Inhibition of Vibrio anguillarum byPseudomonas fluorescens AH2, a Possible Probiotic Treatment of Fish

ABSTRACT To study the possible use of probiotics in fish farming, we evaluated the in vitro and in vivo antagonism of antibacterial strainPseudomonas fluorescens strain AH2 against the fish-pathogenic bacterium Vibrio anguillarum. As iron is important in virulence and bacterial interactions, the effect ofP. fluorescens AH2 was studied under iron-rich and iron-limited conditions. Sterile-filtered culture supernatants from iron-limited P. fluorescens AH2 inhibited the growth ofV. anguillarum, whereas sterile-filtered supernatants from iron-replete cultures of P. fluorescens AH2 did not.P. fluorescens AH2 inhibited the growth of V. anguillarum during coculture, independently of the iron concentration, when the initial count of the antagonist was 100 to 1,000 times greater that of the fish pathogen. These in vitro results were successfully repeated in vivo. A probiotic effect in vivo was tested by exposing rainbow trout (Oncorynchus mykissWalbaum) to P. fluorescens AH2 at a density of 105 CFU/ml for 5 days before a challenge with V. anguillarum at 104 to 105 CFU/ml for 1 h. Some fish were also exposed to P. fluorescens AH2 at 107 CFU/ml during the 1-h infection. The combined probiotic treatment resulted in a 46% reduction of calculated accumulated mortality; accumulated mortality was 25% after 7 days at 12°C in the probiotic-treated fish, whereas mortality was 47% in fish not treated with the probiont.

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Molecular characterization of the TonB2 protein from the fish pathogen Vibrio anguillarum

Biochemical Journal ◽

10.1042/bj20081462 ◽

2009 ◽

Vol 418 (1) ◽

pp. 49-59 ◽

Cited By ~ 14

Author(s):

Claudia S. López ◽

R. Sean Peacock ◽

Jorge H. Crosa ◽

Hans J. Vogel

Keyword(s):

Amino Acids ◽

Solution Structure ◽

Bacterial Species ◽

Vibrio Anguillarum ◽

Fish Pathogen ◽

E Coli ◽

Terminal Domain ◽

C Terminus

In the fish pathogen Vibrio anguillarum the TonB2 protein is essential for the uptake of the indigenous siderophore anguibactin. Here we describe deletion mutants and alanine replacements affecting the final six amino acids of TonB2. Deletions of more than two amino acids of the TonB2 C-terminus abolished ferric-anguibactin transport, whereas replacement of the last three residues resulted in a protein with wild-type transport properties. We have solved the high-resolution solution structure of the TonB2 C-terminal domain by NMR spectroscopy. The core of this domain (residues 121–206) has an αββαβ structure, whereas residues 76–120 are flexible and extended. This overall folding topology is similar to the Escherichia coli TonB C-terminal domain, albeit with two differences: the β4 strand found at the C-terminus of TonB is absent in TonB2, and loop 3 is extended by 9 Å (0.9 nm) in TonB2. By examining several mutants, we determined that a complete loop 3 is not essential for TonB2 activity. Our results indicate that the β4 strand of E. coli TonB is not required for activity of the TonB system across Gram-negative bacterial species. We have also determined, through NMR chemical-shift-perturbation experiments, that the E. coli TonB binds in vitro to the TonB box from the TonB2-dependent outer membrane transporter FatA; moreover, it can substitute in vivo for TonB2 during ferric-anguibactin transport in V. anguillarum. Unexpectedly, TonB2 did not bind in vitro to the FatA TonB-box region, suggesting that additional factors may be required to promote this interaction. Overall our results indicate that TonB2 is a representative of a different class of TonB proteins.

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Effects of Enterococcus faecalis UGRA10 and the enterocin AS-48 against the fish pathogen Lactococcus garvieae. Studies in vitro and in vivo

Food Microbiology ◽

10.1016/j.fm.2018.08.002 ◽

2019 ◽

Vol 77 ◽

pp. 69-77 ◽

Cited By ~ 8

Author(s):

Alberto Baños ◽

Juan José Ariza ◽

Cristina Nuñez ◽

Lidia Gil-Martínez ◽

J. David García-López ◽

...

