UMBC Department of Marine Biotechnology

Permanent URI for this collection

The Department of Marine Biotechnology (DMB) is partner member of the Institute of Marine and Environmental Technology (IMET), a joint University System of Maryland research institute capitalizing on the strengths of the University of Maryland, Baltimore County, the University of Maryland Center for Environmental Sciences, and the University of Maryland, Baltimore. DMB is premier research center for basic and translational research in marine biotechnology and molecular microbiology. Researchers use the tools of modern molecular biology and biotechnology to make basic scientific discoveries that may be applied to study, protect and enhance marine and estuarine resources. DMB faculty focus on key areas of marine molecular biology to solve practical problems confronting the Chesapeake Bay and the greater marine ecosystem and to develop new technologies for translational application and economic advancement. DMB provides an exceptional environment for specialized training and mentoring of tomorrow’s biotechnology workforce.

The department is based at the Columbus Center in downtown Baltimore. Faculty have access to extensive research facilities, including an 18,000 square-foot, fully-contained, recirculating marine aquaculture facility, a versatile high-volume fermentation facility, and a wide assortment of advanced biomolecular instrumentation.

Browse

Recent Submissions

Now showing 1 - 20 of 62
  • Item
    Identifying the genes involved in the egg-carrying ovigerous hair development of the female blue crab Callinectes sapidus: transcriptomic and genomic expression analyses
    (BMC Genomics, 2023-12-11) Wang, Tao; Bachvaroff, Tsvetan; Chung, J. Sook
    Background Crustacean female sex hormone (CFSH) controls gradually developing adult female-specific morphological features essential for mating and brood care. Specifically, ovigerous hairs are developed during the prepuberty molt cycle of the blue crab Callinectes sapidus that are essential for carrying the eggs until they finish development. Reduced CFSH transcripts by CFSH-dsRNA injections result in fewer and shorter ovigerous hairs than the control. This study aimed to identify the specific genes responsible for ovigerous hair formation using transcriptomic, genomic and expression analyses of the ovigerous setae at three stages: prepuberty at early (OE) and late premolt (OL), and adult (AO) stages. Results The de novo Trinity assembly on filtered sequence reads produced 96,684 Trinity genes and 124,128 transcripts with an N50 of 1,615 bp. About 27.3% of the assembled Trinity genes are annotated to the public protein sequence databases (i.e., NR, Swiss-Prot, COG, KEGG, and GO databases). The OE vs. OL, OL vs. AO, and OE vs. AO comparisons resulted in 6,547, 7,793, and 7,481 differentially expressed genes, respectively, at a log2-fold difference. Specifically, the genes involved in the Wnt signaling and cell cycle pathways are positively associated with ovigerous hair development. Moreover, the transcripts of ten cuticle protein genes containing chitin-binding domains are most significantly changed by transcriptomic analysis and RT-qPCR assays, which shows a molt-stage specific, down-up-down mode across the OE-OL-AO stages. Furthermore, the expression of the cuticle genes with the chitin-binding domain, Rebers and Riddiford domain (RR)-1 appears at early premolt, followed by RR-2 at late premolt stage. Mapping these 10 cuticle protein sequences to the C. sapidus genome reveals that two scaffolds with a 549.5Kb region and 35 with a 1.19 Mb region harbor 21 RR1 and 20 RR2 cuticle protein genes, respectively. With these findings, a putative mode of CFSH action in decapod crustaceans is proposed. Conclusions The present study describes a first step in understanding the mechanism underlying ovigerous hair formation in C. sapidus at the molecular level. Overall, demonstrating the first transcriptome analysis of crustacean ovigerous setae, our results may facilitate future studies into the decapod female reproduction belonging to the suborder Pleocyemata.
