid striped bass (HSB) fed daily after 3-week periods of no feeding. The CG response is characterized by increases in specific growth rate (SGR), hyperphagia and improved feed conversion. Insulin-like growth factor-I (IGF-I) is a critical growth regulator
in fish and the IGF binding proteins (IGFBPs) prolong IGF-I's half-life and regulates its availability to target tissues. During the feed restriction period, circulating insulin-like growth factor-I (IGF-I) and the IGFBP-3 declines in parallel with SGR. Upon refeeding, IGF-I and IGFBP-3 rapidly increase along with SGR. Overall, significant correlations between SGR and plasma IGF-I were seen during periods of slow, normal and accelerated growth in these experiments. Based on this work we conclude that IGFBP-3 may serve to prolong the growth promoting actions of IGF-I during states of rapid growth. We also have established that plasma IGF-I is a good bioindicator of growth in fish, and may therefore prove an effective tool for assessing the growth status of fish in their environment. Egg Quality: Results of the basic studies on oogenesis and egg quality revealed the presence and characteristics of three different forms of Vtg yolk precursors in temperate basses and detailed comparisons
of the primary structures of each Vtg with partial or complete sequences of the bass yolk proteins or of Vtgs from other fishes revealed conserved and variant structural elements of teleost Vtgs with functional significance, including, as examples, dimerization sites, receptor-binding sites, and enzyme recognitions sites. In addition, multiple oocyte receptors for the three forms of Vtg were discovered for the first time in any vertebrate. These finding led to our development of a new model for oocyte growth in teleost fishes and set the stage and capability for our final analyses of the blood and ovarian samples from maturing broodstock. The new model for oocyte growth was explored and found to be applicable in several additional species (grey mullet, Mozambique tilapia, red-tailed goodeid, medaka, barfin flounder and red seabream) in collaborative research involving exchange of ideas and findings with several colleagues in the US and Japan.
Publications
- Picha, M.E., Silverstein, J.T. and Borski, R.J. (2006). Discordant regulation of hepatic IGF-I mRNA and circulating IGF-I during compensatory growth in a teleost, the hybrid striped bass (Morone chrysops X M. saxatilis). General and Comparative Endocrinology, 147:196-205.
- Amano, H., Fujita, T., Hiramatsu, N., Kagawa, H., Sawaguchi, S., Matsubara, T., Sullivan, C.V., and A. Hara. (2007). Molecular alteration of three forms of vitellogenins and their derivative yolk proteins during oocyte growth and maturation in grey mullet (Mugil cephalus). Cybium, the International Journal of Ichthyology (in press).
- Davis, L., Hiramatsu, N., Sullivan, C.V., Hirano, T., and E.G. Grau. (2007). Estrogen regulation of multiple vitellogenin and estrogen receptor genes and of the growth hormone-insulin like growth factor axis in tilapia. Cybium, the International Journal of Ichthyology (in press).
- Hiramatsu, N., Mihoka, I., Ideuchi, H., Fujita, T., Amano, H., Matsubara, T., Sullivan, C.V., and A. Hara. (2007). Differential production and uptake of dual vitellogenins in Japanese medaka (Oryzias latipes). Cybium, the International Journal of Ichthyology (in press).
- Reading, B.J., Hiramatsu, N., Matsubara, T., Hara, A., and C.V. Sullivan. (2007). Deduced primary structures of three vitellogenins and specific binding to multiple ovarian receptors in white perch (Morone americana). Cybium, the International Journal of Ichthyology (in press).
- Sawaguchi, S., Ohkubo, N., Amano, H., Hiramatsu, N., Hara, A., Sullivan, C.V., and T. Matsubara. (2007). Controlled accumulation of multiple vitellogenins into oocytes during vitellogenesis in the barfin flounder, Varasper moseri. Cybium, the International Journal of Ichthyology (in press).
- Amano, H., Fujita, T., Hiramatsu, N., Sawaguchi, S., Matsubara, T., Sullivan, C.V. and A. Hara. (2007). Purification of multiple vitellogenins in grey mullet (Mugil cephalus). Mar. Biol. 152(6):1215-1225.
- Saliling, W.J.B., Westerman, P.W., Losordo, T.M. (2007). Wood chips and wheat straw as alternative biofilter media for denitrification reactors treating aquaculture and other wastewaters with high nitrate concentrations. Aquacultural Engineering 37: 222-233.
- Picha, M.E., Turano, M.J. Beckman, B.R., Borski, R.J. (2008) Hormonal biomarkers of growth and applications to aquaculture: A mini-review of growth hormone, insulin-like growth gactor-I and IGF binding proteins as growth indicators in fish. North Am. J. Aquaculture. In press.
- Davis, L.K., Hiramatsu, N., Hiramatsu, K., Reading, B.J., Matsubara, T., Hara, A., Sullivan, C.V., Pierce, A.L., Hirano, T., and E. Gordon Grau. (2007). Induction of three vitellogenins by 17-estradiol with concurrent inhibition of the growth hormone-insulin-like growth factor 1 axis in a euryhaline teleost, the tilapia (Oreochromis mossambicus). Biology of Reproduction 77(4): 614-625.
- Sawaguchi, S., Kagawa, H., Ohkubo, N., Hiramatsu, N., Sullivan, C.V., and T. Matsubara. (2006). Molecular characterization and alteration of three forms of Vg and their yolk protein products during oocyte growth and maturation in red seabream (Pagrus major), a marine teleost spawning pelagic eggs. Molecular Reproduction and Development 73:719-736.
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