Microbial risk assessment of Staphylococcus species in smoked fish from informal markets in Edo State, Nigeria: processing practices and public health implications

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Boyo-Agbonile, Abraham G. Ogofure, Brenda O. Isichei-Ukah, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8482256/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 01 May, 2026 Read the published version in Annals of Microbiology → Version 1 posted 4 You are reading this latest preprint version Abstract Purpose Staphylococcal contamination of smoked fish represents a public health concern, particularly in informal food markets. This study investigated the microbial risk associated with Staphylococcus -contaminated smoked fish, processing practices, and potential public health implications in Benin City, Nigeria. Structured questionnaires were administered to smoked fish vendors to document processing, handling, and storage practices. Materials and Methods Smoked fish samples (Scomber, Meluza, Sese, catfish, and herring) were collected and analyzed using standard culture-based and molecular techniques. Questionnaire data were analyzed using chi-square and correlation analyses. Quantitative Microbial Risk Assessment (QMRA) was conducted to estimate the probability of adverse health outcomes following consumption of contaminated fish, using the beta-Poisson dose–response model. Results The results showed that 98% of the smoked fish samples were contaminated with Staphylococcus species, including coagulase-positive strains. Methicillin-resistant Staphylococcus aureus (MRSA) was detected in 37.78% of the samples. Fish stored for more than two days post-smoking exhibited significantly higher bacterial counts ( p < 0.05), with marked logarithmic increases observed within 24–48 h after smoking. Scomber and Meluza samples had significantly higher staphylococcal loads compared with other fish species. The use of preservatives, vendors’ awareness of bacterial contamination, and the type of smoking material (wood or sawdust) did not significantly ( p > 0.05) influence bacterial loads. QMRA outcomes indicated a high predicted probability (> 90%) of adverse health effects under ambient storage conditions over 2–4 d. Conclusion These findings underscore the need for improved hygiene practices, enhanced consumer and vendor education, better preservation strategies, and stricter regulatory oversight to reduce the risk of staphylococcal foodborne illnesses associated with smoked fish consumption. Staphylococcus spp. Quantitative Microbial Risk Assessment (QMRA) Beta-Poisson model Smoked fish Food safety Processing practices MRSA Figures Figure 1 Figure 2 Figure 3 Introduction Fish is a critical source of high-quality protein, micronutrients, and essential fatty acids, making it a dietary staple for millions of people globally (Obiero et al., 2019 ; Mohanty et al., 2019 ; Aremu et al., 2021 ; Ekonomou et al., 2022 ), particularly in regions like Nigeria. However, postharvest losses due to microbial spoilage are a significant concern, especially in developing countries where traditional fish preservation methods dominate (Ghaly, 2010 ; Diop et al., 2016 ; Aremu et al., 2021 ). Smoking is one of the most common preservation techniques in these regions, utilized not only to enhance flavour and texture but also to prolong shelf life by reducing moisture content and inhibiting microbial growth (Shajil et al., 2018 ; Olusola, 2021 ; Ekonomou et al., 2022 ; Deliephan et al., 2023 ). While this method has proven to be effective against some spoilage organisms, it may also create an environment conducive to contamination by pathogens such as Staphylococcus aureus , posing a substantial risk to public health (Ghaned et al., 2019 ; Musa et al., 2020 ; Doutoum et al., 2023 ). Staphylococcus aureus is a facultative anaerobic bacterium and opportunistic pathogen that thrives in various environmental conditions and is commonly associated with foodborne illnesses (Ogofure & Igbinosa, 2021 ; Merghni et al., 2023 ). The bacterium produces a wide range of virulence factors, including heat-stable enterotoxins, which can persist in smoked fish even after processing (Puah et al., 2016 ; Fisher et al., 2018 ). In developing countries, the challenges of ensuring hygienic food handling practices are exacerbated by limited regulatory oversight, inadequate infrastructure, and the widespread use of artisanal processing methods (Alimi, 2016 ; Morse et al., 2018 ; Abdi et al., 2020 ). As such, smoked fish sold in open markets across Nigeria may become a reservoir for pathogenic bacteria, including coagulase-positive Staphylococcus species like S. aureus , whose heat-stable toxin can remain viable under cooking or cells can survive suboptimal cooking or processing temperatures. Consumption of smoked fish contaminated with S. aureus can pose a significant threat to public health, as the ingestion of staphylococcal enterotoxins can lead to staphylococcal food poisoning (SFP), characterized by nausea, vomiting, abdominal cramps, and diarrhea (Wu et al., 2016 ; Schelin et al., 2017 ; Guo et al., 2023 ). Given the widespread reliance on smoked fish as a protein source in Nigeria, understanding the extent of microbial contamination and the associated risk to consumers is imperative. In addition to enterotoxin production, antibiotic-resistant strains of S. aureus , particularly methicillin-resistant S. aureus (MRSA), are becoming an increasing concern globally (Liu et al., 2021 ; Nandhini et al., 2022 ). These strains complicate treatment options for foodborne illness and contribute to the growing burden of antimicrobial resistance (AMR), a public health crisis recognized by the World Health Organization (WHO) (Liu et al., 2021 ). In Benin City, Nigeria, limited access to refrigeration and reliance on artisanal preservation practices heighten the risk of bacterial contamination in smoked fish, a dietary staple. Thus, while smoked fish is widely consumed, there is limited information on the prevalence of S. aureus in our study location. Furthermore, the identification of the specific processing practices that may contribute to bacterial contamination, and evaluation of the probability of adverse health outcome based on consumption quantity or patterns will be crucial to assess the probable risk of S. aureus in smoke fish staple. This study addresses a critical gap in the literature by conducting a comprehensive microbial risk assessment of S. aureus -contaminated smoked fish sold in Benin City, Nigeria, and the hygienic conditions (processing practices) from the vendor's perspective that may impact consumer health. Therefore, we aim to provide a detailed assessment of fish processing practices and the associated microbial risks with emphasis on Staphylococcus species contamination from fish samples, specifically looking at Coagulase positive Staphylococcus aureus , methicillin resistant Staphylococcus aureus and coagulase negative staphylococci. Additionally, our study examines the public health implications, particularly in the context of food safety regulations. By integrating microbial risk assessment models with an analysis of vendor practices, this work aims to inform policy development and promote safer food processing methods, ultimately reducing the burden of foodborne illness and supporting sustainable food systems in Nigeria and beyond. Methodology Study Design and Location This study was conducted across twenty (20) open markets located in three Local Government Areas (LGA) within the Edo South Senatorial district of Benin City, Nigeria. Smoked fish samples were collected from these markets and categorized according to local terminology into different groups, which include Scomber or Titus fish ( Scomber scombrus ), Hake or Meluza ( Merluccius merluccius ), Catfish ( Clarias gariepinus ), Sese or Horse Mackerel ( Trachurus trachurus ) and Herring or Shawa ( Clupea harengus ). Heterotrophic bacterial counts were determined, and staphylococci isolates were differentiated into coagulase-negative, coagulase-positive, methicillin-resistant Staphylococcus aureus (MRSA), and non-MRSA groups. The prevalence of pathogenic staphylococcal isolates was also evaluated. In addition, a structured questionnaire was administered to vendors to assess postharvest handling practices and hygiene-related behaviors associated with smoked fish processing. The questionnaire is provided in the Supplementary Material (S1.0). All questionnaire responses were anonymized, and no personally identifiable information was collected. Sampling Strategy In this study, a multi-stage sampling approach was employed covering three Local Government Areas (LGAs) in the Edo South Senatorial district, which were purposively selected based on high fish trading activity and large population density. Within each LGA, open markets were randomly selected from an official list of markets, yielding a total of twenty markets. Within each market, vendors were approached at random until the target sample size was achieved. A total of 405 vendors were approached, of whom 330 agreed to participate and provided written informed consent, resulting in a response rate of 81.48%. Vendors who declined participation cited time constraints or privacy concerns and were excluded from the study. Each participating vendor provided one smoked fish sample, resulting in a total of 330 fish samples collected for microbiological analysis. Questionnaire Design, Administration, and Data Processing A structured questionnaire was designed and developed based on existing literature on food safety practices (Teffo & Tabit, 2020 ; Halabi et al., 2023 ; Alqadiri et al., 2025 ) and refined through expert consultation with food microbiologists and public health specialists in the Microbiology Postgraduate Board, University of Benin (during proposal presentation). The questionnaire was designed to assess postharvest handling practices, hygienic behaviors and conditions, and vendor awareness regarding bacterial contamination of smoked fish. The questionnaire comprised four key construct domains which include (1) demographic variables, to evaluate socioeconomic factors influencing hygiene practices; (2) processing practices, to assess potential risk factors associated with product handling; (3) preservation and storage, to identify critical control points related to product safety; and lastly (4) hygiene awareness, a recognized predictor of food safety behaviors (Abdi et al., 2020 ; Teffo & Tabit, 2020 ; Alqadiri et al., 2025 ). Prior to data collection, the questionnaire was pre-tested with five smoked fish vendors from a market outside the study area to assess clarity, relevance, and contextual appropriateness. Feedback from the pilot testing informed minor revisions before final deployment. The finalized questionnaire (Supplementary Material S1.0) was administered in person by trained research assistants fluent in English and the local dialect (Bini), enabling participation by vendors with limited literacy. Questionnaires were administered at vendors’ points of sale after participants had received an explanation of the study objectives and procedures and had provided written informed consent. A total of 330 vendors participated in the survey, corresponding to the number of smoked fish samples collected. However, some questionnaires contained incomplete or internally inconsistent responses for one or more variables. To ensure analytical validity, only questionnaires with complete responses for the relevant variables were included in the final descriptive and inferential analyses (complete-case analysis), resulting in an effective questionnaire sample size of 301 respondents. All microbiological analyses were conducted on the full set of 330 smoked fish samples. Completed questionnaires were anonymized at the point of collection, coded, and entered into Microsoft Excel for data cleaning prior to statistical analysis Sample Collection Using a prevalence rate of 26%, as observed in similar studies from Nigeria (Vazquez-Sanchez et al., 2012 ), the sample size was determined to ensure adequate statistical power. The sample size and parameters were determined using the formula. $$\:Sample\:size\:\left(N\right)=\:{(Z}^{2\:}P\:\left(1-P\right))÷{D}^{2}\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:\:(1)$$ where: N = Sample size, Z = z-score (1.969), P = estimated prevalence (0.26) and D = 0.05 (margin of error). Based on this calculation, a minimum sample size of 299 was obtained. This value was increased to 330 to account for potential non-response and incomplete data. The weight of each fish sample was recorded, and visual inspection of the smoked fish was conducted prior to microbiological analysis. Staphylococcal Culture and Enumeration Twenty-five grams (25 g) of each smoked fish sample were aseptically weighed and homogenized separately in two different sterile diluents to support differential microbial enumeration. For total heterotrophic bacterial counts, samples were homogenized in 225 mL of sterile physiological saline (0.85% w/v NaCl). In parallel, a separate homogenate of 25 g of the same sample was prepared in 225 mL of sterile high-salt saline (7.5% w/v NaCl) to favour the recovery of halotolerant staphylococci while suppressing the growth of non-halotolerant background microbiota. Each homogenate was thoroughly mixed, and tenfold serial dilutions were prepared using the corresponding saline diluent. From the 0.85% saline series, 1.0 mL aliquots from appropriate dilutions (typically 10 ⁻³ ) were plated onto nutrient agar using the pour-plate method for total heterotrophic bacterial enumeration. From the 7.5% saline series, 1.0 mL aliquots from lower dilutions (typically 10 ⁻1 ) were plated onto Mannitol Salt Agar (MSA) for the selective enumeration of presumptive staphylococci. All inoculated plates were incubated aerobically at 37°C for 24 h. Following incubation, colonies were counted on plates containing 30–300 colonies and results were expressed as colony-forming units per gram (CFU/g) of sample. On MSA, presumptive staphylococci were identified based on their ability to grow under high-salt conditions and by mannitol fermentation reactions, indicated by yellow (mannitol-positive) or pink (mannitol-negative) colonies. Representative colonies exhibiting distinct morphologies were subcultured onto fresh nutrient agar to obtain pure isolates for subsequent phenotypic (Bridson, 2006 ; Ogofure & Igbinosa, 2021 ). All experiments were conducted in three independent biological replicates. Based on the lowest dilution plated and a plated volume of 1.0 mL, the detection limit of the enumeration procedure was 10 CFU/g (1.0 log₁₀ CFU/g). Characterization of Staphylococcal Isolates from Smoked Fish Presumptive staphylococcal isolates initially enumerated on Mannitol Salt Agar were further characterized phenotypically. Coagulase-positive staphylococci were identified using Vogel-Johnson Agar (Oxoid), supplemented with 1% potassium tellurite solution, which allows for differentiation based on tellurite reduction and mannitol fermentation. For the detection of methicillin-resistant Staphylococcus aureus (MRSA), isolates were cultured on Oxacillin Resistance Screening Agar containing oxacillin supplement, following standard guidelines (Bridson, 2006 ; Ogofure & Igbinosa, 2021 ). Phenotypic identification was based on growth patterns and colony morphology on selective media, alongside biochemical assays. To further validate phenotypic identification, representative presumptive S. aureus isolates were subjected to genotypic confirmation using polymerase chain reaction (PCR) (Chakravorty et al., 2007 ). Genomic DNA was extracted using a modified CTAB-based protocol, and PCR amplification targeting the rsmI gene was performed using species-specific primers under standard cycling conditions (Graber, 2016 ). Amplified products were resolved by agarose gel electrophoresis, and band sizes were compared with a molecular weight marker to confirm