  • Item
    Induction of Reproductive Sterility in Coho Salmon (Oncorhynchus kisutch) by an Immersion-Based Gene Silencing Technology
    (MDPI, 2023-11-21) Xu, Lan; Zhao, Mingli; Zohar, Yonathan; Wong, Ten-Tsao
    Undesired maturation and reproduction are major challenges in fish aquaculture regarding genetic introgression, precocious maturation, and reproduction-related mortality, which can have profound ecological or economic impacts. Farming reproductively sterile fish can effectively mitigate these challenges. In this paper, we transferred and applied a novel immersion-based, non-transgenic gene silencing technology to sterilize coho salmon for the first time. Unfertilized eggs were bath immersion-treated with csdnd-MO-Vivo in different immersion media. Eyed rates of treated groups ranged from 0.9 to 63.5%. Sterile fish lacking germ cells, and those with arrested germ cells/atretic oocytes, were obtained at 14 and 20 months of age, albeit at a low percentage (2.3 to 10.0% based on females). Gonadal histology and vasa/nanos3 gene expression profile were provided for comparing fertile and sterile gonads, as well as retarded ovaries. Future directions and strategies for optimizing the technology and improving sterility induction were also proposed. The successful production of sterile coho salmon achieved in this study demonstrates the proof of principle for this new sterilization technology. As we continue to expand upon these findings and refine the technology, achieving coho salmon sterile population farming would facilitate the future transfer and application to other commercially important aquaculture fish.
  • Item
    Oyster reef habitat depends on environmental conditions and management across large spatial scales
    (Inter-Research Science Publisher, 2023-10-19) Tracy, Allison M.; Heggie, Keira; Ritter, Carmen; Norman, Dave; Aguilar, Rob; Ogburn, Matthew B.
    Oyster reefs provide important services to ecosystems and people, with many of these benefits depending on structurally complex reef habitat. Despite the key role of oyster reef habitat, we have yet to understand natural and anthropogenic drivers of subtidal reef habitat over large spatial scales (>200 km). Chesapeake Bay (USA) offers a valuable system to explore how salinity, restoration, and harvest compare in their influence on subtidal oyster reef habitat because of its broad environmental gradient and mosaic of management types. We applied a remote rapid assessment method using underwater photographs to survey oyster reef habitat in 12 tributaries and scored images based on estimates of oyster percent cover and vertical relief. The broad spatial scale (~215 km) of the survey includes reefs that vary in management status and salinity. Bay-wide habitat scores were higher with greater estimated oyster percent cover and vertical relief on unharvested and restored reefs. Salinity also contributed to Chesapeake Bay-wide patterns, but the relationship depended on harvest status. In assessing the separate management jurisdictions, scores were higher on restored reefs in Maryland and on anthropogenic (i.e. artificially supplemented) reefs in Virginia. A time series over 4 yr in 2 Maryland tributaries showed high and persistent habitat scores in restored sanctuaries, but habitat scores increased for all reefs over time. The results highlight the combined roles of the natural environment and management decisions on oyster reef habitat. The effect of harvest and restoration on habitat underscores the importance of local management decisions in determining the future status of oyster reefs.
  • Item
    Cross-Hemispheric Genetic Diversity and Spatial Genetic Structure of Callinectes sapidus Reovirus 1 (CsRV1)
    (MDPI, 2023-02-18) Zhao, Mingli; Plough, Louis V.; Behringer, Donald C.; Bojko, Jamie; Kough, Andrew S.; Alper, Nathaniel W.; Xu, Lan; Schott, Eric J.
    The movement of viruses in aquatic systems is rarely studied over large geographic scales. Oceanic currents, host migration, latitude-based variation in climate, and resulting changes in host life history are all potential drivers of virus connectivity, adaptation, and genetic structure. To expand our understanding of the genetic diversity of Callinectes sapidus reovirus 1 (CsRV1) across a broad spatial and host life history range of its blue crab host (Callinectes sapidus), we obtained 22 complete and 96 partial genomic sequences for CsRV1 strains from the US Atlantic coast, Gulf of Mexico, Caribbean Sea, and the Atlantic coast of South America. Phylogenetic analyses of CsRV1 genomes revealed that virus genotypes were divided into four major genogroups consistent with their host geographic origins. However, some CsRV1 sequences from the US mid-Atlantic shared high genetic similarity with the Gulf of Mexico genotypes, suggesting potential human-mediated movement of CsRV1 between the US mid-Atlantic and Gulf coasts. This study advances our understanding of how climate, coastal geography, host life history, and human activity drive patterns of genetic structure and diversity of viruses in marine animals and contributes to the capacity to infer broadscale host population connectivity in marine ecosystems from virus population genetic data.
