Category Archives: Pseudomonas aeruginosa

Research – Opportunistic Pathogens in Drinking Water Distribution Systems—A Review

MDPI

Abstract

In contrast to “frank” pathogens, like Salmonella entrocoliticaShigella dysenteriae, and Vibrio cholerae, that always have a probability of disease, “opportunistic” pathogens are organisms that cause an infectious disease in a host with a weakened immune system and rarely in a healthy host. Historically, drinking water treatment has focused on control of frank pathogens, particularly those from human or animal sources (like Giardia lambliaCryptosporidium parvum, or Hepatitis A virus), but in recent years outbreaks from drinking water have increasingly been due to opportunistic pathogens. Characteristics of opportunistic pathogens that make them problematic for water treatment include: (1) they are normally present in aquatic environments, (2) they grow in biofilms that protect the bacteria from disinfectants, and (3) under appropriate conditions in drinking water systems (e.g., warm water, stagnation, low disinfectant levels, etc.), these bacteria can amplify to levels that can pose a public health risk. The three most common opportunistic pathogens in drinking water systems are Legionella pneumophilaMycobacterium avium, and Pseudomonas aeruginosa. This report focuses on these organisms to provide information on their public health risk, occurrence in drinking water systems, susceptibility to various disinfectants, and other operational practices (like flushing and cleaning of pipes and storage tanks). In addition, information is provided on a group of nine other opportunistic pathogens that are less commonly found in drinking water systems, including Aeromonas hydrophilaKlebsiella pneumoniaeSerratia marcescensBurkholderia pseudomalleiAcinetobacter baumanniiStenotrophomonas maltophiliaArcobacter butzleri, and several free-living amoebae including Naegleria fowleri and species of Acanthamoeba. The public health risk for these microbes in drinking water is still unclear, but in most cases, efforts to manage Legionella, mycobacteria, and Pseudomonas risks will also be effective for these other opportunistic pathogens. The approach to managing opportunistic pathogens in drinking water supplies focuses on controlling the growth of these organisms. Many of these microbes are normal inhabitants in biofilms in water, so the attention is less on eliminating these organisms from entering the system and more on managing their occurrence and concentrations in the pipe network. With anticipated warming trends associated with climate change, the factors that drive the growth of opportunistic pathogens in drinking water systems will likely increase. It is important, therefore, to evaluate treatment barriers and management activities for control of opportunistic pathogen risks. Controls for primary treatment, particularly for turbidity management and disinfection, should be reviewed to ensure adequacy for opportunistic pathogen control. However, the major focus for the utility’s opportunistic pathogen risk reduction plan is the management of biological activity and biofilms in the distribution system. Factors that influence the growth of microbes (primarily in biofilms) in the distribution system include, temperature, disinfectant type and concentration, nutrient levels (measured as AOC or BDOC), stagnation, flushing of pipes and cleaning of storage tank sediments, and corrosion control. Pressure management and distribution system integrity are also important to the microbial quality of water but are related more to the intrusion of contaminants into the distribution system rather than directly related to microbial growth. Summarizing the identified risk from drinking water, the availability and quality of disinfection data for treatment, and guidelines or standards for control showed that adequate information is best available for management of L. pneumophila. For L. pneumophila, the risk for this organism has been clearly established from drinking water, cases have increased worldwide, and it is one of the most identified causes of drinking water outbreaks. Water management best practices (e.g., maintenance of a disinfectant residual throughout the distribution system, flushing and cleaning of sediments in pipelines and storage tanks, among others) have been shown to be effective for control of L. pneumophila in water supplies. In addition, there are well documented management guidelines available for the control of the organism in drinking water distribution systems. By comparison, management of risks for Mycobacteria from water are less clear than for L. pneumophila. Treatment of M. avium is difficult due to its resistance to disinfection, the tendency to form clumps, and attachment to surfaces in biofilms. Additionally, there are no guidelines for management of M. avium in drinking water, and one risk assessment study suggested a low risk of infection. The role of tap water in the transmission of the other opportunistic pathogens is less clear and, in many cases, actions to manage L. pneumophila (e.g., maintenance of a disinfectant residual, flushing, cleaning of storage tanks, etc.) will also be beneficial in helping to manage these organisms as well.