target amplification. Molecular confirmation was used to support phenotypic classification of S. aureus isolates recovered from smoked fish samples. Quantitative Microbial Risk Assessment Quantitative microbial risk assessment (QMRA) was determined using a four-step procedure to estimate the risk of response (foodborne illness or death) with respect to a known dose of a pathogen. The hazard identification, exposure assessment, dose-response, and risk characterization were evaluated using the Beta-Poisson model employing Firth's model alongside its correction bias. The QMRA was figuratively expressed from processing to consumption, and the probability of adverse health outcome was evaluated (Schelin et al., 2011 ; Pepper et al., 2015 ). Data Analysis All statistical analyses were conducted using Microsoft Excel 2019 and R statistical programming environment. Descriptive statistics were calculated for all variables. The normality of continuous variables was assessed using Shapiro-Wilk tests. One-way ANOVA followed by Tukey's Honest Significant Difference (HSD) post-hoc test was employed to compare mean bacterial counts across groups, as this test controls for family-wise error rate and is appropriate for multiple comparisons (Nanda et al., 2021 ; Kakati & Gogoi, 2024 ). For dose-response modeling, the beta-Poisson model was selected based on its established use for S. aureus risk assessment (Schelin et al., 2011 ) and its ability to model low-dose foodborne intoxication probabilities. The model parameters (α and β) were derived from selected literatures (Schelin et al., 2011 ; Sergelidis et al., 2014 ; Schelin et al., 2017 ; Pekana & Green, 2018 ). Firth’s penalized logistic regression was employed to reduce small‑sample bias and to handle sparse data and separation in the dose–response analysis, ensuring finite and more reliable coefficient estimates and confidence intervals (Heinze & Puhr, 2010 ; Rahman & Sultana, 2017 ; Walker & Smith, 2020 ). This approach reduces bias in maximum likelihood estimates when event rates are low. Sensitivity analysis varied α and β parameters by ± 20% of their estimated values to assess robustness of intoxication probability estimates. All analyses were conducted in R version 4.5.0 using packages: betareg, and logist. Statistical significance was set at p < 0.05 (Ogofure et al., 2024 ; Ogofure & Green, 2025 ; Ogofure et al., 2025 ). Results and Discussion Sociodemographic of Fish Smoking and Vendors in Benin City, Nigeria Unless otherwise stated, questionnaire-based analyses are reported for respondents with complete data ( n = 301), whereas microbiological and quantitative microbial risk assessment analyses were conducted on all smoked fish samples collected ( n = 330). The social demographic/socio-economic status of the participants or respondents (vendors) in the study is shown in Tables 1 a, b and c. Table 1 a. showed that about 44.19% of the respondents were aged 31–40 years, while 27.91% of the respondents were aged 21–30 years. All the respondents or fishmongers in the study location were females. About 65.12% of the respondents had primary education as their highest level of education, while 32.56% of the respondents had secondary as their highest level of education (Table 1 a). Only about 2.33% of the respondents had polytechnic education as their highest level of education. 79.07% of respondents were in full-time fish-selling businesses in the study location, while 18.6% were part-time fish sellers. Most of the fish sold in the study location were Scomber or Titus, which were sold by 39.87% of the respondents, while Meluza and Herring fish were sold by 33.22% and 18.27% of the respondents, respectively. 90.7% of the respondents participated in the smoking process of the fish sold in the study location, while 9.3% of the respondents did not participate in the smoking process. Similarly, 90.7% of the materials used for smoking the fish were of wood origin, while 4.65% were from wood shavings (sawdust) (Table 1 a). Table 1 a. Sociodemographic/Socioeconomic Status of the Participants ( n = 301) Variable Frequency Percentage Age 10-20yrs 7 2.33 21-30yrs 84 27.91 31-40yrs 133 44.19 41-50yrs 77 25.58 51-60yrs 0 0.00 Sex Male 0 0.00 Female 301 100.00 Level of Education Primary 196 65.12 Secondary 98 32.56 Polytechnic 7 2.33 Do You Sell Fish as Full Time, Part-Time, or Seasonal Hustle Full-time 238 79.07 Part-time 56 18.60 Seasonal hustle 7 2.33 Type of fish Sold Sese 16 5.32 Scomber/Titus 120 39.87 Meluza 100 33.22 Herring fish 55 18.27 Catfish 10 3.32 Directly Participation in the Smoking Process of the Fish Yes 273 90.70 No 28 9.30 Material used for smoking the fishes Wood smoke 273 90.70 Sawdust smoke 14 4.65 Others 14 4.65 Further socio-demographic characteristics is shown in Table 1 b. Most of the fish samples evaluated in the study were smoked on the same day (65.12%), while less than 30% (27.91%) were smoked a day before the analyses were carried out on the fish. Less than 5% and 3% of the fish were smoked 2 and 3 days before analyses, respectively. About 65.12% of the respondents revealed that they do not have any major preservation for the smoked fish, while 34.88% opined that they use a particular measure of preservation of the smoked fish. Most of the respondents were new to the fish business, as about 30.23% were between 1 and 2 years in the fish business, while about 35.58% had spent more than six years in the fish business (Table 1 b). Regardless, most respondents were unaware that the fish could be contaminated with germs and could serve as a reservoir of pathogens to man. This makes up about 65.12% of the respondents who claim to not know about the contamination of fish with germs. The materials used for wrapping the fish, as revealed by the respondents, were basically polythene materials, which 62.9% of the respondents had said they used for wrapping the smoked fish. After smoking the fish, on transit to the market, about 51.16% of the respondents opined that they do not cover the fish between the point of smoking and the point of sale. 48.84% of respondents reveal that they use a covering material for the fish (Table 1 b). Table 1 b. Sociodemographic/Socioeconomic Status of the Participants ( n = 301) Variable Frequency Percentage How long has the fish been smoked? Today 196 65.12 Yesterday 84 27.91 2 days ago 14 4.65 3 days ago 7 2.33 More than 3 days ago 0 0.00 Any measure of preservation for the smoked? Yes 105 34.88 No 196 65.12 Duration of Participants in the business of selling fish 1-2yrs 91 30.23 3-4yrs 84 27.91 5-6yrs 49 16.28 More than 6 years 77 25.58 Awareness or knowledge about the contamination of fish with germs Yes 105 34.88 No 196 65.12 Materials used for wrapping the smoked fish Paper 42 13.95 Cellophane 189 62.79 Both 70 23.26 Any covering for the fish after smoking in transit to the market? Yes 147 48.84 No 154 51.16 The socio-demographic/economic status of the participants is also shown in Table 1 c. The socio-demographics of the participants in the study, and concerning the materials used for covering the fish, (Table 1 c) revealed that 33.33% of the respondents sometimes wash these materials. In contrast, 28.75% of the respondents do not wash the materials at all, which could also be a source of contamination for the already smoked fish. Most of the fish sellers in the study usually smoke less than a hundred fish to be sold for the day, while 4.65% smoke about 200–300 fish to be sold for the day. The materials obtained from the materials used in the smoking of the fish, as reported by the respondents, were sourced locally by 18.06%. In comparison, 81% of the respondents said that they purchased the materials for smoking fish, such as cartons, cellophane, etc (Table 1 c). The rich tradition of smoking fish in Nigeria reflects not only its practical necessity for food preservation but also its cultural significance, which is deeply embedded in certain local communities. Table 1 c. Sociodemographic/Socioeconomic Status of the Participants ( n = 301) Variable Frequency Percentage Frequency of washing the covering material Very often 42 28.57 Sometimes 49 33.33 Once in a while 14 9.52 None at all 42 28.57 Very often 42 28.57 When last did Participants wash the covering material? Today 63 60.00 Yesterday 28 26.67 2 days ago 14 13.33 Number of fish smoked and Sold in a batch 300 fish 0 0.00 Where participants obtain the materials for fish processing Purchase them 245 81.40 Sources for them locally 56 18.60 Do Participants practice cleaning the fish surface with foam? Yes 266 88.37 No 35 11.63 How often do participants wash the foam to clean the fish? Everyday 91 34.21 Twice weekly 112 42.11 Once a month 49 18.42 Anytime 14 5.26 How often do participants make carton material for smoked fish? Once 133 44.19 Every time 126 41.86 Others 42 13.95 The smoking of fish, as an age-old practice, is passed down through generations, and it connects with societal structures and economic dynamics, shaping culinary traditions and livelihoods alike. Exploring the socio-demographic landscape of the smoked fish business exposes a plethora of interconnected factors, which are expressed in this study. Economic conditions play a pivotal role, as the trade of smoked fish often serves as a vital source of livelihood for many households, especially in coastal and riverine communities where fishing is a primary occupation (Adeyeye & Oyewole, 2016 ; Adeyeye, 2018 ). The socio-demographic and economic aspects of the smoked fish business in Nigeria are multifaceted and influenced by various factors, including cultural practices, economic conditions, and geographical location. Fish smoking is a skill that is learned over time, and older individuals in fishing communities may have extensive experience in traditional smoking methods (Omeje et al., 2022 ). The results of the socio-economic or demographics of fish mongers in this study were consistent with several reports in the literature about feminine domination of fish processing and smoking activities in many Nigerian communities, and most of these women were reported to have little or no formal education (Ayo-Olalusi et al., 2010 ; Omoruyi et al., 2016 ; Adetimehin et al. , 2019; Oduwale, 2019 ; Ayeloja et al., 2020 ; Ayeloja et al., 2021 ; Abolagba et al., 2023 ). While it is worthy of note that all fish vendors encountered in the study were females, Oduwale ( 2019 ) had the same statistics. However, at variance with the findings in this study, there are few reports about the involvement of men in the fish business or smoking activity (Ayeloja et al., 2021 ; Abolagba et al., 2023 ). While all studies agreed that the demographics of fish selling and smoking activity is largely tilted toward the feminine gender, Abolagba et al. ( 2023 ) opined that 70% of fish sellers were females while 30% were males. The report, however, varied with the findings in this study as all (100%) of the fish sellers were females. Concerning the education of fish vendors in the study, the results obtained were consistent with the report that about 65.5% (above half the population of fish vendors) had only primary education as their highest qualification. Similarly, findings by Oduwale ( 2019 ) revealed that over 50% of fish vendors in Benin City had Primary education as their highest level of education, while other studies, as in the reports of Adetimehin et al. (2019) and Ayeloja et al. ( 2021 ) revealed that more than 60% (combining no formal education and primary education) of fish vendors either had no formal education or primary education. Furthermore, the findings in this study concerning the age of fish vendors were consistent with the result obtained by Abolagba et al. ( 2023 ), who reported that 71% of farmers or sellers were within the age bracket of 25 to 40 years. While fish smoking is a primary occupation for some vendors, others may engage in it as a supplementary source of income (Adetimehin et al. , 2019; Abolagba et al., 2023 ). The aforementioned reports are consistent with the results obtained in this study, where some participants (fish vendors) claimed that the fish business is more like a side hustle for them, but others engage in the business as their primary source of income or livelihood. In this research, most of the vended smoked fish were Scomber, Hose mackerel and meluza, and this finding was consistent with the report by Oduwale ( 2019 ), who observed that Sese (Horse Mackerel), Scomber (Mackerel) and Meluza (Hake) were the predominant species of smoked fish sold in open markets in Edo State. Additionally, similar to the report mentioned earlier, Abolagba et al. ( 2023 ) opined that Scomber ( Scombar scombrus ) and Hake or Meluza ( Merluccius merluccius ) were amongst the preferred fish types smoked in Benin in addition to tilapia and catfish however, catfish ( Clarias gariepinus ) emerged as the preferred species of fish consumed in Benin City, Edo State. The above point by Abolagba et al. ( 2023 ) about the most preferred fish consumed in Benin City was at variance with the report of Oduwale ( 2019 ), who reported that Scomber and Hake were the most preferred fish sold in open markets in Edo State. Isolation of Coagulase-Positive and Negative Staphylococci from Smoked Fish The socio-demographic characteristics of fish vendors can significantly influence the microbiological quality of smoked fish with dire public health implications. Vendors with limited formal education may lack awareness of essential hygiene practices, increasing the risk of bacterial contamination during processing. Traditional smoking methods, often practiced by vendors, may not adhere to modern food safety standards, allowing pathogens like Staphylococcus aureus to thrive. The dominance of women in the fish-smoking trade, coupled with harsh economic realities and constraints, can further exacerbate contamination risk, as many vendors lack access to modern equipment or hygienic facilities. Additionally, because some vendors engage in fish smoking as a supplementary income source, they might employ some casual approach to hygiene and a plethora of inconsistent processing practices, which might also impact the microbiology and safety of the fish. The combination of these demographics can create an environment where harmful bacteria can proliferate, posing serious public health threats, especially in the form of foodborne illnesses. Therefore, addressing these socio-demographic challenges through better education, improved access to modern technologies, and targeted interventions is critical to safeguarding both food quality and public health. Based on the above premise, the coagulase-positive Staphylococcal counts in smoked fish samples, considering various socio-demographic and processing factors, revealed observable trends, although most differences were not statistically significant ( p > 0.05 ) (Supplementary Material S2.0 (Figure S1 )). Samples from fish vendors or respondents who were unaware that fish could act as a reservoir of bacterial pathogens exhibited higher Staphylococcal counts compared to those with knowledge of this fact. However, this difference was not statistically significant ( p > 0.05 ). The counts ranged from 2.56 ± 1.48 Log CFU/g in samples from vendors who were aware of the risk to 2.70 ± 1.34 Log CFU/g in samples from vendors without such knowledge [Supplementary Material S2.0 (Figure S1 )]. Similarly, the use of preservative measures was not statistically associated with Staphylococcal load in the smoked fish samples. Although our study found no statistically significant association between reported preservation practices and bacterial load (p > 0.05), this finding should not be interpreted as evidence that preservation is unimportant. The absence of statistical significance warrants cautious interpretation, as fewer than 35% of vendors reported using preservation measures, which likely limited statistical power to detect meaningful differences. Furthermore, the study design presents inherent limitations, including a small subgroup