  • Item
    Pacific oysters are a sink and a potential source of the eelgrass pathogen, Labyrinthula zosterae
    (Inter-Research, 2022-12-15) Agnew, M Victoria; Groner, Maya L.; Eisenlord, Morgan E.; Friedman, Carolyn S.; Burge, Colleen
    Oyster aquaculture and seagrasses often co-occur and are each vital to the ecological and economic value of coastal ecosystems. Global declines in seagrasses, including Zostera mari - na, have recently been observed in association with multiple factors, including infection with diseases such as seagrass wasting disease (SWD), caused by the protist Labyrinthula zosterae. Protection of seagrasses has led to restrictions on oyster aquaculture due to perceived negative impacts on seagrass beds; however, positive impacts may also occur. An important aquaculture species, the Pacific oyster Crassostrea gigas, can filter L. zosterae from the water, potentially re - ducing pathogen transmission, although oysters may vector infection if they accumulate and re - lease live L. zosterae into the water. We investigated whether oyster presence decreases lesion severity and infection intensity in eelgrass, or acts as a vector of L. zosterae, via laboratory and field experiments. In the laboratory, oysters and eelgrass were exposed to L. zosterae for 24 h and kept at 11°C or 18°C for 13 d. In the field, eelgrass ramets were deployed with and without oysters for 28 d adjacent to eelgrass known to have SWD. In the laboratory experiment, the presence of oysters significantly decreased lesion severity and infection intensity, but oysters previously ex - posed to L. zosterae did transmit the pathogen to naïve eelgrass. Temperature did not affect oyster ability to mitigate SWD; however, increased temperature significantly increased lesion severity. Oysters had no effect on SWD in the field. Further research is needed regarding the potential for oysters to vector L. zosterae and to quantify when oysters reduce SWD in the field.
  • Item
    Seasonal trends of PCBs in air over Washington DC reveal localized urban sources and the influence of Anacostia River
    (Elsevier, 2022-10-27) Bokare, Mandar; Lombard, Nathalie; Magee, Samuel; Murali, Dev; Ghosh, Upal
    Semi-volatile organic compounds like polychlorinated biphenyls (PCBs) undergo diffusive exchange flux between a water body and the overlying air. The magnitude of this exchange can be a substantial component of the overall pollutant mass balance and needs to be determined accurately to identify major pollutant sources to the water body and to plan appropriate remedies. For the PCB-impacted Anacostia River in Washington DC (USA), quantification of air-water exchange has been a major data gap. In the present study, polyethylene passive samplers were used to measure PCB concentrations in air phase at six locations in DC over a period of one year to capture spatial and seasonal variations. Concurrent water phase PCB measurements were used to quantify the direction and magnitude of air-water exchange in the Anacostia River. Two locations had nearly an order of magnitude higher air phase PCB concentrations that could be related to localized sources. Remaining four locations provided similar air phase PCB concentrations that averaged from 270 ± 44 pg/m3 (summer) to 32 ± 4.3 pg/m3 (winter). ∑PCB water-air exchange fluxes were positive across all seasons, with net PCB volatilization of 180 ± 19 g/year from the surface water. Volatilization rate was an order of magnitude lower than previously estimated from a fate and transport model. PCB load from atmospheric deposition based on previous studies in this watershed was an order of magnitude lower than the volatilization rate. Results refuted a long-standing understanding of the air phase serving as a source of PCBs to the river as per the currently approved Total Maximum Daily Load assessment. The study demonstrates the utility of passive air phase measurements in delineating local terrestrial sources of pollution as well as providing estimates for air-water exchange to complete a robust mass balance for semi-volatile pollutants in an urban river.
