Coculture of Acinetobacter johnsonii and Shewanella putrefaciens Contributes to the ABC Transporter that Impacts Cold Adaption in the Aquatic Food Storage Environment.

Acinetobacter johnsonii and Shewanella putrefaciens were identified as specific spoilage organisms in aquatic food. The interactions among specific spoilage organisms under cold stress have a significant impact on the assembly of microbial communities, which play crucial roles in the spoilage and cold adaptation processes. The limited understanding of A. johnsonii and S. putrefaciens interactions in the cold adaptation mechanism hinders the elucidation of their roles in protein and metabolism levels. 4D quantitative proteomic analysis showed that the coculture of A. johnsonii and S. putrefaciens responds to low temperatures through ABC transporter proteins, resulting in phospholipid transport and inner membrane components. SapA and FtsX proteins were significantly upregulated, while LolC, LolD, LolE, PotD, PotA, PotB, and PotC proteins were significantly downregulated. Metabolome assays revealed that metabolites of glutathione and spermidine/putrescin were significantly upregulated, while metabolites of arginine/lysine/ornithine were significantly downregulated and involved in the ABC transporter metabolism. The results of ultramicroscopic analyses showed that the coculture of A. johnsonii and S. putrefaciens surface combined with the presence of the leakage of intracellular contents, suggesting that the bacteria were severely damaged and wrinkled to absorb metabolic nutrients and adapt to cold temperatures.

Influence of marine Shewanella putrefaciens and mediated calcium deposition on Q235 carbon steel corrosion.

The microbiologically influenced corrosion inhibition (MICI) of Q235 carbon steel by Shewanella putrefaciens and mediated calcium deposition were investigated by regulating microbial mineralization. In a calcium-rich medium, S. putrefaciens rapidly created a protective calcium carbonate layer on the steel surface, which blocked Cl- diffusion. Without calcium, the biofilm and rust layer mitigated pitting corrosion but did not prevent Cl- penetration. Potentiodynamic polarization results indicated that the current densities (icorr values) of the corrosion produced in the S. putrefaciens-inoculated media with and without calcium were 0.4 µA/cm2 and 0.6 µA/cm2, respectively. Similarly, compared with those under sterile conditions, the corrosion inhibition rates were 92.2% and 87.4% higher, respectively. Electrochemical impedance spectroscopy (EIS) and scanning electrochemical microscopy (SECM) confirmed that the MICI was caused by the combination of microbial aerobic respiration and the deposited layers. Even under nonbiological conditions, S. putrefaciens-induced calcium carbonate deposition inhibited corrosion.

Evaluation of the Differential Postbiotic Potential of Shewanella putrefaciens Pdp11 Cultured in Several Growing Conditions.

The increased knowledge of functional foods has led to the development of a new generation of health products, including those containing probiotics and products derived from them. Shewanella putrefaciens Pdp11 (SpPdp11) is a strain described as a probiotic that exerts important beneficial effects on several farmed fish. However, the use of live probiotic cells in aquaculture has limitations such as uncertain survival and shelf life, which can limit their efficacy. In addition, its efficacy can vary across species and hosts. When probiotics are administered orally, their activity can be affected by the environment present in the host and by interactions with the intestinal microbiota. Furthermore, live cells can also produce undesired substances that may negatively impact the host as well as the risk of potential virulence reversion acquired such as antibiotic resistance. Therefore, new alternatives emerged such as postbiotics. Currently, there is no knowledge about the postbiotic potential of SpPdp11 in the aquaculture industry. Postbiotic refers to the use of bacterial metabolites, including extracellular products (ECPs), to improve host physiology. However, the production of postbiotic metabolites can be affected by various factors such as cultivation conditions, which can affect bacterial metabolism. Thus, the objective of this study was to evaluate the postbiotic potential of ECPs from SpPdp11 under different cultivation conditions, including culture media, temperature, growth phase, and salinity. We analyzed their hydrolytic, antibacterial, antiviral, and cytotoxic capacity on several fish cell lines. The results obtained have demonstrated how each ECP condition can exert a different hydrolytic profile, reduce the biofilm formation by bacterial pathogens relevant to fish, lower the titer of nervous necrosis virus (NNV), and exert a cytotoxic effect on different fish cell lines. In conclusion, the ECPs obtained from SpPdp11 have different capacities depending on the cultivation conditions used. These conditions must be considered in order to recover the maximum number of beneficial capacities or to choose the appropriate conditions for specific activities.