sample size, substantial heterogeneity in preservation methods ranging from simple covering to the use of chemical preservatives, and reliance on self-reported practices rather than direct observation, introducing potential measurement error. Vendor awareness was assessed through a single dichotomous question, which may not adequately capture the complexity of food safety knowledge, and residual confounding by unmeasured hygiene practices cannot be excluded. It is well-established that proper preservation and refrigeration are critical for preventing bacterial proliferation in smoked fish (Osibona & Amaechi, 2023 ; Ubadire-Agua et al., 2023 ; Huang et al., 2023 ). Our findings do not contradict this principle but rather highlight the need for more rigorous assessment of specific preservation methods and their implementation in real-world market settings. The bacterial counts ranged from 2.52 ± 1.42 Log CFU/g in fish where preservatives were used to 2.64 ± 1.46 Log CFU/g in fish without preservatives, indicating that no measurable reduction in bacterial load was observed within the context of this study [Supplementary Material S2.0 (Figure S2)]. When considering the type of smoking material used, no significant difference was observed between fish smoked using wood and those smoked using sawdust [Supplementary Material S2.0 (Figure S3)]. Both wood and sawdust originate from the same raw material, with sawdust being a byproduct of wood processing in mills. Consequently, their chemical composition, including key antimicrobial components such as phenols, carbonyls, and organic acids, is inherently similar, which likely accounts for the minimal variability observed in their microbial impact. However, both materials were included in our study to explore whether differences in particle size, combustion rate, and heat distribution during the smoking process could influence microbial contamination indirectly. These factors, while subtle, may have practical implications for food safety and quality. The Staphylococcal counts ranged from 2.02 ± 0.8 Log CFU/g for wood-smoked fish to 2.66 ± 1.3 Log CFU/g for sawdust-smoked fish, suggesting that the choice of smoking material had minimal impact on bacterial contamination [Supplementary material S2 (Fig. 3 ), p > 0.05 ]. In contrast, a significant difference in Staphylococcal load was noted based on the type of fish. Specifically, Meluza and Scomber fish exhibited significantly higher Staphylococcal counts compared to Catfish ( p < 0.05 ) [Supplementary Material S2.0 (Figure S4)]. However, no significant difference was found between other fish types, such as Herring and Mackerel. These findings underscore the role of fish type as a potential risk factor for Staphylococcus contamination in smoked fish products, highlighting the need for targeted interventions based on fish species to improve food safety. The duration between smoking and consumption is a critical yet often overlooked factor influencing the microbial quality of smoked fish, particularly for consumers who ingest the product without further cooking. This study evaluated the coagulase-positive Staphylococcal count of smoked fish samples based on the time elapsed between smoking and analysis, which serves as a proxy for the period before consumption. The Staphylococcal load varied significantly with the duration between smoking and analysis, ranging from 2.19 ± 1.48 Log CFU/g in fish analyzed on the same day of smoking to 3.59 ± 0.40 Log CFU/g in fish analyzed three days post-smoking [Supplementary Material S2.0 (Figure S5)]. Statistical analysis revealed a significant increase in Staphylococcal counts between fish evaluated on the same day of smoking and those evaluated after two and three days ( p < 0.05 ). Conversely, no significant difference was observed between fish analyzed on the same day of smoking and those analyzed the following day, suggesting a significant ( p < 0.05 ) bacterial growth surge occurs after the first 48 h. post-smoking. This finding highlights the rapid bacterial proliferation that can occur in smoked fish when left unrefrigerated or improperly stored for extended periods. The over one-log increase in bacterial counts observed after 48 h underscores the heightened risk of Staphylococcal contamination in fish that are consumed long after smoking. Such delays between smoking and consumption could pose serious public health risks, particularly in the context of coagulase-positive Staphylococcus , which is known to produce enterotoxins capable of causing foodborne illness. These results stress the importance of educating consumers on the risks associated with prolonged storage of smoked fish and emphasize the need for appropriate preservation. From a public health perspective, the higher microbial load in fish consumed days after smoking can elevate the risk of foodborne illnesses. The significant increase in bacterial count over 48 h suggests that smoked fish may become unsafe for consumption within just a few days if not properly stored. This is particularly concerning, especially for individuals consuming the fish without prior or proper processing. The ingestion of fish with high levels of coagulase-positive Staphylococcus species may lead to food poisoning (Martins et al., 2013 ). Therefore, consumers, especially those in informal markets where cold chain preservation may be limited, face heightened risks of foodborne illness if they consume smoked fish that has been improperly stored or left for several days post-smoking. Public health interventions should emphasize the importance of proper storage and timely consumption of smoked fish to minimize these risks. The coagulase-negative staphylococcal count of smoked fish samples based on the use of any preservative measure was found to range from 1.36 ± 1.20 Log CFU/g (fishes smoked without any measure of preservation) to 0.78 ± 1.04 (fishes smoked with some measure of preservation when not sold for the day) [Supplementary Material S2.0 (Figure S6)]. It was also revealed that there was a statistical difference ( p < 0.005 ) in the coagulase-negative staphylococcal count of smoked fish samples without some measure of preservation compared to fish samples from respondents who used some measure of preservation. The coagulase-negative staphylococcal count of smoked fish samples based on the awareness or knowledge of vendors about fish being a reservoir ranged from 1.77 ± 1.34 Log CFU/g (for vendors who know fish being a reservoir) to 1.96 ± 1.41 Log CFU/g (for vendors without knowledge of fish being a reservoir) [Supplementary Material S2.0 (Figure S7)]. It was observed that there was no statistical difference in the count of fish obtained from vendors with the awareness of smoked fish being a reservoir of bacteria. The coagulase-negative staphylococcal count based on the type of materials used for smoking (wood smoke and sawdust) ranged from 1.52 ± 1.11 Log CFU/g (fishes smoked using firewood) to 3.01 ± 0.58Log CFU/g (fishes smoked using sawdust) [Supplementary Material S2.0 (Figure S8)]. There was a statistical difference ( p < 0.05) in the count of smoked fish on the basis of the materials (wood and sawdust) used for smoking or processing of the fish. While coagulase-negative staphylococci are generally less pathogenic, their presence reflects hygiene and handling conditions and may contribute to spoilage and opportunistic risks. The coagulase-negative staphylococcal counts of smoked fish samples based on the fish types evaluated in the study ranged from 1.43 ± 1.10 Log CFU/g (Meluza) to 2.40 ± 1.51 Log CFU/g (Herring) [Supplementary Material S2.0 (Figure S9)]. Statistically, there was no significant difference ( p > 0.05 ) in the coagulase-negative staphylococcal counts of all the smoked fish samples evaluated in the study. The coagulase-negative staphylococcal counts of smoked fish samples on the basis of the duration of purchase before analysis were found to range from 1.61 ± 1.21 Log CFU/g (fishes smoked on the same day of analysis) to 2.16 ± 1.87 Log CFU/g (fishes smoked two days before analysis) [Supplementary Material S2.0 (Figure S10)]. It was observed that there was no statistical difference in the coagulase-negative staphylococcal counts of the smoked fish samples based on the duration of smoking before analysis. It has been reported that smoked fish, especially at the retail level, might be a potential source of foodborne bacterial pathogens if not properly handled (Ehizibolo et al., 2008 ; Abolagba & Igbinevbo, 2010 ). There are numerous reports in the literature concerning the contamination of smoked fish and ready-to-eat fish sold by fish vendors in local markets around different parts of Nigeria. The isolation of Staphylococcus from smoked fish (especially in the context of its burdens or estimates) is a critical aspect of food safety and public health, as Staphylococcus species are known pathogens with a reputation for causing foodborne illnesses (Ogofure & Igbinosa, 2021 ). At the same time, several studies have reported the presence of bacteria in smoked fish samples sold in open markets within Nigeria, only a limited number of studies evaluated and enumerated the burden or population of specific bacteria, such as Staphylococcus species in smoked fish or ready-to-eat fish. Akinwumi and Adegbehingbe ( 2015 ) evaluated the presence of S. aureus in smoked fish like Tilapia, Herring and Catfish. They opined that the staphylococcal count ranged from 1.4 × 10 4 Log CFU/g − 2.8 × 10 5 Log CFU/g and these counts were a product of poor handling practices and unsafe or unhygienic conditions in which the fish was smoked and exposed to the market. Edeh et al. ( 2023 ) also isolated Staphylococcus aureus from smoked fish from three different Nigerian markets in South-Eastern Nigeria, but they never quantified the staphylococcal burden of the evaluated samples; rather, their analyses were more of a qualitative evaluation. Jega et al. ( 2019 ) evaluated the microbiological burden of smoked fish sold at an open market in the Northern part of Nigeria and revealed that a certain sample had a Staphylococcal count of 5.4 × 10 5 Log CFU/g. They argued that a multitude of factors could be responsible for the poor microbiological quality of the smoked fish, which include the intrinsic environment where the fish was obtained before smoking, as well as the postharvest handling and processing methods employed by the sellers or vendors. Concerning the handling practices of fish vendors, Christoper et al. ( 2022 ) reported that the handling practices and hygienic conditions of smoked fish sold in open markets in Ado-Ekiti were critical to the presence of pathogenic Staphylococcus aureus in the smoked fish. It is worthy of note that the staphylococcal burden evaluated for both coagulase-positive and negative species in this study was found to be 2-log lesser than the aforementioned studies. Based on findings from this study, vendors are highly encouraged to imbibe simple handwashing practices with soap and clean water, make use of affordable and locally available disinfectants like vinegar or sanitizers for cleaning surfaces and equipment (Ogofure et al., 2018 ), and potentially adopt heat-based sterilization methods (e.g., boiling utensils). Additionally, the importance of basic hygiene training for vendors to increase awareness and encourage adherence to these practices cannot be overstated. Although the smoked fish samples in this study were not vacuum-packed, the observed low levels of S. aureus and coagulase-negative staphylococci (< 4 log CFU/g) align with previous findings in cold-smoked, vacuum-packed trout and salmon (Gonzalez-Rodriguez et al., 2002 ). However, caution should be exercised in interpreting these comparisons due to differences in packaging and storage conditions. The high salinity of the water in which the salmons were raised likely inhibited the growth of other competing bacteria, thereby creating conditions that favoured the proliferation of halophilic Staphylococcus species. However, the findings by Prasad et al. ( 2007 ) reported an average of 4.35–4.78 log Log CFU/g of total staphylococci in freshwater prawns, and this value was beyond values obtained in this study and also the permissible levels or limit for staphylococci in food. A few of the samples obtained in this study were found to be greater than a 2-log count (in terms of staphylococcal density) with respect to the permissible limits of Staphylococci in foods or smoked fish. Bergdoll (1979), as cited by Prasad et al. ( 2007 ), suggested that staphylococci count greater than 2-log Log CFU/g is a potential health hazard, and such food should be considered unsafe for consumption (although regulatory limits vary by countries and food products). This implies that going by the counts obtained in this study, most of the smoked fish samples, especially those smoked a day and 2 days before analyses, were above the permissible limit of log 2 Log CFU/g and thus unfit for consumption. Another study by Onjong et al. ( 2021 ) simulated the levels of S. aureus on slated, sun-dried, and smoked Nile tilapia fish. They revealed that the staphylococcal burden ranged from 4.0 log Log CFU/g to 9.01 log Log CFU/g. The presence of Staphylococcus species in smoked fish is expected because the bacteria can survive and grow in environments with low water activity and even in environments with high salt concentrations (Gonzalez-Rodriguez et al., 2002 ; Prasad et al., 2007 ). Similar to the findings in this study, where both coagulase-positive and coagulase-negative Staphylococcus isolates were detected in smoked fish, Hammad et al. ( 2012 ) reported comparable results from fish sold in grocery stores. Their study identified not only Staphylococcus aureus contamination but also pathogenic strains of coagulase-positive staphylococci, coagulase-negative staphylococci, methicillin-resistant staphylococci, and methicillin-resistant coagulase-negative staphylococci. These observations underscore the widespread prevalence of both coagulase-positive and coagulase-negative staphylococcal species in various fish products, highlighting significant food safety concerns and the potential for antibiotic-resistant strains to pose additional public health risks. Most of the reports made use of mannitol salt agar, which has long been employed for isolating staphylococci, as described by Bridson ( 2006 ). Previous studies have expanded traditional mannitol salt agar workflows by incorporating additional selective media for isolation of presumptive pathogenic staphylococci (Ogofure & Igbinosa, 2021 ; Mues et al., 2025 ). They included Vogel Johnson agar (which differentiates coagulase-positive Staphylococcus aureus ) and oxacillin-resistant screening agar to identify methicillin-resistant staphylococci. This multi-faceted approach enhanced the precision of isolating and characterizing staphylococcal species, particularly in differentiating antibiotic-resistant strains, offering a more comprehensive tool for staphylococcal detection in food microbiology contexts. In this study, we decided to increase the salinity of the saline solution to 7.5% w/v to achieve more precise isolation of halophilic Staphylococcus species while employing the sequence of the three-differential media. This elevated salinity level was intended to selectively promote the growth of halotolerant staphylococci while discouraging the proliferation of other non-halophilic microbial isolates. This modification further refined the isolation process, ensuring a more targeted and accurate assessment of staphylococcal contamination in smoked fish. The prevalence of various Staphylococcal isolates from smoked fish is shown in Fig. 1 . Remarkably, 97.78% of the smoked fish samples were found to be contaminated with Staphylococcus species. Among these, 46.67% were contaminated with coagulase-positive Staphylococcus aureus , while 64.44% harboured coagulase-negative Staphylococcus species. Furthermore, 37.78% of the samples were contaminated with methicillin-resistant