  • Item
    Genetic compensation of triacylglycerol biosynthesis in the green microalga Chlamydomonas reinhardtii
    (Wiley, 2022-06-21) Lee, Yi-Ying; Park, Rudolph; Miller, Stephen M.; Li, Yantao
    Genetic compensation has been proposed to explain phenotypic differences between gene knockouts and knockdowns in several metazoan and plant model systems. With the rapid development of reverse genetic tools such as CRISPR/Cas9 and RNAi in microalgae, it is increasingly important to assess whether genetic compensation affects the phenotype of engineered algal mutants. While exploring triacylglycerol (TAG) biosynthesis pathways in the model alga Chlamydomonas reinhardtii, it was discovered that knockout of certain genes catalyzing rate-limiting steps of TAG biosynthesis, type-2 diacylglycerol acyltransferase genes (DGTTs), triggered genetic compensation under abiotic stress conditions. Genetic compensation of a DGTT1 null mutation by a related PDAT gene was observed regardless of the strain background or mutagenesis approach, for example, CRISPR/Cas 9 or insertional mutagenesis. However, no compensation was found in the PDAT knockout mutant. The effect of PDAT knockout was evaluated in a Δvtc1 mutant, in which PDAT was upregulated under stress, resulting in a 90% increase in TAG content. Knockout of PDAT in the Δvtc1 background induced a 12.8-fold upregulation of DGTT1 and a 272.3% increase in TAG content in Δvtc1/pdat1 cells, while remaining viable. These data suggest that genetic compensation contributes to the genetic robustness of microalgal TAG biosynthetic pathways, maintaining lipid and redox homeostasis in the knockout mutants under abiotic stress. This work demonstrates examples of genetic compensation in microalgae, implies the physiological relevance of genetic compensation in TAG biosynthesis under stress, and provides guidance for future genetic engineering and mutant characterization efforts.
  • Item
    Reproductive sterility in aquaculture: A review of induction methods and an emerging approach with application to Pacific Northwest finfish species
    (Wiley, 2022-07-20) Xu, Lan; Zhao, Mingli; Ryu, Jun Hyung; Hayman, Edward S.; Fairgrieve, William T.; Zohar, Yonathan; Luckenbach, J. Adam; Wong, Ten-Tsao
    Aquaculture is the fastest-growing food-production sector and is striving to become a long-term sustainable approach to meet the rising global demand for seafood. During the expansion and advancement of aquaculture, minimizing ecological impacts should occur concomitantly with maximizing production. Farmed fish, often genetically distinct from their natural conspecifics, may pose significant risks of genetic contamination and ecological imbalance to wild populations if they escape from aquaculture confinement. Growing reproductively sterile fish is the most effective way to genetically contain farmed fish. Atlantic salmon (Salmo salar) escape events in the ‘Pacific Northwest’ region of the United States and Canada have raised alarms over potential ecological impacts and led to legislation in Washington State phasing out the culture of non-native finfish species. Farming sterile native species such as coho salmon (Oncorhynchus kisutch) and sablefish (Anoplopoma fimbria) in the Pacific Northwest would ease public concerns and promote environmentally and economically sustainable aquaculture. Sterile fish also can mitigate the challenge of precocious maturation, a prominent issue associated with culture of salmonids and many other species, to improve somatic growth, flesh quality and fish health and welfare. Here, we review methods having potential applications for producing sterile fish and introduce our novel immersion-based technology that temporarily silences the dead end (dnd) gene using Morpholino oligonucleotides to produce sterile coho salmon and sablefish for the first time. The successful induction of sterility in these two iconic Pacific Northwest species without introducing genetic modifications would promote the use of this immersion-based sterilization technology for more aquaculture finfish worldwide.
  • Item
    Intranasal vaccination of hamsters with a Newcastle disease virus vector expressing the S1 subunit protects animals against SARS-CoV-2 disease
    (Nature, 2022-06-20) Díaz, Manolo Fernández; Calderón, Katherine; Rojas-Neyra, Aldo; Vakharia, Vikram; et al
    The coronavirus disease-19 (COVID-19) pandemic has already claimed millions of lives and remains one of the major catastrophes in the recorded history. While mitigation and control strategies provide short term solutions, vaccines play critical roles in long term control of the disease. Recent emergence of potentially vaccine-resistant and novel variants necessitated testing and deployment of novel technologies that are safe, effective, stable, easy to administer, and inexpensive to produce. Here we developed three recombinant Newcastle disease virus (rNDV) vectored vaccines and assessed their immunogenicity, safety, and protective efficacy against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in mice and hamsters. Intranasal administration of rNDV-based vaccine candidates elicited high levels of neutralizing antibodies. Importantly, the nasally administrated vaccine prevented lung damage, and significantly reduced viral load in the respiratory tract of vaccinated animal which was compounded by profound humoral immune responses. Taken together, the presented NDV-based vaccine candidates fully protected animals against SARS-CoV-2 challenge and warrants evaluation in a Phase I human clinical trial as a promising tool in the fight against COVID-19.
