Elucidating the Role of Biofilm-Forming Microbial Communities in Fermentative Biohydrogen Process: An Overview.

Amongst the biofuels described in the literature, biohydrogen has gained heightened attention over the past decade due to its remarkable properties. Biohydrogen is a renewable form of H2 that can be produced under ambient conditions and at a low cost from biomass residues. Innovative approaches are continuously being applied to overcome the low process yields and pave the way for its scalability. Since the process primarily depends on the biohydrogen-producing bacteria, there is a need to acquire in-depth knowledge about the ecology of the various assemblages participating in the process, establishing effective bioaugmentation methods. This work provides an overview of the biofilm-forming communities during H2 production by mixed cultures and the synergistic associations established by certain species during H2 production. The strategies that enhance the growth of biofilms within the H2 reactors are also discussed. A short section is also included, explaining techniques used for examining and studying these biofilm structures. The work concludes with some suggestions that could lead to breakthroughs in this area of research.

Death at the interface: Nanotechnology's challenging frontier against microbial surface colonization.

The emergence of antimicrobial-resistant bacterial strains has led to novel approaches for combating bacterial infections and surface contamination. More specifically, efforts in combining nanotechnology and biomimetics have led to the development of next-generation antimicrobial/antifouling nanomaterials. While nature-inspired nanoscale topographies are known for minimizing bacterial attachment through surface energy and physicochemical features, few studies have investigated the combined inhibitory effects of such features in combination with chemical alterations of these surfaces. Studies describing surface alterations, such as quaternary ammonium compounds (QACs), have also gained attention due to their broad spectrum of inhibitory activity against bacterial cells. Similarly, antimicrobial peptides (AMPs) have exhibited their capacity to reduce bacterial viability. To maximize the functionality of modified surfaces, the integration of patterned surfaces and functionalized exteriors, achieved through physical and chemical surface alterations, have recently been explored as viable alternatives. Nonetheless, these modifications are prone to challenges that can reduce their efficacy considerably in the long term. Their effectiveness against a wider array of microbial cells is still a subject of investigation. This review article will explore and discuss the emerging trends in biomimetics and other antimicrobials while raising possible concerns about their limitations and discussing future implications regarding their potential combined applications.

Cross-regional scale pollution of freshwater biofilms unveiled by antibiotic resistance genes.

A comprehensive global profile of the distribution of ARGs in freshwater biofilms is lacking. We utilized metagenomic approaches to reveal the diversity, abundance, transferability and hosts of ARGs in 96 freshwater biofilm samples from 38 sampling sites across four countries. The abundant ARGs were associated with bacitracin, multidrug, polymyxin macrolide-lincosamide-streptogramin (MLS) aminoglycoside, ß-lactam, chloramphenicol, sulfonamide and tetracycline resistance, consistent with the spectrum of antibiotics commonly used in human or veterinary medicine. As expected, the resistome in freshwater biofilm habitats was significantly influenced by geographical location and human footprint. Based on the co-occurrence pattern revealed by network analysis, mdtC, kdpE, and emrB were proposed as ARG indicators in freshwater biofilms that can be used to evaluate the abundance of 46 other co-occurring ARG subtypes quantitatively. Metagenomic assembly analysis revealed that the identified ARGs were hosted by more than 46 bacterial phyla, including various pathogens, which greatly expands the knowledge of resistome diversity in freshwater biofilms. Our study points to the central roles of biofilms in harbouring ARGs. The results could enhance understanding the distribution of ARGs in freshwater habitats, thereby strengthening the global environmental risk assessment and management of ARGs.

Exploring the optimization of aerobic food waste digestion efficiency through the engineering of functional biofilm Bio-carriers.

The possibility of breaking down cellulose-rich food waste through biofilm engineering was investigated. Six previously isolated strains from naturally degrading fruits and vegetables, screened for biofilm-forming ability and cellulolytic activity, were selected to enrich a biocarrier seeding microbial consortium. The food waste model used in this study was cabbage which was aerobically digested under repeated water rinsing and regular effluent drainage. The engineered biocarrier biofilm's functionality was evaluated by tracing microbial succession following metagenomic sequencing, quantitative PCR, scanning electron microscopy, and cellulolytic activity before and after the digestion processes. The engineered microbial consortium demonstrated superior biofilm-forming ability on biocarriers than the original microbial consortium and generally displayed a higher cellulolytic activity. The presented study provides one of the few studies of food waste aerobic digestion using engineered biofilms. Insights presented in this study could help further optimize aerobic food waste digestion.

