Multifunctional carbonized nanogels to treat lethal acute hepatopancreatic necrosis disease

Abstract Background Shrimp aquaculture has suffered huge economic losses over the past decade due to the outbreak of acute hepatopancreatic necrosis disease (AHPND), which is mainly caused by the bacteria Vibrio parahaemolyticus (V. parahaemolyticus) with the virulence pVA1 plasmid, which encodes a secretory photorhabdus insect-related (Pir) toxin composed of PirA and PirB proteins. The Pir toxin mainly attacks the hepatopancreas, a major metabolic organ in shrimp, thereby causing necrosis and loss of function. The pandemic of antibiotic-resistant strains makes the impact worse. Methods Mild pyrolysis of a mixture of polysaccharide dextran 70 and the crosslinker 1,8-diaminooctane at 180 ℃ for 3 h to form carbonized nanogels (DAO/DEX-CNGs) through controlled cross-linking and carbonization. The multifunctional therapeutic CNGs inherit nanogel-like structures and functional groups from their precursor molecules. Results DAO/DEX-CNGs manifest broad-spectrum antibacterial activity against Vibrio parahaemolyticus responsible for AHPND and even multiple drug-resistant strains. The polymer-like structures and functional groups on graphitic-carbon within the CNGs exhibit multiple treatment effects, including disruption of bacterial membranes, elevating bacterial oxidative stress, and neutralization of PirAB toxins. The inhibition of Vibrio in the midgut of infected shrimp, protection of hepatopancreas tissue from Pir toxin, and suppressing overstimulation of the immune system in severe V. parahaemolyticus infection, revealing that CNGs can effectively guard shrimp from Vibrio invasion. Moreover, shrimps fed with DAO/DEX-CNGs were carefully examined, such as the expression of the immune-related genes, hepatopancreas biopsy, and intestinal microbiota. Few adverse effects on shrimps were observed. Conclusion Our work proposes brand-new applications of multifunctional carbon-based nanomaterials as efficient anti-Vibrio agents in the aquatic industry that hold great potential as feed additives to reduce antibiotic overuse in aquaculture. Graphical Abstract

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Multifunctional Carbonized Nanogels to Treat Lethal Acute Hepatopancreatic Necrosis Disease

10.21203/rs.3.rs-942245/v1 ◽

2021 ◽

Author(s):

Shao-Chieh Yen ◽

Ju-Yi Mao ◽

Hung-Yun Lin ◽

Huai-Ting Huang ◽

Scott G. Harroun ◽

...

Keyword(s):

Functional Groups ◽

Vibrio Parahaemolyticus ◽

Feed Additives ◽

Economic Losses ◽

Multiple Drug ◽

Loss Of Function ◽

Bacterial Membranes ◽

Resistant Strains ◽

Acute Hepatopancreatic Necrosis Disease ◽

The Impact

Abstract Background: Shrimp aquaculture has suffered huge economic losses over the past decade due to the outbreak of acute hepatopancreatic necrosis disease (AHPND), which is mainly caused by the bacteria Vibrio parahaemolyticus (V. parahaemolyticus) with the virulence pVA1 plasmid, which encodes a secretory photorhabdus insect-related (Pir) toxin composed of PirA and PirB proteins. The Pir toxin mainly attacks the hepatopancreas, a major metabolic organ in shrimp, thereby causing necrosis and loss of function. The pandemic of antibiotic-resistant strains makes the impact worse. Methods: Mild pyrolysis of a mixture of polysaccharide dextran 70 and the crosslinker 1,8-diaminooctane at 180 ℃ for 3 h to form carbonized nanogels (DAO/DEX-CNGs) through controlled cross-linking and carbonization. The multifunctional therapeutic CNGs inherit nanogel-like structures and functional groups from their precursor molecules. Results: DAO/DEX-CNGs manifest broad-spectrum antibacterial activity against Vibrio parahaemolyticus responsible for AHPND and even multiple drug-resistant strains. The polymer-like structures and functional groups on graphitic-carbon within the CNGs exhibit multiple treatment effects, including disruption of bacterial membranes, elevating bacterial oxidative stress, and neuralization of PirAB toxins. The inhibition of Vibrio in the midgut of infected shrimp, protection of hepatopancreas tissue from Pir toxin, and suppressing overstimulation of the immune system in severe V. parahaemolyticus infection, revealing that CNGs can effectively guard shrimp from Vibrio invasion. Moreover, shrimps fed with DAO/DEX-CNGs were carefully examined, such as the expression of the immune-related genes, hepatopancreas biopsy, and intestinal microbiota. Few adverse effects on shrimps were observed. Conclusion: Our work proposes brand-new applications of multifunctional carbon-based nanomaterials as efficient anti-Vibrio agents in the aquatic industry that hold great potential as feed additives to reduce antibiotic overuse in aquaculture.