Optimisation of treatment conditions for reducing Shewanella putrefaciens and Salmonella Typhimurium on grass carp treated by thermoultrasound-assisted plasma functionalized buffer.

A novel method of thermoultrasound-assisted plasma functionalized buffer (PFB) for decontaminating grass carp was evaluated using the Box-Behnken design (BBD) with processing variables including PFB generating voltage (PV), ultrasound treatment time (UT) and temperature (TP). The predicted models were found to be significant (p < 0.05) and displayed sufficient fitness with experimental data as indicated by non-significant (p > 0.05) lack of fit and high coefficient of determination (R2≥0.97) values. The optimum decontamination conditions for the responses of S. putrefaciens and S. Typhimurium were PV of 66 V, UT of 14.90 min and TP of 60 ℃, achieving reductions of 4.40 and 3.97 log CFU/g, respectively, with a desirability of 0.998. Among the variables, temperature presented higher significance for inactivating bacteria and the production of volatile basic nitrogen and lipid peroxidation under the optimized conditions were within the limits of freshness for grass carp. Additionally, the effects of PFB and the optimized thermoultrasound-assisted PFB decontamination were mild on the microstructure of grass carp with slight ruptures and loose myofibril structures, indicating the potential of thermoultrasound-assisted PFB for seafood products decontamination with reduced processing time.

Formation and function of bacterial organelles.

Advances in imaging technologies have revealed that many bacteria possess organelles with a proteomically defined lumen and a macromolecular boundary. Some are bound by a lipid bilayer (such as thylakoids, magnetosomes and anammoxosomes), whereas others are defined by a lipid monolayer (such as lipid bodies), a proteinaceous coat (such as carboxysomes) or have a phase-defined boundary (such as nucleolus-like compartments). These diverse organelles have various metabolic and physiological functions, facilitating adaptation to different environments and driving the evolution of cellular complexity. This Review highlights that, despite the diversity of reported organelles, some unifying concepts underlie their formation, structure and function. Bacteria have fundamental mechanisms of organelle formation, through which conserved processes can form distinct organelles in different species depending on the proteins recruited to the luminal space and the boundary of the organelle. These complex subcellular compartments provide evolutionary advantages as well as enabling metabolic specialization, biogeochemical processes and biotechnological advances. Growing evidence suggests that the presence of organelles is the rule, rather than the exception, in bacterial cells.

Dietary administration of the probiotic Shewanella putrefaciens to experimentally wounded gilthead seabream (Sparus aurata L.) facilitates the skin wound healing.

The effect of the probiotic Shewanella putrefaciens Pdp11 (SpPdp11) was studied on the skin healing of experimentally wounded gilthead seabream (Sparus aurata L.). Two replicates (n = 12) of fish were fed CON diet or SP diet for 30 days. Half of the fish were sampled while the others were injured and sampled 7 days post-wounding. Results by image analysis of wound areas showed that SpPdp11 inclusion facilitated wound closure. Compared with the CON group, fish in SP group sampled 7 days post-wounding had a significantly decreased serum AST and increased ALB/GLOB ratio. Furthermore, protease and peroxidase activities were significantly increased in skin mucus from fish in SP group sampled 7 days post-wounding, compared with those fed CON diet. Additionally, SP diet up-regulated the gene expression of antioxidant enzymes, anti-inflammatory cytokines, and re-epithelialization related genes in the fish skin. Furthermore, significant decreases in pro-inflammatory cytokines expression were detected in fish from SP group, respect to control ones. Overall, SpPdp11 inclusion facilitated the wound healing and the re-epithelialization of the damaged skin, alleviated the inflammatory response in the wound area through intensifying the antioxidant system, and enhancing the neo-vascularization and the synthesis of matrix proteins in the skin wound sites of fish.

Electrical tension-triggered conversion of anaerobic to aerobic respiration of Shewanella putrefaciens CN32 cells while promoting biofilm growth in microbial fuel cells.

A global gene expression analysis of Shewanella putrefaciens CN32 cells nearby a nanostructured microbial anode reveals an electrical tension-triggered conversion of anaerobic respiration to aerobic respiration with increased excretion of flavin electron shuttles and cytochrome C proteins, which sheds light on the role of electric tension in cell organisms.

Influence of skin wounds on the intestinal inflammatory response and barrier function: Protective role of dietary Shewanella putrefaciens SpPdp11 administration to gilthead seabream (Sparus aurata L.).