Questioning the source of identified non-foodborne pathogens from food-contact wooden surfaces used in Hong Kong's urban wet markets.

In this study, a phylogenic analysis was performed on pathogens previously identified in Hong Kong wet markets' cutting boards. Phylogenetic comparisons were made between phylotypes obtained in this study and environmental and clinical phylotypes for establishing the possible origin of selected bacterial species isolated from wet market cutting board ecosystems. The results reveal a strong relationship between wet market bacterial assemblages and environmental and clinically relevant phylotypes. However, our poor knowledge of potential cross-contamination sources within these wet markets is further exacerbated by failing to determine the exact or presumed origin of its identified pathogens. In this study, several clinically relevant bacterial pathogens such as Klebsiella pneumoniae, Streptococcus suis and Streptococcus porcinus were linked to cutting boards associated with pork; Campylobacter fetus, Staphylococcus aureus, Escherichia coli, and A. caviae in those associated with poultry; and Streptococcus varanii, A. caviae, Vibrio fluvialis, and Vibrio parahaemolyticus in those associated with seafood. Identifying non-foodborne clinically relevant pathogens in wet market cutting boards in this study confirms the need for safety approaches for wet market meat, including cold storage. The presented study justifies the need for future systematic epidemiological studies to determine identified microbial pathogens. Such studies should bring about significant improvements in the management of hygienic practices in Hong Kong's wet markets and work towards a One Health goal by recognizing the importance of wet markets as areas interconnecting food processing with animal and clinical environments.

Active Microbiome Structure and Functional Analyses of Freshwater Benthic Biofilm Samples Influenced by RNA Extraction Methods.

Advances in high-throughput sequencing technologies have enabled extensive studies of freshwater biofilms and significant breakthroughs in biofilm meta-omics. To date, however, no standardized protocols have been developed for the effective isolation of RNA from freshwater benthic biofilms. In this study, we compared column-based kit RNA extraction with five RNAzol-based extractions, differentiated by various protocol modifications. The RNA products were then evaluated to determine their integrity, purity and yield and were subjected to meta-transcriptomic sequencing and analysis. Significant discrepancies in the relative abundance of active communities and structures of eukaryotic, bacterial, archaebacterial, and viral communities were observed as direct outcomes of the tested RNA extraction methods. The column isolation-based group was characterized by the highest relative abundance of Archaea and Eukaryota, while the organic isolation-based groups commonly had the highest relative abundances of Prokaryota (bacteria). Kit extraction methods provided the best outcomes in terms of high-quality RNA yield and integrity. However, these methods were deemed questionable for studies of active bacterial communities and may contribute a significant degree of bias to the interpretation of downstream meta-transcriptomic analyses.

Biomimetic Polymer Surfaces by High Resolution Molding of the Wings of Different Cicadas.

Recent studies have shown that insect wings have evolved to have micro- and nanoscale structures on the wing surface, and biomimetic research aims to transfer such structures to application-specific materials. Herein, we describe a simple and cost-effective method of replica molding the wing topographies of four cicada species using UV-curable polymers. Different polymer blends of polyethylene glycol diacrylate and polypropylene glycol diacrylate were used as molding materials and a molding chamber was designed to precisely control the x, y, and z dimensions. Analysis by scanning electron microscopy showed that structures ranged from 148 to 854 nm in diameter, with a height range of 191-2368 nm, and wing patterns were transferred with high fidelity to the crosslinked polymer. Finally, bacterial cell studies show that the wing replicas possess the same antibacterial effect as the cicada wing from which they were molded. Overall, this work shows a quick and simple method for patterning UV-curable polymers without the use of expensive equipment, making it a highly accessible means of producing microstructured materials with biological properties.

Ecotoxicological effects of erythromycin on a multispecies biofilm model, revealed by metagenomic and metabolomic approaches.