Staphylococcus aureus (MRSA) (Fig. 1 .). These findings underscore the widespread contamination of smoked fish with both pathogenic and antimicrobial-resistant Staphylococcal species, highlighting potential public health risks associated with the consumption of improperly handled or processed fish. The prevalence of Staphylococcus species has been widely reported in the literature, with significant variations across different studies. Hammad et al. ( 2012 ) observed a notably comparable higher prevalence of Staphylococcus species (87%), including pathogenic S. aureus and methicillin-resistant Staphylococcus aureus (MRSA), in smoked fish samples. These findings differ considerably from the results in our study, where the prevalence of coagulase-positive S. aureus and MRSA was recorded at 46.67% and 37.78%, respectively. Similarly, Grigoryan et al. ( 2010 ) reported a higher prevalence of S. aureus (74%) during cold-smoked fish processing, identifying the brining stage as a critical control point for preventing contamination. In contrast to the findings in our study, Vazquez-Sanchez et al. ( 2012 ) found a lower prevalence of S. aureus in smoked fish samples at 26%, with higher contamination levels observed in fresh and frozen products. Similar to the aforementioned study, Pekana and Green ( 2018 ) observed a lower prevalence rate (less than 21%) of Staphylococcus species from frozen meat products in South Africa. This may suggest that improved regulatory oversight could mitigate similar contamination risks in Benin City. Furthermore, a study by Karimela et al. ( 2017 ) highlighted that S. aureus was a dominant contaminant in traditionally smoked scad fish from Sangihe Island, with a prevalence of 62%. These comparative findings emphasize the variability in Staphylococcus contamination across different smoking practices and processing stages, underscoring the need for stringent control measures to reduce the risk of contamination in smoked fish products. Microbial Risk Assessment of Smoked Fish Contaminated with Staphylococcus species The quantitative microbial risk assessment (QMRA) of Staphylococcus species in smoked fish, providing a critical evaluation of the risk posed by Staphylococcal contamination to consumers, is shown in Fig. 2. QMRA typically follows a four-step framework, hazard identification, hazard characterization, exposure assessment, and risk characterization, each of which was employed in this study to estimate the probability of adverse health outcomes following the consumption of smoked fish contaminated with Staphylococcus species. In our assessment, the ß-Poisson model was used to characterize the dose-response relationship, which quantifies the probability of adverse health outcome based on the level of bacterial exposure. Our study found a prevalence of Staphylococcal isolates in 97.8% of all smoked fish samples, with 46.67% of these isolates identified as Staphylococcus aureus post-smoking (Fig. 2). The beta-Poisson model describes the probability of adverse health outcome ( P ) as a function of the dose ( D ) of a pathogen. The model equation is given by: \(\:\:P=1-{e}^{-(\beta\:.D)\alpha\:}\) Figure 2. Graphical representation of quantitative microbial risk assessment of staphylococcal isolates from smoked fish Most smoked fish are usually purchased within three days of smoking and are sometimes consumed without further processing, significantly increasing the risk of bacterial transmission, as reported by our earlier findings. The probability of adverse health outcome relative to doses of staphylococcal isolates consumed from smoked fish is shown in Fig. 3 . Utilizing Firth's model to predict the probability of adverse health outcome, it was observed that there is a high likelihood of infection, exceeding 90%, when consumers are exposed to higher doses of Staphylococcus aureus in smoked fish (Fig. 3 ). This risk is particularly pronounced for individuals consuming fish that has been smoked and left for 2–4 days before being sold (particularly those purchased greater than 2 days post-smoking). The model suggests that the risk of infection remains alarmingly high, even exceeding 80%, under typical storage and consumption conditions. These findings emphasize the critical need for improved handling, storage, and consumer awareness to mitigate the risk of staphylococcal infection. Consumer awareness can be enhanced through community outreach programs, media campaigns utilizing accessible platforms like radio and social media, and collaborations with vendors to educate consumers about proper storage practices, the dangers of consuming contaminated smoked fish, and how to identify spoiled products. The prolonged shelf life of smoked fish, particularly under non-refrigerated conditions, significantly increases the probability of bacterial growth and the subsequent risk of foodborne illness. In essence, while smoking helps preserve fish, it doesn't make it completely safe for extended periods, especially without proper refrigeration. The longer smoked fish is kept, particularly at room temperature, the higher the risk becomes of it harbouring harmful bacteria, in this case, Staphylococcus species. This underscores the necessity of immediate consumption or proper refrigeration practices to reduce the public health threat posed by Staphylococcus aureus in smoked fish product. The possibility of staphylococcal contamination of smoked fish is a serious threat to public health even though Sikorski and Kalodziejska ( 2002 ) asserted that contamination of smoked seafood by Staphylococcus aureus is rare. Recent studies have increasingly highlighted the issue of Staphylococcus contamination in food products, including smoked fish. Several researches such as Christoper et al. ( 2022 ), Edeh et al. ( 2023 ), and Musa et al. ( 2020 ) reported that smoked fish can indeed become contaminated, sometimes with astronomical bacterial burdens. These findings underscore the need for heightened vigilance in food safety practices, particularly regarding smoked fish, as the risk of contamination appears to be rising with an increasing number of days post-smoking. Despite these possibilities of Staphylococcal contamination, poor processing and handling of raw and smoked fish can lead to contamination by Staphylococcus species, which in turn can favour toxin production, increasing the risk of infection in consumers. The vulnerability of smoked fish to Staphylococcus contamination has been widely reported, highlighting the necessity for a comprehensive risk assessment. Our study demonstrated that the risk characterization and probability of illness from the consumption of smoked fish contaminated with Staphylococcus species is dose-dependent. A significant correlation was found between the Staphylococcus load and various factors, particularly the time elapsed between smoking and consumption. Fish smoked two days post-purchase were observed to harbour a higher microbial load compared to freshly smoked samples, thereby increasing the dose per serving and the subsequent risk of infection. This observation aligns with the findings of Onjong et al. ( 2021 ), who reported an elevated risk of staphylococcal enterotoxin production in smoked Nile tilapia, particularly in unregulated value chains in Kenya. Their study noted that the time taken to sell fish, whether under ambient or refrigerated conditions, directly influenced the dose-per-serving and enterotoxin production. This parallels our findings, where Staphylococcal load increased with duration post-smoking from 24 to 72 h, and the time spent selling the product contributed further to the risk of toxin production. Additionally, Musa et al. ( 2020 ) identified Staphylococcus aureus contamination in smoked fish sold at Ahmadu Bello University, underscoring the health risks posed to consumers. Sergelidis et al. ( 2014 ) reported that 43% of ready-to-eat fish products, including smoked varieties, present a potential health hazard due to the presence of methicillin-resistant Staphylococcus aureus (MRSA). To mitigate the risk of Staphylococcus contamination and toxin production, strict adherence to hygiene and best practices during both the smoking and vending processes is critical. Vendors should employ better preservation techniques for unsold fish to prevent microbial proliferation. Furthermore, consumer awareness programs are essential to prevent the possibility of Staphylococcal food poisoning outbreaks. Study Limitations This study has some limitations that should be considered when interpreting the findings. Although 330 smoked fish samples were analyzed microbiologically, questionnaire-based analyses were limited to respondents with complete data, which may have reduced statistical power for some comparisons. In addition, information on handling and preservation practices was self-reported and may be subject to recall or social desirability bias. The cross-sectional design also limits causal inference between vendor practices and microbial contamination levels. Nevertheless, the combined use of microbiological analyses and quantitative microbial risk assessment provides a robust, context-specific evaluation of public health risks associated with smoked fish sold in informal markets. Conclusion and Recommendation This study provides the first comprehensive quantitative microbial risk assessment of S. aureus in artisanally smoked fish from randomly selected markets in Edo South senatorial district, revealing alarmingly high MRSA prevalence (37.78%) and demonstrating critical time-dependent contamination thresholds. While bacterial proliferation in unrefrigerated food is well-established, our study uniquely quantifies infection probability at specific consumption timepoints (> 90% risk at 72 h post-smoking) and identifies fish species-specific vulnerability patterns not previously documented in this context. These estimates are model-based and reflect worst-case exposure scenarios under ambient storage conditions. These results reinforce the urgent need for public health efforts, which focus on introducing low-cost refrigeration or insulated storage solutions for vendors, alongside training on hygienic handling and storage methods and increased consumer awareness to minimize the risks associated with smoked fish. More so, immediate consumption or proper refrigeration should be emphasized to reduce the potential for bacterial contamination, particularly with coagulase-positive and methicillin-resistant strains. Our study contributes significantly to UN Sustainable Development Goals (SDGs) 2 (Zero Hunger), 3 (Good Health and Well-being), and 12 (Responsible Consumption and Production). By assessing microbial risks and processing practices of smoked fish, our study directly addresses food safety concerns (SDG 2.1: ensuring access to safe and nutritious food) and promotes public health (SDG 3.3: combating communicable diseases). The identification of critical factors influencing staphylococcal contamination in smoked fish supports the development of sustainable food production systems (SDG 2.4) and encourages responsible consumption patterns (SDG 12.3: reducing food losses along production and supply chains). Our findings on the high prevalence of antibiotic-resistant strains (MRSA) contribute to the global effort to combat antimicrobial resistance (SDG 3.d). By emphasizing the need for enhanced consumer education, improved preservation practices, and stricter regulations, this research supports the implementation of sustainable consumption and production patterns (SDG 12.8: ensuring awareness for sustainable development). Ultimately, these scientific observations provide crucial insights for policymakers, food producers, and consumers to make informed decisions that enhance food safety, reduce health risks, and promote sustainable practices in the fish-smoking industry. Abbreviations QMRA Quantitative Microbial Risk Assessment MRSA methicillin-resistant Staphylococcus aureus SDGs Sustainable Development Goals SFP Staphylococcal food poisoning WHO World Health Organization LGA Local Government Areas HSD Honest Significant Difference CoNS Coagulase Negative Staphylococci Declarations Conflict of Interest The authors have no relevant financial or non-financial interests to disclose. Funding Statement This research was funded by the University of Johannesburg University Research Committee, Project No. 075432. Authors’ contributions Progress E. Boyo-Agbonile, Abraham G. Ogofure, Brenda O. Isichel-Ukah, Ezekiel Green and Etinosa O. Igbinosa contributed to the study conception and design. Progress E. Boyo-Agbonile, Abraham G. Ogofure, Brenda O. Isichel-Ukah and Etinosa O. Igbinosa performed material preparation, data collection and analysis. Progress E. Boyo-Agbonile, Abraham G. Ogofure, and Etinosa O. Igbinosa, wrote the first draft of the manuscript, and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript. Acknowledgements The authors are grateful for the material support from the Department of Biotechnology and Food Technology, University of Johannesburg, South Africa, toward the success of this research. We thank the Alexander von Humboldt Foundation for the equipment grant awarded to Prof. E.O. Igbinosa. More importantly, authors are grateful to God for directing our steps in this research, knowing that “it is God who works in us to will and to act” (Philippians 2:13). Data Availability Statement The data underlying this manuscript are available in the manuscript and in its supplementary material. References Abdi AM, Amano A, Abrahim A, Getahun M, Ababor S, Kumie A (2020) Food Hygiene Practices and Associated Factors Among Food Handlers Working in Food Establishments in the Bole Sub City, Addis Ababa, Ethiopia. 