Research – Biocontrol of multidrug resistant pathogens isolated from fish farms using silver nanoparticles combined with hydrogen peroxide insight to its modulatory effect

Nature

This study was divided into two parts. The first part involved the isolation, and detection of the prevalence and antimicrobial resistance profile of Aeromonas hydrophila, Pseudomonas aeruginosa, and Vibrio species from Nile tilapia fish and marine aquatic water. One hundred freshly dead Nile tilapia fish were collected from freshwater aquaculture fish farms located in Al-Abbassah district, Sharkia Governorate, and 100 samples of marine aquatic water were collected from fish farms in Port Said. The second part of the study focused on determining the in vitro inhibitory effect of dual-combination of AgNPs-H2O2 on bacterial growth and its down regulatory effect on crucial virulence factors using RT-PCR. The highest levels of A. hydrophila and P. aeruginosa were detected in 43%, and 34% of Nile tilapia fish samples, respectively. Meanwhile, the highest level of Vibrio species was found in 37% of marine water samples. Additionally, most of the isolated A. hydrophila, P. aeruginosa and Vibrio species exhibited a multi-drug resistance profile. The MIC and MBC results indicated a bactericidal effect of AgNPs-H2O2. Furthermore, a transcriptional modulation effect of AgNPs-H2O2 on the virulence-associated genes resulted in a significant down-regulation of aerA, exoU, and trh genes in A. hydrophila, P. aeruginosa, and Vibrio spp., respectively. The findings of this study suggest the effectiveness of AgNPs-H2O2 against drug resistant pathogens related to aquaculture.

Research – Surveillance and source tracking of foodborne pathogens in the vegetable production systems of India

Science Direct

Abstract

Fresh vegetables are prone to foodborne pathogen infection due to improper cultivation methods and poor post-harvest handling practices. This paper describes the prevalence of foodborne pathogens in three vegetables (tomato, brinjal, and lablab) cultivated by small-scale producers of India. We assessed the possible entry of pathogens into the vegetable production chain. Samples from soil, manure, irrigation water, harvesting bins, and workers’ hands were assessed as sources of possible pathogen entry. In addition, further sampling was done to determine prevalence of these pathogens in the produce from the agricultural fields and markets. Total plate counts, yeast and mold count, total coliforms like general microbial indices, and possible foodborne pathogens like SalmonellaEscherichia coli O157, Staphylococcus aureusPseudomonas aeruginosa, and Norovirus were assessed in all the samples by standard microbiological, biochemical, and molecular methods. The results revealed that among the three vegetables we studied, brinjal had the highest load of general microbial indicators. Moreover, the total coliforms count of market vegetables was higher than that of on-farm vegetables, indicating poor hygiene in post-harvest operations. The pathogen Pseudomonas aeruginosa was found in the market tomatoes (0.35 log10 cfu/g), and E. coli was detected in the market samples of brinjal (2.98–3.89 log10 cfu/g) and lablab (0.59–2.21 log10 cfu/g), thereby revealing fecal cross-contamination during pre or post-harvest operations. None of the vegetable samples were contaminated with E. coli O157:H7, Staphylococcus aureusSalmonella sp., and norovirus. However, the microbiological source tracking of primary production farms showed that E. coli O157:H7 was present in manure (9/27), soil (6/27), harvest bins (6/27), and irrigation water samples (3/27) of SSP farms. In addition, soil (33%), manure (11%), and harvest bins (11%) had Pseudomonas aeruginosa. In conclusion, our study suggests that adequate knowledge of good agricultural practices for small-scale producers to cultivate microbiologically safe vegetable production and proper handling strategies for vegetable vendors is crucial to mitigate foodborne pathogen outbreaks through fresh vegetables.