  • Item
    Genetic subtyping and phylogenetic analysis of HA and NA from avian influenza virus in wild birds from Peru reveals unique features among circulating strains in America
    (PLOS, 2022-06-07) Castro-Sanguinetti, Gina R.; Simas, Paulo Vitor Marques; Apaza-Chiara, Ana Paola; Callupe-Leyva, Jose Alonso; Rondon-Espinoza, Juan Alexander; Gavidia, Cesar M.; More-Bayona, Juan Anderson; Veliz, Rosa Isabel Gonzalez; Vakharia, Vikram; Icochea, Maria Eliana
    Avian influenza virus (AIV) represents a major concern with productive implications in poultry systems but it is also a zoonotic agent that possesses an intrinsic pandemic risk. AIV is an enveloped, negative-sense and single-stranded RNA virus with a segmented genome. The eight genomic segments, comprising the whole genome, encode for eleven proteins. Within these proteins, Hemagglutinin (HA) and Neuraminidase (NA) are the most relevant for studies of evolution and pathogenesis considering their role in viral replication, and have also been used for classification purposes. Migratory birds are the main hosts and play a pivotal role in viral evolution and dissemination due to their migratory routes that comprise large regions worldwide. Altogether, viral and reservoir factors contribute to the emergence of avian influenza viruses with novel features and pathogenic potentials. The study aimed to conduct surveillance of AIVs in wild birds from Peru. A multi-site screening of feces of migratory birds was performed to isolate viruses and to characterize the whole genome sequences, especially the genes coding for HA and NA proteins. Four-hundred-twenty-one (421) fecal samples, collected between March 2019 and March 2020 in Lima, were obtained from 21 species of wild birds. From these, we isolated five AIV from whimbrel, kelp gull, Franklin’s gulls and Mallard, which were of low pathogenicity, including four subtypes as H6N8, H13N6, H6N2 and H2N6. Genetic analysis of HA and NA genes revealed novel features in these viruses and phylogenetic analysis exhibited a close relationship with those identified in North America (US and Canada). Furthermore, H2N6 isolate presented a NA sequence with higher genetic relationship to Chilean isolates. These results highlight that the geographical factor is of major relevance in the evolution of AIV, suggesting that AIV circulating in Peru might represent a new site for the emergence of reassortant AIVs.
  • Item
    Chemogenetic Depletion of Hypophysiotropic GnRH Neurons Does Not Affect Fertility in Mature Female Zebrafish
    (MDPI, 2022-05-17) Tanaka, Sakura; Zmora, Nilli; Levavi-Sivan, Berta; Zohar, Yonathan
    The hypophysiotropic gonadotropin-releasing hormone (GnRH) and its neurons are crucial for vertebrate reproduction, primarily in regulating luteinizing hormone (LH) secretion and ovulation. However, in zebrafish, which lack GnRH1, and instead possess GnRH3 as the hypophysiotropic form, GnRH3 gene knockout did not affect reproduction. However, early-stage ablation of all GnRH3 neurons causes infertility in females, implicating GnRH3 neurons, rather than GnRH3 peptides in female reproduction. To determine the role of GnRH3 neurons in the reproduction of adult females, a Tg(gnrh3:Gal4ff; UAS:nfsb-mCherry) line was generated to facilitate a chemogenetic conditional ablation of GnRH3 neurons. Following ablation, there was a reduction of preoptic area GnRH3 neurons by an average of 85.3%, which was associated with reduced pituitary projections and gnrh3 mRNA levels. However, plasma LH levels were unaffected, and the ablated females displayed normal reproductive capacity. There was no correlation between the number of remaining GnRH3 neurons and reproductive performance. Though it is possible that the few remaining GnRH3 neurons can still induce an LH surge, our findings are consistent with the idea that GnRH and its neurons are likely dispensable for LH surge in zebrafish. Altogether, our results resurrected questions regarding the functional homology of the hypophysiotropic GnRH1 and GnRH3 in controlling ovulation.