Keyword(s):

Enterococcus Faecalis ◽

Fish Pathogen ◽

Lactococcus Garvieae

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Evidence ofIn VivoProphage Induction during Clostridium difficile Infection

Applied and Environmental Microbiology ◽

10.1128/aem.02275-12 ◽

2012 ◽

Vol 78 (21) ◽

pp. 7662-7670 ◽

Cited By ~ 58

Author(s):

Mathieu Meessen-Pinard ◽

Ognjen Sekulovic ◽

Louis-Charles Fortier

Keyword(s):

Clostridium Difficile ◽

Bioinformatics Analyses ◽

Content Type ◽

Unique Character ◽

Viral Particles ◽

Potential Impact ◽

First Time ◽

Culture Supernatants

ABSTRACTProphages contribute to the evolution and virulence of most bacterial pathogens, but their role inClostridium difficileis unclear. Here we describe the isolation of fourMyoviridaephages, ϕMMP01, ϕMMP02, ϕMMP03, and ϕMMP04, that were recovered as free viral particles in the filter-sterilized stool supernatants of patients suffering fromC. difficileinfection (CDI). Furthermore, identical prophages were found in the chromosomes ofC. difficileisolated from the corresponding fecal samples. We therefore provide, for the first time, evidence ofin vivoprophage induction during CDI. We completely sequenced the genomes of ϕMMP02 and ϕMMP04, and bioinformatics analyses did not reveal the presence of virulence factors but underlined the unique character of ϕMMP04. We also studied the mobility of ϕMMP02 and ϕMMP04 prophagesin vitro. Both prophages were spontaneously induced, with 4 to 5 log PFU/ml detected in the culture supernatants of the corresponding lysogens. When lysogens were grown in the presence of subinhibitory concentrations of ciprofloxacin, moxifloxacin, levofloxacin, or mitomycin C, the phage titers further increased, reaching 8 to 9 log PFU/ml in the case of ϕMMP04. In summary, our study highlights the extensive genetic diversity and mobility ofC. difficileprophages. Moreover, antibiotics known to represent risk factors for CDI, such as quinolones, can stimulate prophage mobilityin vitroand probablyin vivoas well, which underscores their potential impact on phage-mediated horizontal gene transfer events and the evolution ofC. difficile.

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Ferritin accumulation and degradation in different organs of pea (Pisum sativum) during development

Biochemical Journal ◽

10.1042/bj2740601 ◽

1991 ◽

Vol 274 (2) ◽

pp. 601-606 ◽

Cited By ~ 98

Author(s):

S Lobreaux ◽

J F Briat

Keyword(s):

Pisum Sativum ◽

Total Protein ◽

Iron Concentration ◽

Embryo Axis ◽

Iron Supply ◽

Hydroponic Cultures ◽

Radical Damage ◽

Pea Seed

Iron concentration and ferritin distribution have been determined in different organs of pea (Pisum sativum) during development under conditions of continuous iron supply from hydroponic cultures. No ferritin was detected in total protein extracts from roots or leaves. However, a transient iron accumulation in the roots, which corresponds to an increase in iron uptake, was observed when young fruits started to develop. Ferritin was detectable in total protein extracts of flowers and pods, and it accumulated in seeds. In seeds, the same relative amount of ferritin was detected in cotyledons and in the embryo axis. In cotyledons, ferritin and iron concentration decrease progressively during the first week of germination. Ferritin in the embryo axis was processed, and disappeared, during germination, within the first 4 days of radicle and epicotyl growth. This degradation of ferritin in vivo was marked by a shortening of a 28 kDa subunit, giving 26.5 and 25 kDa polypeptides, reminiscent of the radical damage occurring in pea seed ferritin during iron exchange in vitro [Laulhere, Laboure & Briat (1989) J. Biol. Chem. 264, 3629-3635]. Developmental control of iron concentration and ferritin distribution in different organs of pea is discussed.

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Antimicrobial Peptides Protect Coho Salmon fromVibrio anguillarum Infections

Applied and Environmental Microbiology ◽

10.1128/aem.66.5.1928-1932.2000 ◽

2000 ◽

Vol 66 (5) ◽

pp. 1928-1932 ◽

Cited By ~ 80

Author(s):

X. Jia ◽

A. Patrzykat ◽

R. H. Devlin ◽

P. A. Ackerman ◽

G. K. Iwama ◽

...