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The infection of Vibrio parahaemolyticus in shrimp and human

Oceana Biomedicina Journal ◽

10.30649/obj.v1i1.6 ◽

2018 ◽

Vol 1 (1) ◽

pp. 44 ◽

Cited By ~ 1

Author(s):

Rian Ka Praja

Keyword(s):

Vibrio Parahaemolyticus ◽

Animal Health ◽

Alternative Therapy ◽

Penaeus Monodon ◽

Early Mortality ◽

Economic Losses ◽

Early Mortality Syndrome ◽

Thermostable Direct Hemolysin ◽

Acute Hepatopancreatic Necrosis Disease ◽

Very High

Vibrio parahaemolyticus is an aquatic zoonotic agent that can threaten human and aquaculture animal health. Humans can be infected by consuming contaminated raw seafood or wound-related infections. Generally infection of V. parahemolyticus is orally transmitted and causes gastroenteritis in humans while in aquaculture animals especially shrimp can cause Acute Hepatopancreatic Necrosis Disease (AHPND) or Early Mortality Syndrome (EMS) with a very high mortality rate and cause economic losses. Shrimp species susceptible to infection are Litopenaeus vannamei, Penaeus monodon, and P. chinensis. V. parahaemolyticus produces several toxins in human disease such as thermostable direct hemolysin (TDH), TDH-related haemolysin (TRH), and thermolabile hemolysin (TLH). Meanwhile, Photorabdus insect-related (Pir) toxins consisting of PirAvp and PirBvp are the toxins associated with AHPND in shrimp. The genes that encode the toxin are used as targets to diagnose V. parahaemolyticus pathogens molecularly. Until now the treatment of V. parahaemolyticus infection is using antibiotics and fluid therapy, but there were V. parahaemolyticus isolates from aquaculture that have been resistant to antibiotics so that the use of antibiotics in aquaculture must be controlled and the use of alternative therapy are very important to be developed to control V. parahaemolyticus infection. Keywords: V. parahaemolyticus, zoonotic, gastroenteritis, Acute Hepatopancreatic Necrosis Disease (AHPND), Early Mortality Syndrome (EMS).

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Molecular characterization of acute hepatopancreatic necrosis disease causing Vibrio parahaemolyticus strains in cultured shrimp Penaeus monodon in south‐west farming region of Bangladesh

Dhaka University Journal of Biological Sciences ◽

10.3329/dujbs.v27i1.46411 ◽

2018 ◽

Vol 27 (1) ◽

pp. 57-68 ◽

Cited By ~ 1

Author(s):

Md Mostavi Enan Eshik ◽

Nusrat Jahan Punom ◽

Mst Khadiza Begum ◽

Tahsin Khan ◽

Mihir Lal Saha ◽

...