  • Item
    Current advances in molecular, biochemical, and computational modeling analysis of microalgal triacylglycerol biosynthesis
    (Elsevier, 2016-07-14) Lenka, Sangram; Carbonaro, Nicole; Park, Rudolph; Miller, Stephen M.; Thorpe, Ian; Li, Yantao
    Triacylglycerols (TAGs) are highly reduced energy storage molecules ideal for biodiesel production. Microalgal TAG biosynthesis has been studied extensively in recent years, both at the molecular level and systems level through experimental studies and computational modeling. However, discussions of the strategies and products of the experimental and modeling approaches are rarely integrated and summarized together in a way that promotes collaboration among modelers and biologists in this field. In this review, we outline advances toward understanding the cellular and molecular factors regulating TAG biosynthesis in unicellular microalgae with an emphasis on recent studies on rate-limiting steps in fatty acid and TAG synthesis, while also highlighting new insights obtained from the integration of multi-omics datasets with mathematical models. Computational methodologies such as kinetic modeling, metabolic flux analysis, and new variants of flux balance analysis are explained in detail. We discuss how these methods have been used to simulate algae growth and lipid metabolism in response to changing culture conditions and how they have been used in conjunction with experimental validations. Since emerging evidence indicates that TAG synthesis in microalgae operates through coordinated crosstalk between multiple pathways in diverse subcellular destinations including the endoplasmic reticulum and plastids, we discuss new experimental studies and models that incorporate these findings for discovering key regulatory checkpoints. Finally, we describe tools for genetic manipulation of microalgae and their potential for future rational algal strain design. This comprehensive review explores the potential synergistic impact of pathway analysis, computational approaches, and molecular genetic manipulation strategies on improving TAG production in microalgae.
  • Item
    Advantages, Factors, Obstacles, Potential Solutions, and Recent Advances of Fish Germ Cell Transplantation for Aquaculture—A Practical Review
    (MDPI, 2022-02-10) Ryu, Jun Hyung; Xu, Lan; Wong, Ten-Tsao
    Germ cell transplantation technology enables surrogate offspring production in fish. This technology has been expected to mitigate reproductive barriers, such as long generation time, limited fecundity, and complex broodstock management, enhancing seed production and productivity in aquaculture. Many studies of germ cell transplantation in various fish species have been reported over a few decades. So far, surrogate offspring production has been achieved in many commercial species. In addition, the knowledge of fish germ cell biology and the related technologies that can enhance transplantation efficiency and productivity has been developed. Nevertheless, the commercial application of this technology still seems to lag behind, indicating that the established models are neither beneficial nor cost-effective enough to attract potential commercial users of this technology. Furthermore, there are existing bottlenecks in practical aspects such as impractical shortening of generation time, shortage of donor cells with limited resources, low efficiency, and unsuccessful surrogate offspring production in some fish species. These obstacles need to be overcome through further technology developments. Thus, we thoroughly reviewed the studies on fish germ cell transplantation reported to date, focusing on the practicality, and proposed potential solutions and future perspectives.
  • Item
    UNDER THE BAY: An Augmented Reality Project
    (2021) Bachvaroff, Tsvetan; Moren, Lisa; Deacon, Dan; Olano, Marc
    “Under the Bay” tells the story of a world beneath the surface of the Chesapeake Bay and really all estuaries and oceans. In this augmented reality project a user can point their cell phone at the water — or anywhere in the world — like a microscope and see and hear the hidden invisibilities under the Chesapeake Bay. When they do, a series of stories between humans and non-humans emerge told by artist Lisa Moren with music by electronic composer Dan Deacon. Directed by Lisa Moren and marine biologist Tsvetan Bachvaroff, the team worked with software engineer Marc Olano to develop the project.