Research – Sweet and sour synergy: exploring the antibacterial and antibiofilm activity of acetic acid and vinegar combined with medical-grade honeys

Microbiology Society

ABSTRACT

, a combination of honey and vinegar, has been used as a remedy for wounds and infections in historical and traditional medical settings. While honey is now clinically used to treat infected wounds, this use of a complex, raw natural product (NP) mixture is unusual in modern western medicine. Research into the antimicrobial activity of NPs more usually focuses on finding a single active compound. The acetic acid in vinegar is known to have antibacterial activity at low concentrations and is in clinical use to treat burn wound infections. Here, we investigated the potential for synergistic activity of different compounds present in a complex ingredient used in historical medicine (vinegar) and in an ingredient mixture (). We conducted a systematic review to investigate published evidence for antimicrobial effects of vinegars against human pathogenic bacteria and fungi. No published studies have explicitly compared the activity of vinegar with that of a comparable concentration of acetic acid. We then characterized selected vinegars by HPLC and assessed the antibacterial and antibiofilm activity of the vinegars and acetic acid, alone and in combination with medical-grade honeys, against and . We found that some vinegars have antibacterial activity that exceeds that predicted by their acetic acid content alone, but that this depends on the bacterial species being investigated and the growth conditions (media type, planktonic vs. biofilm). Pomegranate vinegars may be particularly interesting candidates for further study. We also conclude that there is potential for acetic acid, and some vinegars, to show synergistic antibiofilm activity with manuka honey.

Research – Prevalence of Multidrug-Resistant Pseudomonas aeruginosa Isolated from Dairy Cattle, Milk, Environment, and Workers’ Hands

MDPI

Abstract

Pseudomonas aeruginosa is an opportunistic pathogen causing severe infection in animals and humans. This study aimed to determine the ecological distribution and prevalence of multidrug-resistant (MDR) P. aeruginosa isolated from dairy cattle, the environment, and workers’ hand swabs. Samples (n = 440) were collected from farms and households (n = 3, each). Rectal swabs, udder skin swabs, milk, workers’ hand swabs, feed, water, water sources, and beddings were collected. Samples were subjected to the bacterial identification of P. aeruginosa via 16S rRNA. Antimicrobial resistance (AMR) was detected either phenotypically using an antibiotic susceptibility test or genotypically with AMR resistance genes (ARGs) such as drfA, sul1, and ermB. P. aeruginosa was detected on dairy farms and households (10.3–57.5%, respectively), with an average of 23.2%. The resistance of dairy farm strains was observed against sulfamethoxazole, imipenem, cefepime, piperacillin–tazobactam, and gentamycin (100%, 72.7%, 72.7%, 68.8%, and 63.3%, respectively). Meanwhile, the resistance of household strains was observed against sulfamethoxazole, imipenem, amoxicillin, gentamicin, cefepime, and erythromycin by 91.3%, 82.6%, 75.4%, 75.4%, 68.1%, and 63.8%, respectively. The susceptibility of farm strains was detected against norfloxacin, ciprofloxacin, and levofloxacin (90.9%, 84.8%, and 72.7%, respectively). Meanwhile, the susceptibility of household strains was detected against ciprofloxacin, amikacin, and norfloxacin (100%, 84.1%, and 72.5%, respectively). About 81.4% of P. aeruginosa strains were MDR. ARGs (drfA, sul1, and ermB) were detected in farm strains (48.5%, 72.7%, and 24.4%, respectively) and household strains (50.7%, 72.5%, and 47.8%, respectively). Almost all P. aeruginosa had MAR over 0.2, indicating repeated application of antibiotics. P. aeruginosa prevalence was fivefold higher in households than on farms. MDR strains were higher amongst household strains than farm strains.

Research – Antimicrobial Activity of Essential Oils in Vapor Phase In Vitro and Its Application in Combination with Lactic Acid to Improve Chicken Breast Shelf Life