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Fish Technol 44:22–29. https://doi.org/http://krishi.icar.gov.in/jspui/handle/123456789/56336 Puah SM, Chua KH, Tan JA (2016) Virulence Factors and Antibiotic Susceptibility of Staphylococcus aureus Isolates in Ready-to-Eat Foods: Detection of S. aureus Contamination and a High Prevalence of Virulence Genes. Int J Environ Res Public Health 13(2):199. https://doi.org/10.3390/ijerph13020199 Rahman MS, Sultana M (2017) Performance of Firth-and logF-type penalized methods in risk prediction for small or sparse binary data. BMC Med Res Methodol 17(1):33. https://doi.org/10.1186/s12874-017-0313-9 Schelin J, Susilo YB, Johler S (2017) Expression of Staphylococcal Enterotoxins under Stress Encountered during Food Production and Preservation. Toxins (Basel) 9(12). https://doi.org/10.3390/toxins9120401 Schelin J, Wallin-Carlquist N, Cohn MT, Lindqvist R, Barker GC, Radstrom P (2011) The formation of Staphylococcus aureus enterotoxin in food environments and advances in risk assessment. Virulence 2(6):580–592. https://doi.org/10.4161/viru.2.6.18122 Sergelidis D, Abrahim A, Papadopoulos T, Soultos N, Martziou E, Koulourida V, Govaris A, Pexara A, Zdragas A, Papa A (2014) Isolation of methicillin-resistant Staphylococcus spp. from ready-to-eat fish products. Lett Appl Microbiol 59(5):500–506. https://doi.org/10.1111/lam.12304 Shajil S, Mary A, Rani-Juneius CE (2018) Recent Food Preservation Techniques Employed in the Food Industry. In Microbial Biotechnology (pp. 3–21). https://doi.org/10.1007/978-981-10-7140-9_1 Sikorski ZE, Kalodziejska I (2002) Microbial risks in mild hot smoking of fish. Crit Rev Food Sci Nutr 42(1):35–51. https://doi.org/10.1080/10408690290825448 Teffo LA, Tabit FT (2020) An assessment of the food safety knowledge and attitudes of food handlers in hospitals. BMC Public Health 20(1):311. https://doi.org/10.1186/s12889-020-8430-5 Ubadire-Agua C, Ezeama CF, Anyiam CA, Anyiam CC, Nwanagba NL (2023) Effect of Some Preservatives on the Microbial Quality of Cold-smoked Catfish at Refrigerated Temperature. Asian Food Sci J 22(10):1–11. https://doi.org/10.9734/afsj/2023/v22i10668 Vazquez-Sanchez D, Lopez-Cabo M, Saa-Ibusquiza P, Rodriguez-Herrera JJ (2012) Incidence and characterization of Staphylococcus aureus in fishery products marketed in Galicia (Northwest Spain). Int J Food Microbiol 157(2):286–296. https://doi.org/10.1016/j.ijfoodmicro.2012.05.021 Walker DA, Smith TJ (2020) Logistic Regression Under Sparse Data Conditions. J Mod Appl Stat Methods 18(2):2–18. https://doi.org/10.22237/jmasm/1604190660 Wu S, Duan N, Gu H, Hao L, Ye H, Gong W, Wang Z (2016) A Review of the Methods for Detection of Staphylococcus aureus Enterotoxins. Toxins (Basel) 8(7). https://doi.org/10.3390/toxins8070176 Supplementary Files BoyoAgbonileetalSupplementaryMaterials2025.30.12.docx Cite Share Download PDF Status: Published Journal Publication published 01 May, 2026 Read the published version in Annals of Microbiology → Version 1 posted Reviewers agreed at journal 20 Jan, 2026 Reviewers invited by journal 20 Jan, 2026 Editor assigned by journal 05 Jan, 2026 First submitted to journal 02 Jan, 2026 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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20:25:10","extension":"html","order_by":14,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":229408,"visible":true,"origin":"","legend":"","description":"","filename":"earlyproof.html","url":"https://assets-eu.researchsquare.com/files/rs-8482256/v1/b71880e9d2cccbe7eadc4019.html"},{"id":100921980,"identity":"e873c19a-c6b8-4c22-98ab-a7da231b2820","added_by":"auto","created_at":"2026-01-22 20:25:11","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":87792,"visible":true,"origin":"","legend":"\u003cp\u003ePrevalence of the different staphylococcal isolates from smoked fish samples\u003c/p\u003e","description":"","filename":"floatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-8482256/v1/0f1f096e193539d4030c09fe.png"},{"id":100921982,"identity":"a2d76f49-6fad-49ce-8049-864e8ce51df2","added_by":"auto","created_at":"2026-01-22 20:25:11","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":726148,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eGraphical representation of quantitative microbial risk assessment of staphylococcal isolates from smoked fish\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"floatimage2.png","url":"https://assets-eu.researchsquare.com/files/rs-8482256/v1/d25fce28b2eede4182d425da.png"},{"id":100921970,"identity":"ba74d4c4-3585-42a5-bb16-7de6daf5d23f","added_by":"auto","created_at":"2026-01-22 20:25:10","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":50824,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eProbability of adverse health outcome relative to doses of staphylococcal isolates consumed from smoked fish\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"floatimage3.png","url":"https://assets-eu.researchsquare.com/files/rs-8482256/v1/6a9e9f8461a25674501f053e.png"},{"id":108437956,"identity":"df3b7fd0-9d24-4495-8667-013651a68dbe","added_by":"auto","created_at":"2026-05-04 16:04:48","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1453682,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8482256/v1/88da467d-f952-45a2-814c-6ec431dffa7d.pdf"},{"id":100921976,"identity":"b00900af-5456-41a7-b02e-eed848cc36de","added_by":"auto","created_at":"2026-01-22 20:25:10","extension":"docx","order_by":5,"title":"","display":"","copyAsset":false,"role":"supplement","size":572704,"visible":true,"origin":"","legend":"","description":"","filename":"BoyoAgbonileetalSupplementaryMaterials2025.30.12.docx","url":"https://assets-eu.researchsquare.com/files/rs-8482256/v1/ea33f6c7e29ec1d2cac0907e.docx"}],"financialInterests":"","formattedTitle":"Microbial risk assessment of Staphylococcus species in smoked fish from informal markets in Edo State, Nigeria: processing practices and public health implications","fulltext":[{"header":"Introduction","content":"\u003cp\u003eFish is a critical source of high-quality protein, micronutrients, and essential fatty acids, making it a dietary staple for millions of people globally (Obiero et al., \u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Mohanty et al., \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Aremu et al., \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Ekonomou et al., \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2022\u003c/span\u003e), particularly in regions like Nigeria. However, postharvest losses due to microbial spoilage are a significant concern, especially in developing countries where traditional fish preservation methods dominate (Ghaly, \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Diop et al., \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Aremu et al., \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Smoking is one of the most common preservation techniques in these regions, utilized not only to enhance flavour and texture but also to prolong shelf life by reducing moisture content and inhibiting microbial growth (Shajil et al., \u003cspan citationid=\"CR66\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Olusola, \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Ekonomou et al., \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Deliephan et al., \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). While this method has proven to be effective against some spoilage organisms, it may also create an environment conducive to contamination by pathogens such as \u003cem\u003eStaphylococcus aureus\u003c/em\u003e, posing a substantial risk to public health (Ghaned et al., \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Musa et al., \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Doutoum et al., \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cem\u003eStaphylococcus aureus\u003c/em\u003e is a facultative anaerobic bacterium and opportunistic pathogen that thrives in various environmental conditions and is commonly associated with foodborne illnesses (Ogofure \u0026amp; Igbinosa, \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Merghni et al., \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). The bacterium produces a wide range of virulence factors, including heat-stable enterotoxins, which can persist in smoked fish even after processing (Puah et al., \u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Fisher et al., \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). In developing countries, the challenges of ensuring hygienic food handling practices are exacerbated by limited regulatory oversight, inadequate infrastructure, and the widespread use of artisanal processing methods (Alimi, \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Morse et al., \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Abdi et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). As such, smoked fish sold in open markets across Nigeria may become a reservoir for pathogenic bacteria, including coagulase-positive \u003cem\u003eStaphylococcus\u003c/em\u003e species like \u003cem\u003eS. aureus\u003c/em\u003e, whose heat-stable toxin can remain viable under cooking or cells can survive suboptimal cooking or processing temperatures.\u003c/p\u003e \u003cp\u003eConsumption of smoked fish contaminated with \u003cem\u003eS. aureus\u003c/em\u003e can pose a significant threat to public health, as the ingestion of staphylococcal enterotoxins can lead to staphylococcal food poisoning (SFP), characterized by nausea, vomiting, abdominal cramps, and diarrhea (Wu et al., \u003cspan citationid=\"CR72\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Schelin et al., \u003cspan citationid=\"CR63\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Guo et al., \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Given the widespread reliance on smoked fish as a protein source in Nigeria, understanding the extent of microbial contamination and the associated risk to consumers is imperative. In addition to enterotoxin production, antibiotic-resistant strains of \u003cem\u003eS. aureus\u003c/em\u003e, particularly methicillin-resistant \u003cem\u003eS. aureus\u003c/em\u003e (MRSA), are becoming an increasing concern globally (Liu et al., \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Nandhini et al., \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). These strains complicate treatment options for foodborne illness and contribute to the growing burden of antimicrobial resistance (AMR), a public health crisis recognized by the World Health Organization (WHO) (Liu et al., \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2021\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eIn Benin City, Nigeria, limited access to refrigeration and reliance on artisanal preservation practices heighten the risk of bacterial contamination in smoked fish, a dietary staple. Thus, while smoked fish is widely consumed, there is limited information on the prevalence of \u003cem\u003eS. aureus\u003c/em\u003e in our study location. Furthermore, the identification of the specific processing practices that may contribute to bacterial contamination, and evaluation of the probability of adverse health outcome based on consumption quantity or patterns will be crucial to assess the probable risk of \u003cem\u003eS. aureus\u003c/em\u003e in smoke fish staple. This study addresses a critical gap in the literature by conducting a comprehensive microbial risk assessment of \u003cem\u003eS. aureus\u003c/em\u003e-contaminated smoked fish sold in Benin City, Nigeria, and the hygienic conditions (processing practices) from the vendor's perspective that may impact consumer health.\u003c/p\u003e \u003cp\u003eTherefore, we aim to provide a detailed assessment of fish processing practices and the associated microbial risks with emphasis on \u003cem\u003eStaphylococcus\u003c/em\u003e species contamination from fish samples, specifically looking at Coagulase positive \u003cem\u003eStaphylococcus aureus\u003c/em\u003e, methicillin resistant \u003cem\u003eStaphylococcus aureus\u003c/em\u003e and coagulase negative staphylococci. Additionally, our study examines the public health implications, particularly in the context of food safety regulations. By integrating microbial risk assessment models with an analysis of vendor practices, this work aims to inform policy development and promote safer food processing methods, ultimately reducing the burden of foodborne illness and supporting sustainable food systems in Nigeria and beyond.\u003c/p\u003e"},{"header":"Methodology","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eStudy Design and Location\u003c/h2\u003e \u003cp\u003eThis study was conducted across twenty (20) open markets located in three Local Government Areas (LGA) within the Edo South Senatorial district of Benin City, Nigeria. Smoked fish samples were collected from these markets and categorized according to local terminology into different groups, which include Scomber or Titus fish (\u003cem\u003eScomber scombrus\u003c/em\u003e), Hake or Meluza (\u003cem\u003eMerluccius merluccius\u003c/em\u003e), Catfish (\u003cem\u003eClarias gariepinus\u003c/em\u003e), Sese or Horse Mackerel (\u003cem\u003eTrachurus trachurus\u003c/em\u003e) and Herring or Shawa (\u003cem\u003eClupea harengus\u003c/em\u003e). Heterotrophic bacterial counts were determined, and staphylococci isolates were differentiated into coagulase-negative, coagulase-positive, methicillin-resistant \u003cem\u003eStaphylococcus aureus\u003c/em\u003e (MRSA), and non-MRSA groups. The prevalence of pathogenic staphylococcal isolates was also evaluated. In addition, a structured questionnaire was administered to vendors to assess postharvest handling practices and hygiene-related behaviors associated with smoked fish processing. The questionnaire is provided in the Supplementary Material (S1.0). All questionnaire responses were anonymized, and no personally identifiable information was collected.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eSampling Strategy\u003c/h3\u003e\n\u003cp\u003eIn this study, a multi-stage sampling approach was employed covering three Local Government Areas (LGAs) in the Edo South Senatorial district, which were purposively selected based on high fish trading activity and large population density. Within each LGA, open markets were randomly selected from an official list of markets, yielding a total of twenty markets. Within each market, vendors were approached at random until the target sample size was achieved. A total of 405 vendors were approached, of whom 330 agreed to participate and provided written informed consent, resulting in a response rate of 81.48%. Vendors who declined participation cited time constraints or privacy concerns and were excluded from the study. Each participating vendor provided one smoked fish sample, resulting in a total of 330 fish samples collected for microbiological analysis.\u003c/p\u003e\n\u003ch3\u003eQuestionnaire Design, Administration, and Data Processing\u003c/h3\u003e\n\u003cp\u003eA structured questionnaire was designed and developed based on existing literature on food safety practices (Teffo \u0026amp; Tabit, \u003cspan citationid=\"CR68\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Halabi et al., \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Alqadiri et al., \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2025\u003c/span\u003e) and refined through expert consultation with food microbiologists and public health specialists in the Microbiology Postgraduate Board, University of Benin (during proposal presentation). The questionnaire was designed to assess postharvest handling practices, hygienic behaviors and conditions, and vendor awareness regarding bacterial contamination of smoked fish. The questionnaire comprised four key construct domains which include (1) demographic variables, to evaluate socioeconomic factors influencing hygiene practices; (2) processing practices, to assess potential risk factors associated with product handling; (3) preservation and storage, to identify critical control points related to product safety; and lastly (4) hygiene awareness, a recognized predictor of food safety behaviors (Abdi et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Teffo \u0026amp; Tabit, \u003cspan citationid=\"CR68\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Alqadiri et al., \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2025\u003c/span\u003e). Prior to data collection, the questionnaire was pre-tested with five smoked fish vendors from a market outside the study area to assess clarity, relevance, and contextual appropriateness. Feedback from the pilot testing informed minor revisions before final deployment. The finalized questionnaire (Supplementary Material S1.0) was administered in person by trained research assistants fluent in English and the local dialect (Bini), enabling participation by vendors with limited literacy. Questionnaires were administered at vendors\u0026rsquo; points of sale after participants had received an explanation of the study objectives and procedures and had provided written informed consent.\u003c/p\u003e \u003cp\u003eA total of 330 vendors participated in the survey, corresponding to the number of smoked fish samples collected. However, some questionnaires contained incomplete or internally inconsistent responses for one or more variables. To ensure analytical validity, only questionnaires with complete responses for the relevant variables were included in the final descriptive and inferential analyses (complete-case analysis), resulting in an effective questionnaire sample size of 301 respondents. All microbiological analyses were conducted on the full set of 330 smoked fish samples. Completed questionnaires were anonymized at the point of collection, coded, and entered into Microsoft Excel for data cleaning prior to statistical analysis\u003c/p\u003e\n\u003ch3\u003eSample Collection\u003c/h3\u003e\n\u003cp\u003eUsing a prevalence rate of 26%, as observed in similar studies from Nigeria (Vazquez-Sanchez et al., \u003cspan citationid=\"CR70\" class=\"CitationRef\"\u003e2012\u003c/span\u003e), the sample size was determined to ensure adequate statistical power. The sample size and parameters were determined using the formula.