  • Item
    Vasoactive intestinal peptide indirectly elicits pituitary LH secretion independent of GnRH in female zebrafish
    (Oxford University Press, 2022-01-03) Tanaka, Sakura; Zmora, Nilli; Levavi-Sivan, Berta; Zohar, Yonathan
    Vasoactive intestinal peptide (Vip) regulates luteinizing hormone (LH) release through the direct regulation of gonadotropin-releasing hormone (GnRH) neurons at the level of the brain in female rodents. However, little is known regarding the roles of Vip in teleost reproduction. Although GnRH is critical for fertility through the regulation of LH secretion in vertebrates, the exact role of the hypophysiotropic GnRH (GnRH3) in zebrafish is unclear since GnRH3 null fish are reproductively fertile. This phenomenon raises the possibility of a redundant regulatory pathway(s) for LH secretion in zebrafish. Here, we demonstrate that VipA (homologues of mammalian Vip) both inhibits and induces LH secretion in zebrafish. Despite the observation that VipA axons may reach the pituitary proximal pars distalis including LH cells, pituitary incubation with VipA in vitro, and intraperitoneal injection of VipA, did not induce LH secretion and lhβ mRNA expression in sexually mature females, respectively. On the other hand, intracerebroventricular administration of VipA augmented plasma LH levels in both wild type and gnrh3-/- females at 1 hour post-treatment, with no observed changes in pituitary GnRH2 and GnRH3 contents and gnrh3 mRNA levels in the brains. While VipA’s manner of inhibition of LH secretion has yet to be explored, the stimulation seems to occur via a different pathway than GnRH3, dopamine, and E2 in regulating LH secretion. The results indicate that VipA induces LH release possibly by acting with or through a non-GnRH factor(s), providing proof for the existence of functional redundancy of LH release in sexually mature female zebrafish.
  • Item
    Newcastle Disease Virus Vectored Chicken Infectious Anaemia Vaccine Induces Robust Immune Response in Chickens
    (MDPI, 2021-10-02) Chellappa, Madhan Mohan; Dey, Sohini; Pathak, Dinesh Chandra; Singh, Asmita; Ramamurthy, Narayan; Ramakrishnan, Saravanan; Mariappan, Asok Kumar; Dhama, Kuldeep; Vakharia, Vikram
    Newcastle disease virus (NDV) strain R2B, with an altered fusion protein cleavage site, was used as a viral vector to deliver the immunogenic genes VP2 and VP1 of chicken infectious anaemia virus (CIAV) to generate a bivalent vaccine candidate against these diseases in chickens. The immunogenic genes of CIAV were expressed as a single transcriptional unit from the NDV backbone and the two CIA viral proteins were obtained as separate entities using a self-cleaving foot-and-mouth disease virus 2A protease sequence between them. The recombinant virus (rR2B-FPCS-CAV) had similar growth kinetics as that of the parent recombinant virus (rR2B-FPCS) in vitro with similar pathogenicity characteristics. The bivalent vaccine candidate when given in specific pathogen-free chickens as primary and booster doses was able to elicit robust humoral and cell-mediated immune (CMI) responses obtained in a vaccination study that was conducted over a period of 15 weeks. In an NDV and CIAV ELISA trial, there was a significant difference in the titres of antibody between vaccinated and control groups which showed slight reduction in antibody titre by 56 days of age. Hence, a second booster was administered and the antibody titres were maintained until 84 days of age. Similar trends were noticed in CMI response carried out by lymphocyte transformation test, CD4+ and CD8+ response by flow cytometry analysis and response of real time PCR analysis of cytokine genes. Birds were challenged with virulent NDV and CIAV at 84 days and there was significant reduction in the NDV shed on the 2nd and 4th days post challenge in vaccinated birds as compared to unvaccinated controls. Haematological parameters comprising PCV, TLC, PLC and PHC were estimated in birds that were challenged with CIAV that indicated a significant reduction in the blood parameters of controls. Our findings support the development and assessment of a bivalent vaccine candidate against NDV and CIAV in chickens.