The effects of skin wounds on the intestinal barrier function and the beneficial effects of the dietary administration of Shewanella putrefaciens (known as SpPdp11) in gilthead seabream (Sparus aurata L.) were studied. Two replicates of fish were fed a commercial diet (control, CON) or CON diet enriched with 109 cfu g-1 SpPdp11 (SP diet) for 30 days. After this time, half of the fish were sampled, while the others were injured below the lateral line (wounded fish, W) and fed the same diets for an extra week before sampling (CON + W and SP + W groups). The intestinal histology and gene expression of different genes relevant for the intestinal barrier function were studied. The results showed that injured fish had a disordered enterocyte nucleus disposition, a more intense infiltration of mixed leucocytes and a thicker lamina propria in the intestine compared to the control fish. However, the fish in the SP + W group did not present these pathological symptoms in the intestine. No significant variations in the number of goblet cells were detected among the different experimental groups. Pro-inflammatory cytokines (colony-stimulating factor receptor 1, CSF1R, myeloperoxidase, MPO and interleukin-1ß, IL-1ß), mucins (intestinal mucin, IMUC and mucin 2, MUC2), and immunoglobulin T heavy chain (IGHT) were up-regulated, while tight junction protein occludin was down-regulated in the intestine from fish of the CON + W group. Similarly, the dietary administration of SpPdp11 markedly depressed the gene expression of pro-inflammatory cytokines, MUC2 and IGHT, but increased the gene expression of anti-inflammatory cytokine transforming growth factor-ß1 (TGF-ß1) and the tight junction proteins tricellulin and occluding after wounding. In brief, the skin wounds provoked an intestinal inflammatory response that included changes in the mucus layer and tight junction disruptions. Besides this, preventive administration of SpPdp11 alleviated the intestinal dysfunctions caused by skin wounds in gilthead seabream.

Enhanced extracellular electron transfer between Shewanella putrefaciens and carbon felt electrode modified by bio-reduced graphene oxide.

Extracellular electron transfer (EET) is a governing factor for the electrochemical performance of a bioelectrochemical system (BES) such as the microbial fuel cell (MFC). Herein, an in situ method to fabricate a bio-reduced graphene oxide (GO) (br-GO) modified carbon felt electrode to increase EET was developed. GO (0.5mgmL-1) was spiked into the anode chamber in a three-electrode BES and was transformed to br-GO with a self-assembled three-dimensional (3D) structure. The response of the br-GO modified electrode potential to the attached population of Shewanella putrefaciens increased from 0.071V to 0.517V (vs Ag/AgCl). Meanwhile, br-GO modification resulted a significant enhancement in the total amount of extracellular electrons transferred between the modified electrode and microbe. The process of br-GO modification lowered the charge transfer resistance of the electrode and enhanced the EET. The modified electrode was further employed as an anode in the MFC, and consequently, the power density of the MFC was significantly enhanced. The current study not only gives a simple and effective way for improving the EET with br-GO fabrication, but also provides a strategy to enhance the power density of the MFC.

Characterization and antimicrobial mechanism of CF-14, a new antimicrobial peptide from the epidermal mucus of catfish.

In this study, we identified a novel antibacterial peptide, RIVELTLPRVSVRL-NH2 (named CF-14), derived from the epidermal mucus of catfish and characterized its antimicrobial activity. Analysis of antimicrobial activity and hemolytic activity of CF-14 revealed broad spectrum, high levels of antimicrobial activity and low toxicity to eukaryotic cells. CF-14 remained stable at pH values ranging from 4.0 to 12.0 and remained bioactive when exposed to high temperature. CD analysis indicated that CF-14 forms a random coil in PBS buffer and an α-helical conformation in the membrane-mimetic 2.5% SDS micelle. Additionally, the antibacterial mechanism of CF-14 against Shewanella putrefaciens was investigated. Membrane permeability experiments confirmed that CF-14 could increase cell wall membrane permeability and cause nucleotide leakage. Moreover, observations performed using scanning electron and confocal microscopy indicated that CF-14 could penetrate into the cell membranes of S. putrefaciens and accumulate in bacterial cells, but did not break down cell membranes. Further, electrophoresis analysis demonstrated that CF-14 possesses DNA-binding affinity. The results provide a substantial basis for future application of CF-14, a novel cell-penetrating peptide (CPP) derived from catfish.