MDPI

Abstract

The effect of essential oils (EOs) incorporated in their vapor phase combined with lactic acid immersion pretreatment was studied on fresh refrigerated chicken breast shelf life. Among the several EOs assayed, the in vitro results obtained from the vapor diffusion test allowed mustard, oregano, and garlic EOs to be selected due to their higher antimicrobial activity. In addition, it was possible to determine the EO minimum inhibitory concentrations against Pseudomonas aeruginosa and Escherichia coli and to identify EO binary mixtures showing synergistic or additive effects. Based on the obtained results, a ternary mixture constituted by 0.073, 0.292, and 0.146 µL/mL of headspace of mustard, oregano, and garlic, respectively, was proposed for its application to chicken breasts. The ternary mixture inhibitory action was confirmed in vitro against P. aeruginosa and E. coli. Furthermore, the presence of numerous compounds with recognized antimicrobial and antioxidant activity was found in its volatile phase through gas chromatography. When applying an EO mixture in its vapor phase in combination with 1.0% v/v of lactic acid immersion pre-treatment on refrigerated chicken breast, a decrease in mesophilic microorganisms’ growth rate as well as in lipid oxidation was observed. Moreover, in a preliminary sensory test, the treated chicken breast was found to be acceptable to consumers and showed no significant differences compared to untreated chicken. In conclusion, the combined use of lactic acid immersion and EOs in their vapor phase was an effective alternative to increase chicken breast shelf life.

Research – Killing of a Multispecies Biofilm Using Gram-Negative and Gram-Positive Targeted Antibiotic Released from High Purity Calcium Sulfate Beads

MDPI

Abstract

Background: Multispecies biofilm orthopedic infections are more challenging to treat than mono-species infections. In this in-vitro study, we aimed to determine if a multispecies biofilm, consisting of Gram positive and negative species with different antibiotic susceptibilities could be treated more effectively using high purity antibiotic-loaded calcium sulfate beads (HP-ALCSB) containing vancomycin (VAN) and tobramycin (TOB) in combination than alone. Methods: Three sets of species pairs from bioluminescent strains of Pseudomonas aeruginosa (PA) and Staphylococcus aureus (SA) and clinical isolates, Enterococcus faecalis (EF) and Enterobacter cloacae were screened for compatibility. PA + EF developed intermixed biofilms with similar cell concentrations and so were grown on 316L stainless steel coupons for 72 h or as 24 h agar lawn biofilms and then treated with HP-ALCSBs with single or combination antibiotics and assessed by viable count or bioluminescence and light imaging to distinguish each species. Replica plating was used to assess viability. Results: The VAN + TOB bead significantly reduced the PA + EF biofilm CFU and reduced the concentration of surviving antibiotic tolerant variants by 50% compared to single antibiotics. Conclusions: The combination of Gram-negative and positive targeted antibiotics released from HP-ALCSBs may be more effective in treating multispecies biofilms than monotherapy alone.

Research – Pseudomonas Biofilms Pose Food Safety Threat by Aiding Listeria Survival in Processing Environments

Food Safety.Com

Pseudomonas biofilms can aid the survival of Listeria monocytogenes cells even after disinfection, according to a recent study. Although Pseudomonas are often overlooked as a food safety hazard due to being associated with food spoilage rather than with human foodborne illnesses, the study’s findings suggest that the bacteria may pose a direct threat to food safety in the processing environment.

Pseudomonas are the most commonly found bacteria in food processing environments due to characteristics such as a high growth rate at low temperatures, a high tolerance of antimicrobial agents, and biofilm formation. The bacteria have been of special interest as colonizers in food processing environments, as a food spoilage organism, and as protectors of foodborne pathogens.

Previous research has demonstrated the possibility of surviving bacteria exposed to sub-lethal concentrations of disinfectants being able to co-select for both disinfectant- and antibiotic-resistant properties. Several studies also indicate that interspecies interactions in a biofilm could serve as an accelerator for horizontal gene transfer, as well as facilitate adaptation to environmental conditions and the subsequent decreased susceptibility to antimicrobials. It has been suggested that multi-species, Pseudomonas-dominated biofilms could host and shelter pathogens like L. monocyotogenes.

In the study, researchers from the Norwegian University of Science and Technology isolated Pseudomonas samples from cleaned and disinfected surfaces in a salmon processing facility. A total of 186 isolates were screened for biofilm formation at 12 °C, and were graded as strong, medium, or weak biofilm producers. A high variation in biofilm formation was observed, with 12 percent rated as strong, 29 percent as medium, and 27 percent as weak biofilm producers, as well as 29 percent not producing a detectable biofilm.