\u003cdiv id=\"Equa\" class=\"Equation\"\u003e\u003cdiv format=\"TEX\" class=\"mathdisplay\" id=\"FileID_Equa\" name=\"EquationSource\"\u003e\n$$\\:Sample\\:size\\:\\left(N\\right)=\\:{(Z}^{2\\:}P\\:\\left(1-P\\right))\u0026divide;{D}^{2}\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:\\:(1)$$\u003c/div\u003e\u003c/div\u003e\u003c/p\u003e \u003cp\u003ewhere: \u003cem\u003eN\u0026thinsp;=\u0026thinsp;Sample size, Z\u0026thinsp;=\u0026thinsp;z-score (1.969), P\u0026thinsp;=\u0026thinsp;estimated prevalence (0.26) and D\u0026thinsp;=\u0026thinsp;0.05\u003c/em\u003e (margin of error). Based on this calculation, a minimum sample size of 299 was obtained. This value was increased to 330 to account for potential non-response and incomplete data. The weight of each fish sample was recorded, and visual inspection of the smoked fish was conducted prior to microbiological analysis.\u003c/p\u003e\n\u003ch3\u003eStaphylococcal Culture and Enumeration\u003c/h3\u003e\n\u003cp\u003eTwenty-five grams (25 g) of each smoked fish sample were aseptically weighed and homogenized separately in two different sterile diluents to support differential microbial enumeration. For total heterotrophic bacterial counts, samples were homogenized in 225 mL of sterile physiological saline (0.85% w/v NaCl). In parallel, a separate homogenate of 25 g of the same sample was prepared in 225 mL of sterile high-salt saline (7.5% w/v NaCl) to favour the recovery of halotolerant staphylococci while suppressing the growth of non-halotolerant background microbiota. Each homogenate was thoroughly mixed, and tenfold serial dilutions were prepared using the corresponding saline diluent. From the 0.85% saline series, 1.0 mL aliquots from appropriate dilutions (typically 10\u003csup\u003e⁻\u0026sup3;\u003c/sup\u003e) were plated onto nutrient agar using the pour-plate method for total heterotrophic bacterial enumeration. From the 7.5% saline series, 1.0 mL aliquots from lower dilutions (typically 10\u003csup\u003e⁻1\u003c/sup\u003e) were plated onto Mannitol Salt Agar (MSA) for the selective enumeration of presumptive staphylococci. All inoculated plates were incubated aerobically at 37\u0026deg;C for 24 h. Following incubation, colonies were counted on plates containing 30\u0026ndash;300 colonies and results were expressed as colony-forming units per gram (CFU/g) of sample. On MSA, presumptive staphylococci were identified based on their ability to grow under high-salt conditions and by mannitol fermentation reactions, indicated by yellow (mannitol-positive) or pink (mannitol-negative) colonies. Representative colonies exhibiting distinct morphologies were subcultured onto fresh nutrient agar to obtain pure isolates for subsequent phenotypic (Bridson, \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2006\u003c/span\u003e; Ogofure \u0026amp; Igbinosa, \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). All experiments were conducted in three independent biological replicates. Based on the lowest dilution plated and a plated volume of 1.0 mL, the detection limit of the enumeration procedure was 10 CFU/g (1.0 log₁₀ CFU/g).\u003c/p\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eCharacterization of Staphylococcal Isolates from Smoked Fish\u003c/h2\u003e \u003cp\u003ePresumptive staphylococcal isolates initially enumerated on Mannitol Salt Agar were further characterized phenotypically. Coagulase-positive staphylococci were identified using Vogel-Johnson Agar (Oxoid), supplemented with 1% potassium tellurite solution, which allows for differentiation based on tellurite reduction and mannitol fermentation. For the detection of methicillin-resistant \u003cem\u003eStaphylococcus aureus\u003c/em\u003e (MRSA), isolates were cultured on Oxacillin Resistance Screening Agar containing oxacillin supplement, following standard guidelines (Bridson, \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2006\u003c/span\u003e; Ogofure \u0026amp; Igbinosa, \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Phenotypic identification was based on growth patterns and colony morphology on selective media, alongside biochemical assays. To further validate phenotypic identification, representative presumptive \u003cem\u003eS. aureus\u003c/em\u003e isolates were subjected to genotypic confirmation using polymerase chain reaction (PCR) (Chakravorty et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2007\u003c/span\u003e). Genomic DNA was extracted using a modified CTAB-based protocol, and PCR amplification targeting the \u003cem\u003ersmI\u003c/em\u003e gene was performed using species-specific primers under standard cycling conditions (Graber, \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Amplified products were resolved by agarose gel electrophoresis, and band sizes were compared with a molecular weight marker to confirm target amplification. Molecular confirmation was used to support phenotypic classification of \u003cem\u003eS. aureus\u003c/em\u003e isolates recovered from smoked fish samples.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eQuantitative Microbial Risk Assessment\u003c/h3\u003e\n\u003cp\u003eQuantitative microbial risk assessment (QMRA) was determined using a four-step procedure to estimate the risk of response (foodborne illness or death) with respect to a known dose of a pathogen. The hazard identification, exposure assessment, dose-response, and risk characterization were evaluated using the Beta-Poisson model employing Firth's model alongside its correction bias. The QMRA was figuratively expressed from processing to consumption, and the probability of adverse health outcome was evaluated (Schelin et al., \u003cspan citationid=\"CR64\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Pepper et al., \u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e2015\u003c/span\u003e).\u003c/p\u003e \u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003eData Analysis\u003c/h2\u003e \u003cp\u003eAll statistical analyses were conducted using Microsoft Excel 2019 and R statistical programming environment. Descriptive statistics were calculated for all variables. The normality of continuous variables was assessed using Shapiro-Wilk tests. One-way ANOVA followed by Tukey's Honest Significant Difference (HSD) post-hoc test was employed to compare mean bacterial counts across groups, as this test controls for family-wise error rate and is appropriate for multiple comparisons (Nanda et al., \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Kakati \u0026amp; Gogoi, \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). For dose-response modeling, the beta-Poisson model was selected based on its established use for \u003cem\u003eS. aureus\u003c/em\u003e risk assessment (Schelin et al., \u003cspan citationid=\"CR64\" class=\"CitationRef\"\u003e2011\u003c/span\u003e) and its ability to model low-dose foodborne intoxication probabilities. The model parameters (α and β) were derived from selected literatures (Schelin et al., \u003cspan citationid=\"CR64\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Sergelidis et al., \u003cspan citationid=\"CR65\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Schelin et al., \u003cspan citationid=\"CR63\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Pekana \u0026amp; Green, \u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). Firth\u0026rsquo;s penalized logistic regression was employed to reduce small‑sample bias and to handle sparse data and separation in the dose\u0026ndash;response analysis, ensuring finite and more reliable coefficient estimates and confidence intervals (Heinze \u0026amp; Puhr, \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Rahman \u0026amp; Sultana, \u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Walker \u0026amp; Smith, \u003cspan citationid=\"CR71\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). This approach reduces bias in maximum likelihood estimates when event rates are low. Sensitivity analysis varied α and β parameters by \u0026plusmn;\u0026thinsp;20% of their estimated values to assess robustness of intoxication probability estimates. All analyses were conducted in R version 4.5.0 using packages: betareg, and logist. Statistical significance was set at \u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05 (Ogofure et al., \u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Ogofure \u0026amp; Green, \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2025\u003c/span\u003e; Ogofure et al., \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e2025\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e"},{"header":"Results and Discussion","content":"\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003eSociodemographic of Fish Smoking and Vendors in Benin City, Nigeria\u003c/h2\u003e \u003cp\u003eUnless otherwise stated, questionnaire-based analyses are reported for respondents with complete data (\u003cem\u003en\u003c/em\u003e\u0026thinsp;=\u0026thinsp;301), whereas microbiological and quantitative microbial risk assessment analyses were conducted on all smoked fish samples collected (\u003cem\u003en\u003c/em\u003e\u0026thinsp;=\u0026thinsp;330). The social demographic/socio-economic status of the participants or respondents (vendors) in the study is shown in Tables\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e1\u003c/span\u003ea, b and c. Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e1\u003c/span\u003ea. showed that about 44.19% of the respondents were aged 31\u0026ndash;40 years, while 27.91% of the respondents were aged 21\u0026ndash;30 years. All the respondents or fishmongers in the study location were females. About 65.12% of the respondents had primary education as their highest level of education, while 32.56% of the respondents had secondary as their highest level of education (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e1\u003c/span\u003ea). Only about 2.33% of the respondents had polytechnic education as their highest level of education. 79.07% of respondents were in full-time fish-selling businesses in the study location, while 18.6% were part-time fish sellers. Most of the fish sold in the study location were Scomber or Titus, which were sold by 39.87% of the respondents, while Meluza and Herring fish were sold by 33.22% and 18.27% of the respondents, respectively. 90.7% of the respondents participated in the smoking process of the fish sold in the study location, while 9.3% of the respondents did not participate in the smoking process. Similarly, 90.7% of the materials used for smoking the fish were of wood origin, while 4.65% were from wood shavings (sawdust) (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e1\u003c/span\u003ea).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003ea. Sociodemographic/Socioeconomic Status of the Participants (\u003cem\u003en\u003c/em\u003e\u0026thinsp;=\u0026thinsp;301)\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"3\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eVariable\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFrequency\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePercentage\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAge\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e10-20yrs\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2.33\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e21-30yrs\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e84\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e27.91\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e31-40yrs\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e133\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e44.19\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e41-50yrs\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e77\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e25.58\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e51-60yrs\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.00\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eSex\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.00\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFemale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e301\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e100.00\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eLevel of Education\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePrimary\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e196\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e65.12\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSecondary\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e98\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e32.56\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePolytechnic\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2.33\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eDo You Sell Fish as Full Time, Part-Time, or Seasonal Hustle\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFull-time\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e238\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e79.07\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePart-time\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e56\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e18.60\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSeasonal hustle\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2.33\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eType of fish Sold\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSese\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e5.32\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eScomber/Titus\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e120\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e39.87\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMeluza\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e100\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e33.22\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHerring fish\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e55\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e18.27\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCatfish\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e3.32\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eDirectly Participation in the Smoking Process of the Fish\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eYes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e273\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e90.70\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNo\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e9.30\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eMaterial used for smoking the fishes\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eWood smoke\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e273\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e90.70\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSawdust smoke\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e4.65\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOthers\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e4.65\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eFurther socio-demographic characteristics is shown in Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e1\u003c/span\u003eb. Most of the fish samples evaluated in the study were smoked on the same day (65.12%), while less than 30% (27.91%) were smoked a day before the analyses were carried out on the fish. Less than 5% and 3% of the fish were smoked 2 and 3 days before analyses, respectively. About 65.12% of the respondents revealed that they do not have any major preservation for the smoked fish, while 34.88% opined that they use a particular measure of preservation of the smoked fish. Most of the respondents were new to the fish business, as about 30.23% were between 1 and 2 years in the fish business, while about 35.58% had spent more than six years in the fish business (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e1\u003c/span\u003eb). Regardless, most respondents were unaware that the fish could be contaminated with germs and could serve as a reservoir of pathogens to man. This makes up about 65.12% of the respondents who claim to not know about the contamination of fish with germs. The materials used for wrapping the fish, as revealed by the respondents, were basically polythene materials, which 62.9% of the respondents had said they used for wrapping the smoked fish. After smoking the fish, on transit to the market, about 51.16% of the respondents opined that they do not cover the fish between the point of smoking and the point of sale. 48.84% of respondents reveal that they use a covering material for the fish (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e1\u003c/span\u003eb).