Synergistic action of Bacillus subtilis, Escherichia coli and Shewanella putrefaciens along with Pseudomonas putida on inhibiting mild steel against oxygen corrosion.

Microbial biofilm can effectively alter the electrochemical characteristics at metal/solution interface that can either accelerate or decelerate corrosion. The present paper reports about microbiologically induced corrosion inhibition (MICI) using Pseudomonas putida as a dominant bacterium under aerobic condition. Effective corrosion inhibition is achieved by the synergistic metabolic action of P. putida along with Escherichia coli, Bacillus subtilis or Shewanella putrefaciens. The synergistic metabolic actions of these bacteria in biopassivation are analysed with various aspects such as electrochemical impedance spectroscopy (EIS), Fourier transform infrared spectroscopy (FT-IR), field emission scanning electron microscopy (FESEM) and confocal laser scanning microscopy (CLSM). Surface topography is quantitatively analysed using optical scanning profilometry (OSP). The binary culture system containing P. putida + E. coli and P. putida + S. putrefaciens achieves an inhibition efficiency of 90% and 85% respectively, despite S. putrefaciens being a corrosion causing bacteria. The P. putida + E. coli system could form a stable biofilm on mild steel surface, with a high corrosion potential (- 329 mV vs. Ag/AgCl/KCl sat'd) and a low corrosion rate (1.65 × 10-1 mmpy). The presence of B. subtilis in the culture promotes corrosion against normal predictions. In the present case, the metabolic activities of the bacterial system on the mild steel surface cause depletion of oxygen in the medium that leads to suppression of corrosion. In addition, the biofilm could form an effective protective barrier on the metal surface that can suppress diffusion of corrosion products resulting in enhanced corrosion inhibition efficiency.

Spatial arrangement of several flagellins within bacterial flagella improves motility in different environments.

Bacterial flagella are helical proteinaceous fibers, composed of the protein flagellin, that confer motility to many bacterial species. The genomes of about half of all flagellated species include more than one flagellin gene, for reasons mostly unknown. Here we show that two flagellins (FlaA and FlaB) are spatially arranged in the polar flagellum of Shewanella putrefaciens, with FlaA being more abundant close to the motor and FlaB in the remainder of the flagellar filament. Observations of swimming trajectories and numerical simulations demonstrate that this segmentation improves motility in a range of environmental conditions, compared to mutants with single-flagellin filaments. In particular, it facilitates screw-like motility, which enhances cellular spreading through obstructed environments. Similar mechanisms may apply to other bacterial species and may explain the maintenance of multiple flagellins to form the flagellar filament.

ZomB is essential for flagellar motor reversals in Shewanella putrefaciens and Vibrio parahaemolyticus.

The ability of most bacterial flagellar motors to reverse the direction of rotation is crucial for efficient chemotaxis. In Escherichia coli, motor reversals are mediated by binding of phosphorylated chemotaxis protein CheY to components of the flagellar rotor, FliM and FliN, which induces a conformational switch of the flagellar C-ring. Here, we show that for Shewanella putrefaciens, Vibrio parahaemolyticus and likely a number of other species an additional transmembrane protein, ZomB, is critically required for motor reversals as mutants lacking ZomB exclusively exhibit straightforward swimming also upon full phosphorylation or overproduction of CheY. ZomB is recruited to the cell poles by and is destabilized in the absence of the polar landmark protein HubP. ZomB also co-localizes to and may thus interact with the flagellar motor. The ΔzomB phenotype was suppressed by mutations in the very C-terminal region of FliM. We propose that the flagellar motors of Shewanella, Vibrio and numerous other species harboring orthologs to ZomB are locked in counterclockwise rotation and may require interaction with ZomB to enable the conformational switch required for motor reversals. Regulation of ZomB activity or abundance may provide these species with an additional means to modulate chemotaxis efficiency.

Proteomic assessment of the role of N-acyl homoserine lactone in Shewanella putrefaciens spoilage.