Keyword(s):

Antimicrobial Peptides ◽

Coho Salmon ◽

Vibrio Anguillarum ◽

Transgenic Fish ◽

In Vivo Studies ◽

Bacterial Disease ◽

Control Groups ◽

Fish Pathogens

ABSTRACT Fish losses from infectious diseases are a significant problem in aquaculture worldwide. Therefore, we investigated the ability of cationic antimicrobial peptides to protect against infection caused by the fish pathogen Vibrio anguillarum. To identify effective peptides for fish, the MICs of certain antimicrobial peptides against fish pathogens were determined in vitro. Two of the most effective antimicrobial peptides, CEME, a cecropin-melittin hybrid peptide, and pleurocidin amide, a C-terminally amidated form of the natural flounder peptide, were selected for in vivo studies. A single intraperitoneal injection of CEME did not affect mortality rates in juvenile coho salmon infected with V. anguillarum, the causative agent of vibriosis. Therefore, the peptides were delivered continuously using miniosmotic pumps placed in the peritoneal cavity. Twelve days after pump implantation, the fish received intraperitoneal injections ofV. anguillarum at a dose that would kill 50 to 90% of the population. Fish receiving 200 μg of CEME per day survived longer and had significantly lower accumulated mortalities (13%) than the control groups (50 to 58%). Fish receiving pleurocidin amide at 250 μg per day also survived longer and had significantly lower accumulated mortalities (5%) than the control groups (67 to 75%). This clearly shows the potential for antimicrobial peptides to protect fish against infections and indicates that the strategy of overexpressing the peptides in transgenic fish may provide a method of decreasing bacterial disease problems.

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Immune response-associated production of neopterin. Release from macrophages primarily under control of interferon-gamma.

Journal of Experimental Medicine ◽

10.1084/jem.160.1.310 ◽

1984 ◽

Vol 160 (1) ◽

pp. 310-316 ◽

Cited By ~ 718

Author(s):

C Huber ◽

J R Batchelor ◽

D Fuchs ◽

A Hausen ◽

A Lang ◽

...

Keyword(s):

Immune Response ◽

Immune Responses ◽

Active Principle ◽

Activated T Cells ◽

Cellular Origin ◽

Associated Production ◽

Neurotransmitter Synthesis ◽

Culture Supernatants

Neopterin, a compound derived from GTP, represents a precursor molecule of biopterin that is an essential cofactor in neurotransmitter synthesis. We have recently reported that in vivo as well as in vitro immune responses are accompanied by an increased release of neopterin and that this phenomenon can be used for the biochemical monitoring of diseases accompanied by hyperimmune stimulation. This article deals with the cellular origin and the control of this immune response-associated neopterin release in vitro. Using highly purified or monoclonal cellular reagents we demonstrate that macrophages (M phi) stimulated with supernatants from activated T cells release large amounts of neopterin into culture supernatants. Further experiments involving induction of neopterin release from M phi with various human recombinant interferons (IFNs) or neutralization of the effect of T cell supernatants with various monoclonal anti-IFN antibodies revealed immune IFN as the active principle. It thus appears that a metabolic pathway so far exclusively known in context with the generation of an essential cofactor of neurotransmitter-synthesis during immune responses is also activated in M phi under stringent control by immune IFN-like lymphokines.

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beta-Naphthoflavone abolishes interrenal sensitivity to ACTH stimulation in rainbow trout

Journal of Endocrinology ◽

10.1677/joe.0.1570063 ◽

1998 ◽

Vol 157 (1) ◽

pp. 63-70 ◽

Cited By ~ 40

Author(s):

JM Wilson ◽

MM Vijayan ◽

CJ Kennedy ◽

GK Iwama ◽

TW Moon

Keyword(s):