Keyword(s):

Vibrio Parahaemolyticus ◽

Penaeus Monodon ◽

16S Rrna Gene Sequencing ◽

Molecular Techniques ◽

Shrimp Farming ◽

Economic Losses ◽

Rrna Gene ◽

South West ◽

Acute Hepatopancreatic Necrosis Disease ◽

Positive Results

Acute hepatopancreatic necrosis disease (AHPND) is an emerging shrimp disease caused by strains of Vibrio parahaemolyticus containing a unique virulent plasmid, responsible for substantial economic losses since 2009; caused up to 100% mortality in farmed shrimp Penaeus monodon. The purpose of this study was to isolate and identify the pathogenic strain of V. parahaemolyticus causing AHPND in cultured shrimp (Penaeus monodon) using classical and molecular techniques. Samples were collected from three different locations of south-west shrimp farming regions of Bangladesh viz. Sadar Upazilla of Satkhira; Mongla and Morrelganj under Bagerhat district. In this study, three selective media were used for primary isolation of V. parahaemolyticus. Among 46 primary isolates, 18 representative isolates were checked for the species-specific detection of V. parahaemolyticus using ldh primers and all of them were found to be positive. 16S rRNA gene sequencing were used to further confirm the isolates as V. parahaemolyticus. tdh primer was used to check human pathogenicity but all 18 isolates showed negative result. The isolates were further characterized to check their AHPND positivity using AP3 and AP4 primers. Ten isolates showed positive results for AP3 (55.56%) and 9 showed positive results for AP4 (50%) which indicated that the isolates were AHPND positive. This study also reported that all AHPND positive strains were resistant to the antibiotic gentamycin but sensitive to chloramphenicol, nalidixic acid, nitrofurantoin and tetracycline. The findings of this study will help the shrimp farmers and policy makers to take proper biosecurity measures to protect shrimps from AHPND and thereby sustain the shrimp production in Bangladesh. Dhaka Univ. J. Biol. Sci. 27(1): 57-68, 2018 (January)

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The impact of catecholamine sensing on the virulence of Vibrio parahaemolyticus causing acute hepatopancreatic necrosis disease (AHPND)

Aquaculture ◽

10.1016/j.aquaculture.2016.12.030 ◽

2017 ◽

Vol 470 ◽

pp. 190-195 ◽

Cited By ~ 7

Author(s):

Nguyen Thao Suong ◽

Nguyen Van Hao ◽

Nguyen Van Sang ◽

Nguyen Dinh Hung ◽

Nguyen Thi Ngoc Tinh ◽

...

Keyword(s):

Vibrio Parahaemolyticus ◽

Acute Hepatopancreatic Necrosis Disease ◽

The Impact

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The Effect of Antibiotic Growth Promoters (AGP) on Antibiotic Resistance and the Digestive System of Broiler Chicken in Sleman, Yogyakarta

BIO Web of Conferences ◽

10.1051/bioconf/20213304005 ◽

2021 ◽

Vol 33 ◽

pp. 04005

Author(s):

Tri Untari ◽

Okti Herawati ◽

Marla Anggita ◽

Widya Asmara ◽

Agnesia Endang Tri Hastuti Wahyuni ◽

...

Keyword(s):

Antibiotic Resistance ◽

Digestive Tract ◽

Broiler Chickens ◽

Broiler Chicken ◽

Feed Additives ◽

Public Understanding ◽

Economic Losses ◽

Growth Promoters ◽

Antibiotic Growth Promoters ◽

The Impact

Chicken is a major source of animal protein consumption in Indonesia. The problem facing the poultry industry is the incidence of resistance which increases mortality of the chicken production. One of the causes of resistance case is the use of antibiotics in feed additives. The public understanding about the effects of the use of antibiotic growth promoters (AGP) in chickens in antibiotic resistance and the digestive tract of chicken needs to be done to avoid the impact on economic losses and health problems. This study aims to provide an understanding of the effects of the use of antibiotic growth promoters (AGP) on antibiotic resistance and the digestive tract of broiler chickens. This study was carried out at a broiler chicken farm in Sleman, Yogyakarta. Based on the histopathological result of the digestive tract of chickens that were given antibiotics as AGP, there was no inflammation occurs, but the administration of antibiotics caused antibiotic resistance in various type of antibiotics including tetracycline (90% resistance), streptomycin (60% resistance), amoxicillin (50% resistance), erythromycin (80% resistance), and no resistance for gentamycin.