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eb. Sociodemographic/Socioeconomic Status of the Participants (\u003cem\u003en\u003c/em\u003e\u0026thinsp;=\u0026thinsp;301)\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"3\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eVariable\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFrequency\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePercentage\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHow long has the fish been smoked?\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eToday\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e196\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e65.12\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eYesterday\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e84\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e27.91\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2 days ago\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e4.65\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e3 days ago\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2.33\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMore than 3 days ago\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.00\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eAny measure of preservation for the smoked?\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eYes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e105\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e34.88\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNo\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e196\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e65.12\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eDuration of Participants in the business of selling fish\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1-2yrs\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e91\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e30.23\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e3-4yrs\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e84\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e27.91\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e5-6yrs\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e49\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e16.28\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMore than 6 years\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e77\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e25.58\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eAwareness or knowledge about the contamination of fish with germs\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eYes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e105\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e34.88\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNo\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e196\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e65.12\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eMaterials used for wrapping the smoked fish\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePaper\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e13.95\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCellophane\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e189\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e62.79\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eBoth\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e70\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e23.26\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eAny covering for the fish after smoking in transit to the market?\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eYes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e147\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e48.84\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNo\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e154\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e51.16\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThe socio-demographic/economic status of the participants is also shown in Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e1\u003c/span\u003ec. The socio-demographics of the participants in the study, and concerning the materials used for covering the fish, (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e1\u003c/span\u003ec) revealed that 33.33% of the respondents sometimes wash these materials. In contrast, 28.75% of the respondents do not wash the materials at all, which could also be a source of contamination for the already smoked fish. Most of the fish sellers in the study usually smoke less than a hundred fish to be sold for the day, while 4.65% smoke about 200\u0026ndash;300 fish to be sold for the day. The materials obtained from the materials used in the smoking of the fish, as reported by the respondents, were sourced locally by 18.06%. In comparison, 81% of the respondents said that they purchased the materials for smoking fish, such as cartons, cellophane, etc (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e1\u003c/span\u003ec). The rich tradition of smoking fish in Nigeria reflects not only its practical necessity for food preservation but also its cultural significance, which is deeply embedded in certain local communities.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003ec. Sociodemographic/Socioeconomic Status of the Participants (\u003cem\u003en\u003c/em\u003e\u0026thinsp;=\u0026thinsp;301)\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"3\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eVariable\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFrequency\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePercentage\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFrequency of washing the covering material\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eVery often\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e28.57\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSometimes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e49\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e33.33\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOnce in a while\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e9.52\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNone at all\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e28.57\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eVery often\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e28.57\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eWhen last did Participants wash the covering material?\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eToday\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e63\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e60.00\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eYesterday\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e26.67\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2 days ago\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e13.33\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eNumber of fish smoked and Sold in a batch\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;100 fishes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e231\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e76.74\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e101\u0026ndash;200 fish\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e56\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e18.60\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e201\u0026ndash;300 fish\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e4.65\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u0026gt;\u0026thinsp;300 fish\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.00\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eWhere participants obtain the materials for fish processing\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePurchase them\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e245\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e81.40\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSources for them locally\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e56\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e18.60\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eDo Participants practice cleaning the fish surface with foam?\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eYes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e266\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e88.37\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNo\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e35\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e11.63\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eHow often do participants wash the foam to clean the fish?\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eEveryday\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e91\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e34.21\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTwice weekly\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e112\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e42.11\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOnce a month\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e49\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e18.42\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAnytime\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e5.26\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eHow often do participants make carton material for smoked fish?\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOnce\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e133\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e44.19\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eEvery time\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e126\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e41.86\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOthers\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e13.95\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThe smoking of fish, as an age-old practice, is passed down through generations, and it connects with societal structures and economic dynamics, shaping culinary traditions and livelihoods alike. Exploring the socio-demographic landscape of the smoked fish business exposes a plethora of interconnected factors, which are expressed in this study. Economic conditions play a pivotal role, as the trade of smoked fish often serves as a vital source of livelihood for many households, especially in coastal and riverine communities where fishing is a primary occupation (Adeyeye \u0026amp; Oyewole, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Adeyeye, \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). The socio-demographic and economic aspects of the smoked fish business in Nigeria are multifaceted and influenced by various factors, including cultural practices, economic conditions, and geographical location. Fish smoking is a skill that is learned over time, and older individuals in fishing communities may have extensive experience in traditional smoking methods (Omeje et al., \u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). The results of the socio-economic or demographics of fish mongers in this study were consistent with several reports in the literature about feminine domination of fish processing and smoking activities in many Nigerian communities, and most of these women were reported to have little or no formal education (Ayo-Olalusi et al., \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Omoruyi et al., \u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Adetimehin \u003cem\u003eet al.\u003c/em\u003e, 2019; Oduwale, \u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Ayeloja et al., \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Ayeloja et al., \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Abolagba et al., \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). While it is worthy of note that all fish vendors encountered in the study were females, Oduwale (\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e2019\u003c/span\u003e) had the same statistics. However, at variance with the findings in this study, there are few reports about the involvement of men in the fish business or smoking activity (Ayeloja et al., \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Abolagba et al., \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). While all studies agreed that the demographics of fish selling and smoking activity is largely tilted toward the feminine gender, Abolagba et al. (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2023\u003c/span\u003e) opined that 70% of fish sellers were females while 30% were males. The report, however, varied with the findings in this study as all (100%) of the fish sellers were females. Concerning the education of fish vendors in the study, the results obtained were consistent with the report that about 65.5% (above half the population of fish vendors) had only primary education as their highest qualification. Similarly, findings by Oduwale (\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e2019\u003c/span\u003e) revealed that over 50% of fish vendors in Benin City had Primary education as their highest level of education, while other studies, as in the reports of Adetimehin \u003cem\u003eet al.\u003c/em\u003e (2019) and Ayeloja et al. (\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) revealed that more than 60% (combining no formal education and primary education) of fish vendors either had no formal education or primary education.\u003c/p\u003e \u003cp\u003eFurthermore, the findings in this study concerning the age of fish vendors were consistent with the result obtained by Abolagba et al. (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2023\u003c/span\u003e), who reported that 71% of farmers or sellers were within the age bracket of 25 to 40 years. While fish smoking is a primary occupation for some vendors, others may engage in it as a supplementary source of income (Adetimehin \u003cem\u003eet al.\u003c/em\u003e, 2019; Abolagba et al., \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). The aforementioned reports are consistent with the results obtained in this study, where some participants (fish vendors) claimed that the fish business is more like a side hustle for them, but others engage in the business as their primary source of income or livelihood. In this research, most of the vended smoked fish were Scomber, Hose mackerel and meluza, and this finding was consistent with the report by Oduwale (\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e2019\u003c/span\u003e), who observed that Sese (Horse Mackerel), Scomber (Mackerel) and Meluza (Hake) were the predominant species of smoked fish sold in open markets in Edo State. Additionally, similar to the report mentioned earlier, Abolagba et al. (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2023\u003c/span\u003e) opined that Scomber (\u003cem\u003eScombar scombrus\u003c/em\u003e) and Hake or Meluza (\u003cem\u003eMerluccius merluccius\u003c/em\u003e) were amongst the preferred fish types smoked in Benin in addition to tilapia and catfish however, catfish (\u003cem\u003eClarias gariepinus\u003c/em\u003e) emerged as the preferred species of fish consumed in Benin City, Edo State. The above point by Abolagba et al. (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2023\u003c/span\u003e) about the most preferred fish consumed in Benin City was at variance with the report of Oduwale (\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e2019\u003c/span\u003e), who reported that Scomber and Hake were the most preferred fish sold in open markets in Edo State.