Shewanella spp. are the common spoilage organisms found in aquatic food products stored at low temperature and their spoilage mechanism has been reported to be mediated by quorum sensing (QS). However, the specifically expressed proteins responding to N-acyl homoserine-lactone (AHLs) were seldom reported. This study aims to evaluate the effects of different AHL signal molecules on Shewanella putrefaciens Z4 isolated from refrigerated turbot (Scophthalmus maximus) at the proteome level. The results revealed that exogenous AHLs were utilized as QS signal molecules by S. putrefaciens Z4, and AHLs were not degraded by intracellular or extracellular enzymes secreted by S. putrefaciens Z4. Twenty-three differently expressed spots upon the addition of AHLs were selected and identified by liquid chromatography-mass spectrometry (LC-MS). The results indicated that proteins involving in growth and metabolism (i.e. citrate synthase, succinate semialdehyde dehydrogenase), environment adaptation and regulators (i.e. polysaccharide deacetylases, transaldolase) were down-regulated upon three kinds of AHLs (C4-HSL, C6-HSL and O-C6-HSL), whereas the abundance of stress response protein and DNA ligase were elevated by the addition of exogenous AHLs. Moreover, the effects of exogenous C6-HSL and O-C6-HSLwere prominent. These results provide evidence that AHL-based QS signal molecules affected some important metabolic properties of S. putrefaciens. SIGNIFICANCE AND IMPACT OF THE STUDY: N-acyl homoserine lactone (AHLs)-based quorum sensing signal molecules involving in the behavior regulation in most of the Gram-negative bacteria have widely been reported. This study aims to evaluate the effect of AHLs on Shewanella putrefaciens Z4 at the proteome level. It provides the theoretic basis for elucidating the spoilage mechanism of Shewanella spp., the common spoilage micro-organism in refrigerated marine aquatic food products.

Bacteria exploit a polymorphic instability of the flagellar filament to escape from traps.

Many bacterial species swim by rotating single polar helical flagella. Depending on the direction of rotation, they can swim forward or backward and change directions to move along chemical gradients but also to navigate their obstructed natural environment in soils, sediments, or mucus. When they get stuck, they naturally try to back out, but they can also resort to a radically different flagellar mode, which we discovered here. Using high-speed microscopy, we monitored the swimming behavior of the monopolarly flagellated species Shewanella putrefaciens with fluorescently labeled flagellar filaments at an agarose-glass interface. We show that, when a cell gets stuck, the polar flagellar filament executes a polymorphic change into a spiral-like form that wraps around the cell body in a spiral-like fashion and enables the cell to escape by a screw-like backward motion. Microscopy and modeling suggest that this propagation mode is triggered by an instability of the flagellum under reversal of the rotation and the applied torque. The switch is reversible and bacteria that have escaped the trap can return to their normal swimming mode by another reversal of motor direction. The screw-type flagellar arrangement enables a unique mode of propagation and, given the large number of polarly flagellated bacteria, we expect it to be a common and widespread escape or motility mode in complex and structured environments.

Sodium Lactate Negatively Regulates Shewanella putrefaciens CN32 Biofilm Formation via a Three-Component Regulatory System (LrbS-LrbA-LrbR).

The capability of biofilm formation has a major impact on the industrial and biotechnological applications of Shewanella putrefaciens CN32. However, the detailed regulatory mechanisms underlying biofilm formation in this strain remain largely unknown. In the present report, we describe a three-component regulatory system which negatively regulates the biofilm formation of S. putrefaciens CN32. This system consists of a histidine kinase LrbS (Sputcn32_0303) and two cognate response regulators, including a transcription factor, LrbA (Sputcn32_0304), and a phosphodiesterase, LrbR (Sputcn32_0305). LrbS responds to the signal of the carbon source sodium lactate and subsequently activates LrbA. The activated LrbA then promotes the expression of lrbR, the gene for the other response regulator. The bis-(3'-5')-cyclic dimeric GMP (c-di-GMP) phosphodiesterase LrbR, containing an EAL domain, decreases the concentration of intracellular c-di-GMP, thereby negatively regulating biofilm formation. In summary, the carbon source sodium lactate acts as a signal molecule that regulates biofilm formation via a three-component regulatory system (LrbS-LrbA-LrbR) in S. putrefaciens CN32.IMPORTANCE Biofilm formation is a significant capability used by some bacteria to survive in adverse environments. Numerous environmental factors can affect biofilm formation through different signal transduction pathways. Carbon sources are critical nutrients for bacterial growth, and their concentrations and types significantly influence the biomass and structure of biofilms. However, knowledge about the underlying mechanism of biofilm formation regulation by carbon source is still limited. This work elucidates a modulation pattern of biofilm formation negatively regulated by sodium lactate as a carbon source via a three-component regulatory system in S. putrefaciens CN32, which may serve as a good example for studying how the carbon sources impact biofilm development in other bacteria.