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QseC Inhibition as a Novel Antivirulence Strategy for the Prevention of Acute Hepatopancreatic Necrosis Disease (AHPND)-Causing Vibrio parahaemolyticus

Frontiers in Cellular and Infection Microbiology ◽

10.3389/fcimb.2020.594652 ◽

2021 ◽

Vol 10 ◽

Author(s):

Qian Yang ◽

Peizhuo Zou ◽

Zhi Cao ◽

Qingyao Wang ◽

Songzhe Fu ◽

...

Keyword(s):

Broad Spectrum ◽

Vibrio Parahaemolyticus ◽

Pathogenic Bacteria ◽

White Shrimp ◽

Economic Losses ◽

Resistant Bacteria ◽

Flagellar Motility ◽

Type Vi Secretion System ◽

Global Regulation ◽

Acute Hepatopancreatic Necrosis Disease

Acute hepatopancreatic necrosis disease (AHPND) caused by Vibrio parahaemolyticus resulted in great economic losses in global shrimp aquaculture. There is an urgent need for development of novel strategies to combat AHPND-causing V. parahaemolyticus (VpAHPND), given that one of the greatest challenges currently is the widespread use of antibiotics and subsequent emergence of multidrug-resistant bacteria. Here, we proposed a broad-spectrum antivirulence approach targeting a conserved histidine kinase, QseC, which has been demonstrated to activate virulence expression in several Gram-negative pathogens. Our results showed that QseC mediated the catecholamine stimulated effects on growth and flagellar motility of VpAHPND. Transcriptome analysis revealed that QseC was involved in the global regulation of the virulence of VpAHPND as the ΔqseC mutant exhibited a decreased expression of genes related to type IV pilin, flagellar motility, and biofilm formation, while an overexpression of type VI secretion system and cell wall biosynthesis. Subsequently, the bacterial catecholamine receptor antagonist LED209 not only neutralized the stimulatory effects of host catecholamines on the growth and motility of VpAHPNDin vitro, but also attenuated the virulence of VpAHPND towards brine shrimp larvae and white shrimp in vivo. Additionally, LED209 presented no interference with pathogen growth, nor the toxicity to the experimental animals. These results suggest that QseC can be an attractive antivirulence therapy target, and LED209 is a promising candidate for development of broad-spectrum antivirulence agents. This is the first study that demonstrated the role of QseC in the global regulation of VpAHPND infection and demonstrated the antivirulence potential of LED209, which provides insight into the use of an antivirulence approach for targeting not only VpAHPND, but also a much larger collection of pathogenic bacteria.

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Cloning and Expression of PirA gene of Vibrio parahaemolyticus Strain K5 Causing Acute Hepatopancreatic Necrosis Disease in Whiteleg Shrimp in E. coli Host Cell

10.20944/preprints202201.0086.v1 ◽

2022 ◽

Author(s):

Nguyen Quang Linh ◽

Khanh Van Nguyen ◽

Dung Quoc Tran ◽

Van Khanh Tran Quang

Keyword(s):