Research – Bactericidal efficacy and mechanism of alkaline electrolyzed water co-action with acidic electrolyzed water on Pseudomonas aeruginosa biofilm

Wiley Online

Abstract

Here, the bactericidal efficacy and mechanism of action of the combination of alkaline electrolyzed water (AlEW) and acidic electrolyzed water (AcEW) on Pseudomonas aeruginosa biofilm were investigated, in comparison with single AcEW or AlEW treatment. The mature P. aeruginosa biofilm was formed at 120 h of cultivation. The number of bacteria in the P. aeruginosa biofilm decreased by 7.87 and 2.51 Lg CFU/mL after single AcEW and single AlEW treatment for 8 min, while the sterilization rate reached 92.2% and 33.1%, respectively. The group of AcEW (7 min) co-action with AlEW (1 min) had a P. aeruginosa sterilization rate of 85%. The AlEW (3 min) co-action with AcEW (5 min) treatment showed the best bactericidal efficacy. With this treatment, the sterilization rate achieved 94.4%, and the extracellular protein, polysaccharide, and DNA were degraded with the ratios of 70.8%, 77.7%, and 70.5%, respectively. Thus, AlEW exhibited a major washing effect, which can destroy the biofilm structure of extracellular polymers to a certain extent, while AcEW displayed a significant bactericidal effect. These results suggest that the AlEW co-action with AcEW treatment may be a promising candidate suitable for bacterial biofilm sterilization.

Research – Report of the Scientific Committee of the Spanish Agency for Food Safety and Nutrition (AESAN) on the prospection of biological ha-zards of interest in food safety in Spain (2)

AESAN

This report addresses the prospection of biological hazards for some types of food that may pose a risk to the population and that are not currently included in the official control programs in Spain.

It completes and updates the 2018 report by the Scientific Committee of the Spanish Agency for Food Safety and Nutrition (AESAN, 2018). A number of bacteria that are significant contributors to nosocomial infections due to the increase in the number of multi-resistant strains of Acinetobacter spp. ,Klebsiella pneumoniae and Pseudomonas aeruginosa are listed first.

It is also addressed the study of the prevalence and possible control of Bacillus cereus and Cronobacter spp. presence in cereal flours and others, the revision of Campylobacter jejuni and/or Campylobacter coli in meats other than poultry, as well as the study of Shigatoxin-producing Escherichia coli. These latter two biological agents are much better known from the food control perspective, although there are control measures for Campylobacter spp. in poultry meat and not in other types of meat such as beef or pork and in the case of E. coli, producers of Shiga toxins, the control of this particular type of pathogenic strains in food has not been specifically addressed either.

Finally, tick-borne viral encephalitis, which can be transmitted to humans through the consumption of raw milk or raw dairy products, has been indicated as a viral hazard. The prospective study shows the need to determine the prevalence of multi-resistant bacteria of Acinetobacter baumannii, K. pneumoniae and P. aeruginosa in foods in Spain, especially in ready-to-eat foods such as salads and fresh plant-based foods. This is especially important due to the lack of data on the prevalence of these bacteria in foods in Spain. However, food research is carried out in neighbouring countries.

It is also necessary to include C. jejuni and/or C. coli in the investigations of beef and pork, since the incidence of these foodborne pathogens in humans is not explained solely by the presence of these agents in poultry meat, being their presence in other animals for slaughter also evident. Similarly, outbreaks of Shiga toxin-producing E. coli have been reported in Spain over the last 25 years, which makes it advisable to control them in beef, raw milk and leafy vegetables.

With regard to Cronobacter spp. and B. cereus, the importance of these agents can be demonstrated given their survival in powdery materials such as flours of different origins, including cereals, although the reported outbreaks do not seem to indicate a high prevalence. As regards the only viral hazard mentioned, it should be noted that the wide dispersion of the ticks that can transmit this virus, together with the potential consumption of raw milk, makes it advisable to investigate it in raw milk products.

However, the study of the actual infective capacity of this virus is not easy to establish with simple analytical methods. With this last exception, research for controlling all these biological hazards in food is possible, with classical or advanced methodologies that are robust enough, available for each case.