Effect of zinc oxide film morphologies on the formation of Shewanella putrefaciens biofilm.

Zinc oxide (ZnO) films were prepared on aluminum substrate by a hydrothermal method to investigate the effect of their surface characteristics, including morphology and hydrophobicity, on the corresponding antibiofilm performance. The surface characteristics of the prepared ZnO films were examined by a comprehensive range of methodologies, suggesting that films of distinctive surface morphologies were successfully formed. Subsequently, their antibiofilm activities, using Shewanella putrefaciens as a model bacterium, were assessed. Surface measurements confirmed that the ZnO films equipped with a nanoscopic needlelike surface feature are more hydrophobic than those possessing densely packed microflakes. The reduced number of live cells and presence of biofilm, confirmed by optical and electron microscopy results, suggest that the former films possess an excellent antibiofilm performance. It is believed that the engineered nanoscopic needle feature might penetrate the cell membrane when they are in contact, allowing the effective substance of ZnO antibacterial ingredients to diffuse into the embedded bacteria. Furthermore, such surface characteristics might perturb the integrity of the cell membrane causing the intracellular substance is leaked from the cells. As such, the combinatorial effects of nanoscopic feature resulted in an inhibited growth of S. putrefaciens biofilm on ZnO film.

Boosting current generation in microbial fuel cells by an order of magnitude by coating an ionic liquid polymer on carbon anodes.

Microbial fuel cells (MFCs) have attracted great attentions due to their great application potentials, but the relatively low power densities of MFCs still hinder their widespread practical applications. Herein, we report that the current generation in MFCs can be boosted by an order of magnitude, simply by coating a hydrophilic and positively charged ionic liquid polymer (ILP) on carbon cloth (CC) or carbon felt (CF). The ILP coating not only can increase the bacterial loading capacity due to the electrostatic interactions between ILP and bacterial cells, but also can improve the mediated extracellular electron transfer between the electrode and the cytochrome proteins on the outer membrane of Shewanella putrefaciens cells. As a result, the maximum power density of a MFC equipped with the CF-ILP bioanode is as high as 4400±170mWm-2, which is amongst the highest values reported to date. This work demonstrates a new strategy for greatly boosting the current generation in MFCs.

An outbreak of Shewanella putrefaciens group in wild eels Anguilla anguilla L. favoured by hypoxic aquatic environments.

Microbiological analyses were conducted on wild eels from the L'Albufera Lake (Spain). A total of 174 individuals were collected in two surveys (i.e. year 2008 and autumn-winter 2014) among those caught by local fishermen into the lagoon. The prevalence of Shewanella putrefaciens group was 1.7% in 2008 and rose above 32% in 2014. It was due to an outbreak of shewanellosis that presented a morbidity rate of 64%. S. putrefaciens group strains were isolated as pure cultures from the sick eels that showed white ulcers surrounded by a reddish inflammation, damage of the mouth, extensive skin discoloration, exophthalmia, ascites and bad odour. The S. putrefaciens group was recovered from freshwater samples taken at the L'Albufera system, along autumn-winter 2015. Its counts significantly increased in freshwater parallel to hypoxia and temperature rising. Shewanellae strains were identified as S. putrefaciens and S. xiamenensis by 16S rRNA gene sequencing. These isolates recovered from sick eels or freshwater were virulent for European eel by IP challenge (LD50 106  CFU g-1 body weight). They also caused 30-38% cumulative mortality, in European eels challenged by a 2-h bath (107  CFU mL-1 ). These results suggest that shewanellosis could be transmitted through water highlighting the fact that hypoxic conditions increase this bacterium levels in water.

Antibiotic and heavy metal resistance in Shewanella putrefaciens strains isolated from shellfishes collected from West Sea, Korea.

The aim of the present study was to determine the antibiotic resistance patterns and distribution of heavy-metal resistance in Shewanella putrefaciens strains isolated from shellfishes collected from West Sea; and to determine the relationship, if any, between antibiotic and heavy-metal resistance in these strains. Among the 15 strains isolated, two strains, SY1 and SY2, showed heavy-metal resistance in addition to high resistance to seven antibiotics: cephalothin, gentamicin, erythromycin, vancomycin, ampicillin, rifampicin, and streptomycin. We conclude that heavy-metal contamination imposes long-term, widespread, and recalcitrant selection pressure, which potentially contributes to the maintenance and spread of antibiotic resistance factors in bacteria. Moreover, this fact holds both environmental and clinical importance.