  • Item
    First data on aquaculture of the Tripletail, Lobotes surinamensis, a promising candidate species for U.S. marine aquaculture
    (Wiley Online Library, 2021-05-18) Saillant, Eric; Adams, Nicholas; Lemus, Jason T.; Franks, James S.; Zohar, Yonathan; Stubblefield, John; Manley, Christopher
    The Tripletail, Lobotes surinamensis, is a warm-water pelagic fish that is increasingly targeted by U.S. anglers. The superior quality of Tripletail flesh coupled with the lack of domestic commercial fisheries stimulated interests to develop aquaculture of this species. In this work, photo-thermal conditioning of captive-held broodstocks promoted maturation in females, but spontaneous spawning was not observed. GnRHa slow-release implants induced ovulation in late vitellogenic females but fertility remained below 10% when GnRHa was administered alone. However, spawns with high fertility (up to 85%) were obtained when a dopamine antagonist was administered in conjunction with GnRHa implants indicating dopamine inhibition impaired final gamete maturation, in particular sperm production in males, in aquaculture conditions. Tripletail larvae successfully initiated exogenous feeding on enriched rotifers followed by Artemia nauplii and were weaned to prepared feeds at 25 days post hatch, yet with low survival through the late phases of larval culture. Pilot grow-out trials at low density in recirculating systems revealed impressive growth rates averaging over 170 g/month through a market size above 1 kg. While protocols for hatchery culture and grow-out still need to be optimized, current data suggest that Tripletail could become a successful species for U.S. marine aquaculture.
  • Item
    Whole transcriptome analysis reveals changes in expression of immune-related genes during and after bleaching in a reef-building coral
    (The Royal Society, 2015-04-01) Pinzón, Jorge H.; Kamel, Bishoy; Burge, Colleen; Harvell, C. Drew; Medina, Mónica; Weil, Ernesto; Mydlarz, Laura D.
    Climate change is negatively affecting the stability of natural ecosystems, especially coral reefs. The dissociation of the symbiosis between reef-building corals and their algal symbiont, or coral bleaching, has been linked to increased sea surface temperatures. Coral bleaching has significant impacts on corals, including an increase in disease outbreaks that can permanently change the entire reef ecosystem. Yet, little is known about the impacts of coral bleaching on the coral immune system. In this study, whole transcriptome analysis of the coral holobiont and each of the associate components (i.e. coral host, algal symbiont and other associated microorganisms) was used to determine changes in gene expression in corals affected by a natural bleaching event as well as during the recovery phase. The main findings include evidence that the coral holobiont and the coral host have different responses to bleaching, and the host immune system appears suppressed even a year after a bleaching event. These results support the hypothesis that coral bleaching changes the expression of innate immune genes of corals, and these effects can last even after recovery of symbiont populations. Research on the role of immunity on coral's resistance to stressors can help make informed predictions on the future of corals and coral reefs.
  • Item
    Isolation, Identification, and Genomic Analysis of a Novel Reovirus from Healthy Grass Carp and Its Dynamic Proliferation In Vitro and In Vivo
    (MDPI, 2021-04-16) Zhang, Ke; Liu, Wenzhi; Li, Yiqun; Zhou, Yong; Meng, Yan; Zeng, Lingbing; Vakharia, Vikram; Fan, Yuding
    A new grass carp reovirus (GCRV), healthy grass carp reovirus (HGCRV), was isolated from grass carp in 2019. Its complete genome sequence was determined and contained 11 dsRNAs with a total size of 23,688 bp and 57.2 mol% G+C content, encoding 12 proteins. All segments had conserved 5' and 3' termini. Sequence comparisons showed that HGCRV was closely related to GCRV-873 (GCRV-I; 69.57–96.71% protein sequence identity) but shared only 22.65–45.85% and 23.37–43.39% identities with GCRV-HZ08 and Hubei grass carp disease reovirus (HGDRV), respectively. RNA-dependent RNA-polymerase (RdRp) protein-based phylogenetic analysis showed that HGCRV clustered with Aquareovirus-C (AqRV-C) prior to joining a branch common with other aquareoviruses. Further analysis using VP6 amino acid sequences from Chinese GCRV strains showed that HGCRV was in the same evolutionary cluster as GCRV-I. Thus, HGCRV could be a new GCRV isolate of GCRV-I but is distantly related to other known GCRVs. Grass carp infected with HGCRV did not exhibit signs of hemorrhage. Interestingly, the isolate induced a typical cytopathic effect in fish cell lines, such as infected cell shrank, apoptosis, and plague-like syncytia. Further analysis showed that HGCRV could proliferate in grass carp liver (L28824), gibel carp brain (GiCB), and other fish cell lines, reaching a titer of up to 7.5 × 10⁴ copies/μL.