Cytochrome P450 ◽

Rainbow Trout ◽

Acth Stimulation ◽

Rainbow Trout Oncorhynchus Mykiss ◽

Treated Fish ◽

First Time ◽

Hepatic Cytochrome ◽

Stimulation Of

We report for the first time that beta-naphthoflavone (BNF) abolishes ACTH stimulation of cortisol production in rainbow trout (Oncorhynchus mykiss). There was significantly higher hepatic cytochrome P450 content and ethoxyresorufin O-de-ethylase and uridine-5'-diphosphoglucuronic acid transferase activities in BNF-treated fish than in sham-treated controls. BNF did not significantly affect either plasma turnover or tissue distribution of [3H]cortisol-derived radioactivity. Hepatic membrane fluidity and hepatocyte capacity for cortisol uptake were not altered by BNF as compared with the sham-treated fish. These results taken together suggest that BNF does not affect cortisol-clearance mechanisms in trout. A 3 min handling disturbance period elicited a plasma cortisol response in the sham-treated fish; however, the response in the BNF-treated fish was muted and significantly lower than in the sham fish. This in vivo response corroborates the lack of interrenal sensitivity to ACTH in vitro in the BNF-treated fish, suggesting that BNF affects the ACTH pathway in trout. Our results suggest the possibility that cytochrome P450-inducing compounds may affect cortisol dynamics by decreasing interrenal responsiveness to ACTH stimulation in fish, thereby impairing the physiological responses that are necessary for the animal to cope with the stressor.

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Integrated Optimization of the In Vivo Heme Biosynthesis Pathway and the In Vitro Iron Concentration for 5-Aminolevulinate Production

Applied Biochemistry and Biotechnology ◽

10.1007/s12010-015-1942-2 ◽

2015 ◽

Vol 178 (6) ◽

pp. 1252-1262 ◽

Cited By ~ 6

Author(s):

Junli Zhang ◽

Zhen Kang ◽

Wenwen Ding ◽

Jian Chen ◽

Guocheng Du

Keyword(s):

Iron Concentration ◽

Heme Biosynthesis ◽

Biosynthesis Pathway ◽

Integrated Optimization

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Combination of garlic - shatterstone herb powder to control Streptococcus agalactiae infection in tilapia

Jurnal Akuakultur Indonesia ◽

10.19027/jai.14.79-89 ◽

2015 ◽

Vol 14 (1) ◽

pp. 79

Author(s):

Ririn Nurul Fauziah ◽

Dinamella Wahjuningrum ◽

, Sukenda ◽

, Ranta

Keyword(s):

Growth Rate ◽

Streptococcus Agalactiae ◽

Allium Sativum ◽

Positive Control ◽

Quality Parameters ◽

Negative Control ◽

Fish Pathogen ◽

Phyllanthus Niruri

ABSTRACT This study was aimed at determining potential of combination powder of garlic Allium sativum-shatterstone herb Phyllanthus niruri supplemented in feed against S. agalactiae infection in tilapia. Four concentrations of combination powder of A. sativum-P. Niruri; 20+5, 20+10, 20+15 and 20+20 ppt respectively were investigated for their ability to inhibit bacterial fish pathogen. Combination dose of 20+15 ppt produced the highest inhibitory zones in in vitro test. In vivo test consisted of three treatments with three replications, namely positive control (K+), negative control (K-) and the treatment of A. sativum-P. niruri suplemented in feed (BM). The test perfomed on tilapia with weight of 10.33 ± 1.63 g and were reared at density of 10 ind/aquarium. The fish was fed for 14 days, then injected intraperitoneally with 0.1 mL S. agalactiae at concentration of 105 cfu/mL for positive control and BM groups. Survival, growth rate, feed response, hematological and water quality parameters were observed for 10 days. This study showed that the suplemented-feed-fish (BM) showed better growth rate, feed response, and survival (83.3%) than positive control (36.7%) at P<0.05. In addition, A. sativum-P. niruri suplemented in feed was also able to enhance the immune response by increasing phagocytic activity. Keywords: Streptococcus agalactiae, phytopharmacy, Allium sativum-Phyllanthus niruri, tilapia ABSTRAK Penelitian ini bertujuan untuk menganalisis potensi campuran tepung bawang putih Allium sativum-meniran Phyllanthus niruri dalam pakan terhadap pencegahan infeksi bakteri S. agalactiae pada ikan nila. Empat konsentrasi campuran tepung bawang putih-meniran yaitu 20+5 ppt, 20+10 ppt, 20+15 ppt dan 20+20 ppt masing-masing diuji kemampuannya dalam menghambat bakteri patogen pada ikan. Campuran dosis 20+15 ppt menghasilkan zona hambat terbaik dalam uji in vitro. Uji in vivo terdiri atas tiga perlakuan dengan tiga ulangan yaitu kontrol positif, kontrol negatif, dan perlakuan pakan yang mengandung bawang putih-meniran (BM). Uji ini dilakukan pada ikan nila berbobot 10,33±1,63 g yang dipelihara di akuarium dengan kepadatan 10 ekor/akuarium. Ikan diberi pakan perlakuan selama 14 hari kemudian diinjeksi secara intraperitoneal dengan bakteri S. agalactiae sebanyak 0,1 mL dengan kepadatan 105 cfu/mL pada perlakuan kontrol positif dan perlakuan BM. Parameter kelangsungan hidup, laju pertumbuhan, respons pakan, parameter hematologi, dan kualitas air diamati selama sepuluh hari. Hasil dari penelitian ini menunjukkan bahwa pemberian BM dalam pakan memberikan laju pertumbuhan, respons pakan, dan sintasan (83,3%) yang lebih baik daripada kontrol positif (36,7%) pada P<0,05. Pakan yang mengandung campuran bawang putih-meniran ini juga mampu meningkatkan respons imun dengan adanya peningkatan aktivitas fagositosis. Kata kunci: Streptococcus agalactiae, fitofarmaka, Allium sativum-Phyllanthus niruri, ikan nila