Recombinant Protein ◽

Vibrio Parahaemolyticus ◽

Expression Vector ◽

Cloning And Expression ◽

Economic Losses ◽

Bacterial Disease ◽

Parahaemolyticus Strain ◽

E Coli ◽

Whiteleg Shrimp ◽

Acute Hepatopancreatic Necrosis Disease

Background: Acute hepatopancreatic necrosis disease (AHPND), is a bacterial disease of whiteleg shrimp, which has a high mortality rate (100%) and incurs economic losses. Our objective was to identify the genes which lead to cell and organ damage and investigate bioproducts to prevent and treat. Methods: Litopenaeus vannamei shrimp in Thua Thien Hue province, Vietnam were collected from an infected pond and analysed at the Institute of Biotechnology, Hue University. The PirA gene of Vibrio parahaemolyticus strain K5 was isolated and analyzed for nucleotide sequence and paired with the expression vector pQE30. The expression vector was transformed into E. coli strain M15, the PirA recombinant protein was expressed in the form of 6xHis-PirA fusion protein of about 15 kDa. PirA recombinant protein was purified and determined the PirAvp binding ratio, cloning and sequencing of PirA gene from Vibrio parahaemolyticus strain K5 causing AHPND by PCR method with specific primers and molecular weights of PirAvp and the PirAvp complex. Results: PirA gene from Vibrio parahaemolyticus strain K5 was cloned into pGEM-T easy vector (Promega, USA) and screened E. coli TOP10 colonies containing pGEM T easy/PirA recombinant plasmid on LB agar/ampicillin/IPTG/X-Gal medium. PCR showing a band of about 347 bp, matching the size of PirA gene and two nucleotide sequences (BamHI and HindIII). The results showed that PirA gene has a length of 336 bp and similar to PirA gene on GenBank (Code: KU556825.1). The results of protein extracted from E. coli M15 recombinant cells and 6xHis-PirA target protein was collected in elution fractions from EF2 to EF6, showed that the concentration of 6xHis-PirA protein and EF3 elution fraction collected a highest protein concentration (1,586.54 µg/ml). Conclusions: The purified PirA recombinant protein will provide materials for development research to create biological products to prevent and treat AHPND.

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Metabolic Alterations in Shrimp Stomach During Acute Hepatopancreatic Necrosis Disease and Effects of Taurocholate on Vibrio parahaemolyticus

Frontiers in Microbiology ◽

10.3389/fmicb.2021.631468 ◽

2021 ◽

Vol 12 ◽

Author(s):

Ramya Kumar ◽

Teng-Chun Tung ◽

Tze Hann Ng ◽

Che-Chih Chang ◽

Yi-Lun Chen ◽

...

Keyword(s):

Bile Acid ◽

Biofilm Formation ◽

Vibrio Parahaemolyticus ◽

Economic Losses ◽

Bacterial Cells ◽

High Concentration ◽

Metabolic Alterations ◽

Biofilm Detachment ◽

Acute Hepatopancreatic Necrosis Disease ◽

Primary Bile Acid

Acute hepatopancreatic necrosis disease (AHPND), a recently emerged bacterial shrimp disease, has increased shrimp mortality and caused huge economic losses in many Asian countries. However, molecular factors underlying pathogenesis of this disease remain largely unknown. Our objective was to characterize metabolic alterations in shrimp stomach during AHPND and determine effects of taurocholate on AHPND-causing Vibrio parahaemolyticus. Based on metabolomics, pathways for lipid metabolism and for primary bile acid (BA) synthesis were majorly affected following AHPND infection. Bile acid metabolites, namely taurocholate, were downregulated in the metabolomics database. This prompted us to study effects of taurocholate on biofilm formation, PirABvp toxin release and biofilm detachment capabilities in AHPND-causing V. parahaemolyticus. Treatment of this bacterium with high concentration of taurocholate, a primary bile acid, induced biofilm formation, PirABvp toxin release and facilitated the dispersion of bacterial cells. Taken together, our findings suggest that AHPND infection can affect the lipid metabolites in shrimp stomach, and further suggest that the primary bile acid taurocholate is important for the virulence of AHPND-causing V. parahaemolyticus.