The presence of antibiotics such as erythromycin, even in trace amounts, has long been acknowledged for negatively impacting ecosystems in freshwater environments. Although many studies have focused on the impact of antibiotic pollution at a macroecological level, the impact of erythromycin on microecosystems, such as freshwater biofilms, is still not fully understood. This knowledge gap may be attributed to the lack of robust multispecies biofilm models for fundamental investigations. Here, we used a lab-cultured multispecies biofilm model to elucidate the holistic response of a microbial community to erythromycin exposure using metagenomic and metabolomic approaches. Metagenomic analyses revealed that biofilm microbial diversity did not alter following erythromycin exposure. Notably, certain predicted metabolic pathways such as cell-cell communication pathways, amino acid metabolism, and peptidoglycan biosynthesis, mainly by the phyla Actinobacteria, Alpha/Beta-proteobacteria, Bacteroidetes, and Verrucomicrobia, were found to be involved in the maintenance of homeostasis-like balance in the freshwater biofilm. Further untargeted metabolomics data highlighted changes in lipid metabolism and linoleic acid metabolism and their related molecules as a direct consequence of erythromycin exposure. Overall, the study presented a unique picture of how multispecies biofilms respond to single environmental stress exposures. Moreover, the study demonstrated the feasibility of using lab simulated multispecies biofilms for investigating their interaction and reactivity of specific bioactive compounds or pollutants at a fundamental level.

The Potential of Single-Cell Oils Derived From Filamentous Fungi as Alternative Feedstock Sources for Biodiesel Production.

Microbial lipids, also known as single-cell oils (SCOs), are highly attractive feedstocks for biodiesel production due to their fast production rates, minimal labor requirements, independence from seasonal and climatic changes, and ease of scale-up for industrial processing. Among the SCO producers, the less explored filamentous fungi (molds) exhibit desirable features such as a repertoire of hydrolyzing enzymes and a unique pellet morphology that facilitates downstream harvesting. Although several oleaginous filamentous fungi have been identified and explored for SCO production, high production costs and technical difficulties still make the process less attractive compared to conventional lipid sources for biodiesel production. This review aims to highlight the ability of filamentous fungi to hydrolyze various organic wastes for SCO production and explore current strategies to enhance the efficiency and cost-effectiveness of the SCO production and recovery process. The review also highlights the mechanisms and components governing lipogenic pathways, which can inform the rational designs of processing conditions and metabolic engineering efforts for increasing the quality and accumulation of lipids in filamentous fungi. Furthermore, we describe other process integration strategies such as the co-production with hydrogen using advanced fermentation processes as a step toward a biorefinery process. These innovative approaches allow for integrating upstream and downstream processing units, thus resulting in an efficient and cost-effective method of simultaneous SCO production and utilization for biodiesel production.

Impacts of Wet Market Modernization Levels and Hygiene Practices on the Microbiome and Microbial Safety of Wooden Cutting Boards in Hong Kong.

Accessing food through wet markets is a common global daily occurrence, where fresh meat can be purchased to support an urbanizing world population. Similar to the wet markets in many other metropolitan cities in Asia, Hong Kong wet markets vary and are characterized by differing hygiene routines and access to essential modern technologies. The lack of risk assessments of food contact surfaces in these markets has led to substantial gaps in food safety knowledge and information that could help improve and maintain public health. Microbial profiling analyses were conducted on cutting boards that had been used to process pork, poultry, and seafood at 11 different wet markets. The markets differed in hygiene protocols and access to modern facilities. Irrespective of whether wet markets have access of modern infrastructure, the hygiene practices were largely found to be inefficient based on the prevalence of bacterial species typically associated with foodborne pathogens such as Campylobacter fetus, Clostridium perfringens, Staphylococcus aureus, and Vibrio parahaemolyticus; indicator organisms such as Escherichia coli; as well as nonfoodborne pathogenic bacterial species potentially associated with nosocomial infections, such as Klebsiella pneumoniae and Enterobacter cloacae. Other Vibrio species, V. parahaemolyticus and V. vulnificus, typically associated with contaminated raw or undercooked seafood with the potential to cause illness in humans, were also found on wooden cutting boards. This study indicated that the hygienic practices used in Hong Kong wet markets are not sufficient for preventing the establishment of spoilage or pathogenic organisms. This study serves as a basis to review current hygiene practices in wet markets and provides a framework to reassess existing safety protocols.

Effects of sodium citrate on the structure and microbial community composition of an early-stage multispecies biofilm model.