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Screening of Probiotic Bacteria from Intestine and Culture Environment of Hoeven’s slender carp Leptobarbus hoeveni Blkr to Control Pathogenic Bacteria

Jurnal Akuakultur Indonesia ◽

10.19027/jai.9.127-135 ◽

2010 ◽

Vol 9 (2) ◽

pp. 127

Author(s):

. Sunarto ◽

. Sukenda ◽

. Widanarni

Keyword(s):

Aeromonas Hydrophila ◽

Pathogenic Bacteria ◽

Probiotic Bacteria ◽

Streptococcus Iniae ◽

Mycobacterium Fortuitum ◽

Fish Pathogen ◽

Culture Environment ◽

Control Disease

The ability of probiotic bacteria to control disease infection has been used in aquaculture. This experiment was conducted to isolate and characterize probiotic bacteria; the competition test its ability probiotic bacteria against pathogenic bacteria; and to improve survival rate of Leptobarbus hoeveni. The bacteria were isolated from Leptobarbus hoeveni and its culture environment, and then tested to know its ability to inhibit bacterial fish pathogen in-vitro. Furthermore, the selected probiotic bacteria were tested in vivo to evaluate their ability to inhibit pathogen of Leptobarbus hoeveni. The result showed that probiotic bacteria inhibit the growth of Streptococcus iniae, Flexibacter columnaris, Mycobacterium fortuitum and Aeromonas hydrophila in vitro. Isolate DD3 was the best of candidate probiotic because of the ability to inhibit pathogen, especially A. hydrophila, the most virulent bacteria in Leptobarbus hoeveni. Key Words : probiotic bacteria, Leptobarbus hoeveni, pathogenic bacteria Abstrak Kemampuan bakteri probiotik untuk mengendalikan penyakit infeksi telah digunakan dalam akuakultur. Tujuan penelitian ini adalah mengisolasi dan mengkarakterisasi bakteri probiotik, menguji kemampuan bakteri probiotik terhadap bakteri patogen, sehingga dapat meningkatkan tingkat kelangsungan hidup ikan jelawat. Bakteri diisolasi dari usus ikan jelawat dan lingkungan budaya, kemudian diuji kemampuannya menghambat bakteri patogen secara in-vitro. Selanjutnya bakteri probiotik yang dipilih diuji secara in vivo untuk mengevaluasi kemampuannya dalam menghambat patogen di dalam tubuh ikan jelawat. Dari hasil penelitian diperoleh bakteri probiotik yang diisolasi dari usus dan lingkungan budaya ikan jelawat menunjukkan penghambatan pertumbuhan terhadap Streptococcus iniae, Flexibacter columnaris, Mycobacterium fortuitum dan Aeromonas hydrophila secara in vitro. Isolat DD3 merupakan kandidat probiotik terbaik, karena mempunyai kemampuan untuk menghambat bakteri patogen, khususnya bakteri A. hydrophila adalah bakteri yang paling viluren bagi ikan jelawat. Kata Kunci: bakteri probiotik, ikan jalawat dan baktri patogen

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