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THE MAIN DIRECTIONS OF ECONOMIC SUPPORT OF THE REGIONAL SECURITY SYSTEM OF THE POPULATION

EKONOMIKA I UPRAVLENIE: PROBLEMY, RESHENIYA ◽

10.36871/ek.up.p.r.2020.11.01.018 ◽

2020 ◽

Vol 1 (11) ◽

pp. 133-140

Author(s):

E. V. DMITRIEVA ◽

Keyword(s):

Crisis Management ◽

Security System ◽

Regional Security ◽

Economic Losses ◽

Economic Damage ◽

Economic Activities ◽

The Sustainable Development ◽

Economic Support ◽

The Impact ◽

Crisis Situations

The article considers topical issues of economic support for the development of the regional security system of the population against various risks. The dependence of the impact of the scale of crisis situations on economic activities in the constituent entities of the Russian Federation, which become a serious barrier to the sustainable development of the regions of the country, was investigated. The increasing importance of risks of economic losses from accidents and disasters at potentially dangerous facilities as a result of the complex influence of natural, manmade and fire factors has been established. An analysis was carried out and proposals were developed to implement the key tasks of the state in the field of ensuring the protection of the population and territories of the country from disasters in order to ensure the stability of the economy. The organizational structure, division of tasks and functions between officials, crisis management structures and responding units were analyzed, taking into account the reduction in current financial costs. On the basis of a study of the peculiarities of the regions of the country, recommendations were formed to fulfill the necessary tasks by the anti-crisis management bodies in the field of reducing economic damage on the basis of preventing crisis situations and ensuring fire safety. It is proposed to organize the practical application of a complex automated security system based on modern developments with the application of improving the qualities and efficiency of anti-crisis management processes in order to increase economic efficiency. Initial data were formed to reduce potential threats of a natural, man-made, fire and other nature in the regions using financial and economic mechanisms. It is proposed to implement a set of priority measures to further improve and increase the potential of economic support for the anti-crisis management system. The materials of the article can be used in planning the main directions of the development of the regional population security system and the implementation of socio-economic development programs.

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ATP13A2 Regulates Cellular α-Synuclein Multimerization, Membrane Association, and Externalization

International Journal of Molecular Sciences ◽

10.3390/ijms22052689 ◽

2021 ◽

Vol 22 (5) ◽

pp. 2689

Author(s):

Jianmin Si ◽

Chris Van den Haute ◽

Evy Lobbestael ◽

Shaun Martin ◽

Sarah van Veen ◽

...

Keyword(s):

Membrane Integrity ◽

Ubiquitin Proteasome System ◽

Regulatory Function ◽

Membrane Association ◽

Loss Of Function ◽

Atypical Form ◽

Polyamine Transport ◽

Independent Functions ◽

Ubiquitin Proteasome ◽

The Impact

ATP13A2, a late endo-/lysosomal polyamine transporter, is implicated in a variety of neurodegenerative diseases, including Parkinson’s disease and Kufor–Rakeb syndrome, an early-onset atypical form of parkinsonism. Loss-of-function mutations in ATP13A2 result in lysosomal deficiency as a consequence of impaired lysosomal export of the polyamines spermine/spermidine. Furthermore, accumulating evidence suggests the involvement of ATP13A2 in regulating the fate of α-synuclein, such as cytoplasmic accumulation and external release. However, no consensus has yet been reached on the mechanisms underlying these effects. Here, we aimed to gain more insight into how ATP13A2 is linked to α-synuclein biology in cell models with modified ATP13A2 activity. We found that loss of ATP13A2 impairs lysosomal membrane integrity and induces α-synuclein multimerization at the membrane, which is enhanced in conditions of oxidative stress or exposure to spermine. In contrast, overexpression of ATP13A2 wildtype (WT) had a protective effect on α-synuclein multimerization, which corresponded with reduced αsyn membrane association and stimulation of the ubiquitin-proteasome system. We also found that ATP13A2 promoted the secretion of α-synuclein through nanovesicles. Interestingly, the catalytically inactive ATP13A2 D508N mutant also affected polyubiquitination and externalization of α-synuclein multimers, suggesting a regulatory function independent of the ATPase and transport activity. In conclusion, our study demonstrates the impact of ATP13A2 on α-synuclein multimerization via polyamine transport dependent and independent functions.

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