In recent years, most biofilm studies have focused on fundamental investigations using multispecies biofilm models developed preferentially in simulated naturally occurring low-nutrient medium than in artificial nutrient-rich medium. Because biofilm development under low-nutrient growth media is slow, natural media are often supplemented with an additional carbon source to increase the rate of biofilm formation. However, there are knowledge gaps in interpreting the effects of such supplementation on the resulting biofilm in terms of structure and microbial community composition. We investigated the effects of supplementation of a simulated freshwater medium with sodium citrate on the resulting structure, bacterial community composition, and microbial network interactions of an early-stage multispecies biofilm model. Qualitative and quantitative analyses of acquired confocal laser scanning microscopy data confirmed that sodium citrate supplementation distinctly increased biofilm biomass. Sequencing data revealed that the microbial community structure of biofilms grown in sodium citrate-supplemented conditions was characterized with increased relative abundance and dominance of Proteobacteria compared with that of biofilms grown in sodium citrate-free conditions. Our findings suggest that the supplementation of a low-nutrient medium with a carbon source in experiments involving multispecies biofilms may lead to structural and compositional biases of the microbial community, causing changes in biofilm phenotype.

Identifying Breast Cancer Care Quality Measures for a Cancer Facility in Rural Sub-Saharan Africa: Results of a Systematic Literature Review and Modified Delphi Process.

PURPOSE: The burden of cancer is growing in low- and middle-income countries (LMICs), including sub-Saharan Africa. Ensuring the delivery of high-quality cancer care in such regions is a pressing concern. There is a need for strategies to identify meaningful and relevant quality measures that are applicable to and usable for quality measurement and improvement in resource-constrained settings. METHODS: To identify quality measures for breast cancer care at Butaro Cancer Center of Excellence (BCCOE) in Rwanda, we used a modified Delphi process engaging two panels of experts, one with expertise in breast cancer evidence and measures used in high-income countries and one with expertise in cancer care delivery in Rwanda. RESULTS: Our systematic review of the literature yielded no publications describing breast cancer quality measures developed in a low-income country, but it did provide 40 quality measures, which we adapted for relevance to our setting. After two surveys, one conference call, and one in-person meeting, 17 measures were identified as relevant to pathology, staging and treatment planning, surgery, chemotherapy, endocrine therapy, palliative care, and retention in care. Successes of the process included participation by a diverse set of global experts and engagement of the BCCOE community in quality measurement and improvement. Anticipated challenges include the need to continually refine these measures as resources, protocols, and measurement capacity rapidly evolve in Rwanda. CONCLUSION: A modified Delphi process engaging both global and local expertise was a promising strategy to identify quality measures for breast cancer in Rwanda. The process and resulting measures may also be relevant for other LMIC cancer facilities. Next steps include validation of these measures in a retrospective cohort of patients with breast cancer.

Insights into the Microbiological Safety of Wooden Cutting Boards Used for Meat Processing in Hong Kong's Wet Markets: A Focus on Food-Contact Surfaces, Cross-Contamination and the Efficacy of Traditional Hygiene Practices.

Hong Kong's wet markets play a crucial role in the country's supply of safe, fresh meat to satisfy the dietary needs of its population. Whilst food safety regulations have been introduced over the past few years to maintain the microbial safety of foods sold from these wet markets, it remains unclear whether the hygiene maintenance that is performed on the wooden cutting boards used for meat-processing is effective. In fact, hygiene maintenance may often be overlooked, and hygiene standards may be insufficient. If so, this may lead to the spread of harmful pathogens through cross-contamination, thereby causing severe risks to public health. The aim of this study was to determine the level of microbial transfer between wooden cutting boards and swine meat of various qualities, using 16S metagenomic sequencing, strain identification and biofilm screening of isolated strains. The results established that: (a) the traditional hygiene practices used for cleaning wooden cutting boards in Hong Kong's wet markets expose the surfaces to potentially harmful microorganisms; (b) the processing of microbially contaminated meat on cutting boards cleaned using traditional practices leads to cross-contamination; and (c) several potentially pathogenic microorganisms found on the cutting boards have good biofilm-forming abilities. These results reinforce the need to review the traditional methods used to clean wooden cutting boards after the processing of raw meat in Hong Kong' wet markets so as to prevent cross-contamination events. The establishment of proper hygiene protocols may reduce the spread of disease-causing microorganisms (including antibiotic-resistant microorganisms) in food-processing environments.

Large-Scale Screening of 239 Traditional Chinese Medicinal Plant Extracts for Their Antibacterial Activities against Multidrug-Resistant Staphylococcus aureus and Cytotoxic Activities.

Novel alternative antibacterial compounds have been persistently explored from plants as natural sources to overcome antibiotic resistance leading to serious foodborne bacterial illnesses. In this study, the ethanolic extracts from 239 traditional Chinese medicinal plants (TCMP)' materials were screened to discover promising candidates that have strong antibacterial properties against multidrug-resistant Staphylococcus (S.) aureus and low cytotoxicity. The results revealed that 74 extracts exhibited good antibacterial activities (diameter of inhibition zone (DIZ) ≥ 15 mm). Furthermore, 18 extracts (DIZ ≥ 20 mm) were determined their minimum inhibitory concentrations (MIC) and minimum bactericide concentrations (MBC), ranging from 0.1 to 12.5 mg/mL and 0.78 to 25 mg/mL, respectively. In addition, most of the 18 extracts showed relatively low cytotoxicity (a median lethal concentration (LC50) >100 µg/mL). The 18 extracts were further determined to estimate possible correlation of their phenolic contents with antibacterial activity, and the results did not show any significant correlation. In conclusion, this study selected out some promising antibacterial TCMP extracts with low cytotoxicity, including Rhus chinensis Mill., Ilex rotunda Thunb., Leontice kiangnanensis P.L.Chiu, Oroxylum indicum Vent., Isatis tinctorial L., Terminalia chebula Retz., Acacia catechu (L.f.) Willd., Spatholobus suberectus Dunn, Rabdosia rubescens (Hemsl.) H.Hara, Salvia miltiorrhiza Bunge, Fraxinus fallax Lingelsh, Coptis chinensis Franch., Agrimonia Pilosa Ledeb., and Phellodendron chinense C.K.Schneid.

A Field Study Into Hong Kong's Wet Markets: Raised Questions Into the Hygienic Maintenance of Meat Contact Surfaces and the Dissemination of Microorganisms Associated With Nosocomial Infections.

Millions every day purchase their raw meat in wet markets around the globe, especially in Hong Kong city, where modern and a traditional way of living is made possible. While food hygiene standards in Hong Kong have more recently focused on the safety of meat sold in these wet markets, the hygienic surface level of wooden cutting boards used for processing meats is seldom observed. This original study performed microbial community profiling, as well as isolating and identifying various strains multiple wooden cutting boards from nine wet markets located on Hong Kong Island. Our study also investigated the efficiency of scraping the surface of cutting boards as a traditional cleaning technique in Hong Kong. Results indicate that these hygienic practices are inefficient for guarantying proper surface hygiene as some most tested cutting boards were found to harbor microbial species typically associated with hospital nosocomial infections, such as Klebsiella pneumoniae. Further analysis also led to discovering the presence of antibiotic-resistant genes (ARGs) among isolated strains. Our results showcase the significance and effects of cross-contamination in Hong Kong wet markets, especially with regards to the potential spreading of clinically-relevant strains and ARGs on food processing surfaces. This study should, therefore, serve as a basis to review current hygienic practices in Hong Kong's wet market on a larger scale, thereby improving food safety and ultimately, public health.

Standardization of Education of Patients With Cancer in a Low- and Middle-Income Country: A Quality Improvement Project Using the Cancer and You Booklet.

PURPOSE: The Butaro Cancer Centre of Excellence is the first comprehensive referral cancer center in Rwanda and at its inception did not have a standardized patient education program. Partners in Health/Inshuti Mu Buzima and the Rwandan Ministry of Health conducted a quality improvement project to increase patient knowledge by implementing a standardized oncology education program using picture-based and culturally appropriate materials designed for patients with cancer in low- and middle-income countries. METHODS: Four Rwandan nurses were trained to provide patient education using the Cancer and You education booklet created by Global Oncology. A pre- and post-test design was used to evaluate patients' knowledge of cancer, treatment, and management of adverse effects. Nurses administered a posteducation questionnaire in Kinyarwanda to determine patients' level of satisfaction with the education session and booklet. The four nurses were interviewed at the completion of the project for their feedback. A total of 40 oncology patients were included in the pilot project, of which 85% reported completing primary school or less. RESULTS: On average, participants improved 19% (95% CI, 13.9% to 24.1%; standard deviation, 16%) from pre- to postevaluation, demonstrating a significant increase in knowledge (P ≤ .001). Nearly all patients (97.5%) reported that they were either satisfied or very satisfied with the education program. Oncology nurses gave positive feedback, highlighting that it was helpful to have a standard tool for education with descriptive illustrations for those patients with low literacy. CONCLUSION: Implementation of a standardized patient education program demonstrated a statistically significant increase in patient knowledge and a high level of satisfaction among patients and nurses. The project serves as an example for other low- and middle-income countries looking to standardize oncology patient education.

Could benthic biofilm analyses be used as a reliable proxy for freshwater environmental health?

The quality of freshwater undoubtedly reflects the health of our surrounding environment, society, and economy, as these are supported by various freshwater ecosystems. Monitoring efforts have therefore been considered a vital means of ensuring the ecological health of freshwater environments. Nevertheless, most aquatic environmental monitoring strategies largely focus on bulk water sampling for analysis of physicochemical and key biological indicators, which for the most part do not consider pollution events that occur at any time between sampling events. Because benthic biofilms are ubiquitous in aquatic environments, pollution released during sporadic events may be absorbed by these biofilms, which can act as repositories of pollutants. The aim of this study was to assess whether benthic biofilm monitoring could provide an efficient way of properly characterizing the extent of pollution in aquatic environments. Here, bulk water and benthic biofilms were sampled from three Hong Kong streams having various pollution profiles, and subsequently compared via high-resolution microscopy, metagenomic analysis, and analytical chemistry. The results indicated that biofilms were, indeed, reservoirs of environmental pollutants, having different profiles compared with that of the corresponding bulk water samples. Moreover, the results also suggested that biofilms sampled in polluted areas were characterized by a higher species richness. While the analytical testing of benthic biofilms still needs further development, the integration of chemical-pollutant profiles and biofilm sequencing data in future studies may provide unique perspectives for understanding and identifying pollution-related biofilm biomarkers.

Effects of Lactic Acid Bacteria-Fermented Soymilk on Isoflavone Metabolites and Short-Chain Fatty Acids Excretion and Their Modulating Effects on Gut Microbiota.

Lactobacillus rhamnosus strain ASCC 1520 with high soy isoflavone transformation ability was used to ferment soymilk and added to the diet of mice. The impact of L. rhamnosus fermentation on soy isoflavone metabolites and intestinal bacterial community, in conjunction with fecal enzyme activity and short-chain fatty acids (SCFA) excretion was evaluated. Antibiotics intervention resulted in a decrease in fecal enzyme activities and SCFA. Although long-term intake of soymilk or L. rhamnosus-fermented soymilk did not affect the fecal ß-glucuronidase and ß-galactosidase activities, it improved the ß-glucosidase activity when antibiotics were concomitantly administered. Soymilk or fermented soymilk administration increased the isoflavone metabolites (O-DMA and equol) excreted in urine. Antibiotics decreased the daidzein excretion and its metabolites but showed little effect on glycitein and genistein excretion. Principal coordinates analysis (PCoA) of the 16s rRNA gene sequencing data found a remarkable shift in gut microbiota after soymilk administration and antibiotics treatment. Matastats test of the relative abundance of bacterial taxa revealed Odoribacter (Bacteroidales family), Lactobacillus (Lactobacillales order), and Alistipes (Rikenellaceae family) were enriched in soymilk while bacterial taxa from Bacteroides and Lactobacillus were enriched in L. rhamnosus-fermented soymilk. Furthermore, there was less decrease in bacterial taxa with fermented soymilk group even when antibiotics were concomitantly administered. Overall, this study revealed that the gut microbiota of a healthy host is enough for the whole isoflavone metabolism under normal conditions. Feeding mice with L. rhamnosus-fermented soymilk improved fecal enzyme activity and kept the balance of the gut mirobiota when antibiotics were used. PRACTICAL APPLICATION: Feeding mice with L. rhamnosus-fermented soymilk improved fecal enzyme activity and kept the balance of the gut mirobiota when antibiotics were used.

Biofilm research within irrigation water distribution systems: Trends, knowledge gaps, and future perspectives.

Biofilms in irrigation water distribution systems (IWDSs) play an essential role in spreading pathogens, chemical pollutants, and environmental pollutants into downstream irrigated crops and thus should be considered a potential threat to food safety. Although the role of biofilms in drinking water distribution systems has been extensively studied in the last decade, the research on IWDS biofilms in this period has been limited. This review identifies research gaps in the field of IWDS biofilms, provides perspectives on experimental designs for investigating IWDS biofilms, and suggests potential strategies worth pursuing in IWDS management. The current state of the art of IWDS biofilms is discussed, and an analysis of the challenges in IWDS biofilm research is presented. Furthermore, this review proposes useful advanced technologies that allow a practical, in-depth fundamental understanding of IWDS biofilms. In a nutshell, this article provides future directions and insights into detailed experimental designs on a relatively under-reported research topic: "IWDS biofilms."