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003eIsolation of Coagulase-Positive and Negative Staphylococci from Smoked Fish\u003c/h2\u003e \u003cp\u003eThe socio-demographic characteristics of fish vendors can significantly influence the microbiological quality of smoked fish with dire public health implications. Vendors with limited formal education may lack awareness of essential hygiene practices, increasing the risk of bacterial contamination during processing. Traditional smoking methods, often practiced by vendors, may not adhere to modern food safety standards, allowing pathogens like \u003cem\u003eStaphylococcus aureus\u003c/em\u003e to thrive. The dominance of women in the fish-smoking trade, coupled with harsh economic realities and constraints, can further exacerbate contamination risk, as many vendors lack access to modern equipment or hygienic facilities.\u003c/p\u003e \u003cp\u003eAdditionally, because some vendors engage in fish smoking as a supplementary income source, they might employ some casual approach to hygiene and a plethora of inconsistent processing practices, which might also impact the microbiology and safety of the fish. The combination of these demographics can create an environment where harmful bacteria can proliferate, posing serious public health threats, especially in the form of foodborne illnesses. Therefore, addressing these socio-demographic challenges through better education, improved access to modern technologies, and targeted interventions is critical to safeguarding both food quality and public health. Based on the above premise, the coagulase-positive \u003cem\u003eStaphylococcal\u003c/em\u003e counts in smoked fish samples, considering various socio-demographic and processing factors, revealed observable trends, although most differences were not statistically significant (\u003cem\u003ep\u0026thinsp;\u0026gt;\u0026thinsp;0.05\u003c/em\u003e) (Supplementary Material S2.0 (Figure \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e)). Samples from fish vendors or respondents who were unaware that fish could act as a reservoir of bacterial pathogens exhibited higher \u003cem\u003eStaphylococcal\u003c/em\u003e counts compared to those with knowledge of this fact. However, this difference was not statistically significant (\u003cem\u003ep\u0026thinsp;\u0026gt;\u0026thinsp;0.05\u003c/em\u003e). The counts ranged from 2.56\u0026thinsp;\u0026plusmn;\u0026thinsp;1.48 Log CFU/g in samples from vendors who were aware of the risk to 2.70\u0026thinsp;\u0026plusmn;\u0026thinsp;1.34 Log CFU/g in samples from vendors without such knowledge [Supplementary Material S2.0 (Figure \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e)].\u003c/p\u003e \u003cp\u003eSimilarly, the use of preservative measures was not statistically associated with Staphylococcal load in the smoked fish samples. Although our study found no statistically significant association between reported preservation practices and bacterial load (p\u0026thinsp;\u0026gt;\u0026thinsp;0.05), this finding should not be interpreted as evidence that preservation is unimportant. The absence of statistical significance warrants cautious interpretation, as fewer than 35% of vendors reported using preservation measures, which likely limited statistical power to detect meaningful differences. Furthermore, the study design presents inherent limitations, including a small subgroup sample size, substantial heterogeneity in preservation methods ranging from simple covering to the use of chemical preservatives, and reliance on self-reported practices rather than direct observation, introducing potential measurement error. Vendor awareness was assessed through a single dichotomous question, which may not adequately capture the complexity of food safety knowledge, and residual confounding by unmeasured hygiene practices cannot be excluded. It is well-established that proper preservation and refrigeration are critical for preventing bacterial proliferation in smoked fish (Osibona \u0026amp; Amaechi, \u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Ubadire-Agua et al., \u003cspan citationid=\"CR69\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Huang et al., \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Our findings do not contradict this principle but rather highlight the need for more rigorous assessment of specific preservation methods and their implementation in real-world market settings.\u003c/p\u003e \u003cp\u003eThe bacterial counts ranged from 2.52\u0026thinsp;\u0026plusmn;\u0026thinsp;1.42 Log CFU/g in fish where preservatives were used to 2.64\u0026thinsp;\u0026plusmn;\u0026thinsp;1.46 Log CFU/g in fish without preservatives, indicating that no measurable reduction in bacterial load was observed within the context of this study [Supplementary Material S2.0 (Figure S2)]. When considering the type of smoking material used, no significant difference was observed between fish smoked using wood and those smoked using sawdust [Supplementary Material S2.0 (Figure S3)].\u003c/p\u003e \u003cp\u003eBoth wood and sawdust originate from the same raw material, with sawdust being a byproduct of wood processing in mills. Consequently, their chemical composition, including key antimicrobial components such as phenols, carbonyls, and organic acids, is inherently similar, which likely accounts for the minimal variability observed in their microbial impact. However, both materials were included in our study to explore whether differences in particle size, combustion rate, and heat distribution during the smoking process could influence microbial contamination indirectly. These factors, while subtle, may have practical implications for food safety and quality. The Staphylococcal counts ranged from 2.02\u0026thinsp;\u0026plusmn;\u0026thinsp;0.8 Log CFU/g for wood-smoked fish to 2.66\u0026thinsp;\u0026plusmn;\u0026thinsp;1.3 Log CFU/g for sawdust-smoked fish, suggesting that the choice of smoking material had minimal impact on bacterial contamination [Supplementary material S2 (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e3\u003c/span\u003e), \u003cem\u003ep\u0026thinsp;\u0026gt;\u0026thinsp;0.05\u003c/em\u003e].\u003c/p\u003e \u003cp\u003eIn contrast, a significant difference in \u003cem\u003eStaphylococcal\u003c/em\u003e load was noted based on the type of fish. Specifically, Meluza and Scomber fish exhibited significantly higher Staphylococcal counts compared to Catfish (\u003cem\u003ep\u0026thinsp;\u0026lt;\u0026thinsp;0.05\u003c/em\u003e) [Supplementary Material S2.0 (Figure S4)]. However, no significant difference was found between other fish types, such as Herring and Mackerel. These findings underscore the role of fish type as a potential risk factor for \u003cem\u003eStaphylococcus\u003c/em\u003e contamination in smoked fish products, highlighting the need for targeted interventions based on fish species to improve food safety.\u003c/p\u003e \u003cp\u003eThe duration between smoking and consumption is a critical yet often overlooked factor influencing the microbial quality of smoked fish, particularly for consumers who ingest the product without further cooking. This study evaluated the coagulase-positive \u003cem\u003eStaphylococcal\u003c/em\u003e count of smoked fish samples based on the time elapsed between smoking and analysis, which serves as a proxy for the period before consumption. The \u003cem\u003eStaphylococcal\u003c/em\u003e load varied significantly with the duration between smoking and analysis, ranging from 2.19\u0026thinsp;\u0026plusmn;\u0026thinsp;1.48 Log CFU/g in fish analyzed on the same day of smoking to 3.59\u0026thinsp;\u0026plusmn;\u0026thinsp;0.40 Log CFU/g in fish analyzed three days post-smoking [Supplementary Material S2.0 (Figure S5)]. Statistical analysis revealed a significant increase in \u003cem\u003eStaphylococcal\u003c/em\u003e counts between fish evaluated on the same day of smoking and those evaluated after two and three days (\u003cem\u003ep\u0026thinsp;\u0026lt;\u0026thinsp;0.05\u003c/em\u003e). Conversely, no significant difference was observed between fish analyzed on the same day of smoking and those analyzed the following day, suggesting a significant (\u003cem\u003ep\u0026thinsp;\u0026lt;\u0026thinsp;0.05\u003c/em\u003e) bacterial growth surge occurs after the first 48 h. post-smoking.\u003c/p\u003e \u003cp\u003eThis finding highlights the rapid bacterial proliferation that can occur in smoked fish when left unrefrigerated or improperly stored for extended periods. The over one-log increase in bacterial counts observed after 48 h underscores the heightened risk of Staphylococcal contamination in fish that are consumed long after smoking. Such delays between smoking and consumption could pose serious public health risks, particularly in the context of coagulase-positive \u003cem\u003eStaphylococcus\u003c/em\u003e, which is known to produce enterotoxins capable of causing foodborne illness. These results stress the importance of educating consumers on the risks associated with prolonged storage of smoked fish and emphasize the need for appropriate preservation.\u003c/p\u003e \u003cp\u003eFrom a public health perspective, the higher microbial load in fish consumed days after smoking can elevate the risk of foodborne illnesses. The significant increase in bacterial count over 48 h suggests that smoked fish may become unsafe for consumption within just a few days if not properly stored. This is particularly concerning, especially for individuals consuming the fish without prior or proper processing. The ingestion of fish with high levels of coagulase-positive \u003cem\u003eStaphylococcus\u003c/em\u003e species may lead to food poisoning (Martins et al., \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). Therefore, consumers, especially those in informal markets where cold chain preservation may be limited, face heightened risks of foodborne illness if they consume smoked fish that has been improperly stored or left for several days post-smoking. Public health interventions should emphasize the importance of proper storage and timely consumption of smoked fish to minimize these risks.\u003c/p\u003e \u003cp\u003eThe coagulase-negative staphylococcal count of smoked fish samples based on the use of any preservative measure was found to range from 1.36\u0026thinsp;\u0026plusmn;\u0026thinsp;1.20 Log CFU/g (fishes smoked without any measure of preservation) to 0.78\u0026thinsp;\u0026plusmn;\u0026thinsp;1.04 (fishes smoked with some measure of preservation when not sold for the day) [Supplementary Material S2.0 (Figure S6)]. It was also revealed that there was a statistical difference (\u003cem\u003ep\u0026thinsp;\u0026lt;\u0026thinsp;0.005\u003c/em\u003e) in the coagulase-negative staphylococcal count of smoked fish samples without some measure of preservation compared to fish samples from respondents who used some measure of preservation. The coagulase-negative staphylococcal count of smoked fish samples based on the awareness or knowledge of vendors about fish being a reservoir ranged from 1.77\u0026thinsp;\u0026plusmn;\u0026thinsp;1.34 Log CFU/g (for vendors who know fish being a reservoir) to 1.96\u0026thinsp;\u0026plusmn;\u0026thinsp;1.41 Log CFU/g (for vendors without knowledge of fish being a reservoir) [Supplementary Material S2.0 (Figure S7)]. It was observed that there was no statistical difference in the count of fish obtained from vendors with the awareness of smoked fish being a reservoir of bacteria.\u003c/p\u003e \u003cp\u003eThe coagulase-negative staphylococcal count based on the type of materials used for smoking (wood smoke and sawdust) ranged from 1.52\u0026thinsp;\u0026plusmn;\u0026thinsp;1.11 Log CFU/g (fishes smoked using firewood) to 3.01\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58Log CFU/g (fishes smoked using sawdust) [Supplementary Material S2.0 (Figure S8)]. There was a statistical difference (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05) in the count of smoked fish on the basis of the materials (wood and sawdust) used for smoking or processing of the fish. While coagulase-negative staphylococci are generally less pathogenic, their presence reflects hygiene and handling conditions and may contribute to spoilage and opportunistic risks. The coagulase-negative staphylococcal counts of smoked fish samples based on the fish types evaluated in the study ranged from 1.43\u0026thinsp;\u0026plusmn;\u0026thinsp;1.10 Log CFU/g (Meluza) to 2.40\u0026thinsp;\u0026plusmn;\u0026thinsp;1.51 Log CFU/g (Herring) [Supplementary Material S2.0 (Figure S9)]. Statistically, there was no significant difference (\u003cem\u003ep\u0026thinsp;\u0026gt;\u0026thinsp;0.05\u003c/em\u003e) in the coagulase-negative staphylococcal counts of all the smoked fish samples evaluated in the study. The coagulase-negative staphylococcal counts of smoked fish samples on the basis of the duration of purchase before analysis were found to range from 1.61\u0026thinsp;\u0026plusmn;\u0026thinsp;1.21 Log CFU/g (fishes smoked on the same day of analysis) to 2.16\u0026thinsp;\u0026plusmn;\u0026thinsp;1.87 Log CFU/g (fishes smoked two days before analysis) [Supplementary Material S2.0 (Figure S10)]. It was observed that there was no statistical difference in the coagulase-negative staphylococcal counts of the smoked fish samples based on the duration of smoking before analysis.\u003c/p\u003e \u003cp\u003eIt has been reported that smoked fish, especially at the retail level, might be a potential source of foodborne bacterial pathogens if not properly handled (Ehizibolo et al., \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2008\u003c/span\u003e; Abolagba \u0026amp; Igbinevbo, \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2010\u003c/span\u003e). There are numerous reports in the literature concerning the contamination of smoked fish and ready-to-eat fish sold by fish vendors in local markets around different parts of Nigeria. The isolation of \u003cem\u003eStaphylococcus\u003c/em\u003e from smoked fish (especially in the context of its burdens or estimates) is a critical aspect of food safety and public health, as \u003cem\u003eStaphylococcus\u003c/em\u003e species are known pathogens with a reputation for causing foodborne illnesses (Ogofure \u0026amp; Igbinosa, \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). At the same time, several studies have reported the presence of bacteria in smoked fish samples sold in open markets within Nigeria, only a limited number of studies evaluated and enumerated the burden or population of specific bacteria, such as \u003cem\u003eStaphylococcus\u003c/em\u003e species in smoked fish or ready-to-eat fish. Akinwumi and Adegbehingbe (\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2015\u003c/span\u003e) evaluated the presence of \u003cem\u003eS. aureus\u003c/em\u003e in smoked fish like Tilapia, Herring and Catfish. They opined that the staphylococcal count ranged from 1.4 \u0026times; 10\u003csup\u003e4\u003c/sup\u003e Log CFU/g \u0026minus;\u0026thinsp;2.8 \u0026times; 10\u003csup\u003e5\u003c/sup\u003e Log CFU/g and these counts were a product of poor handling practices and unsafe or unhygienic conditions in which the fish was smoked and exposed to the market. Edeh et al. (\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2023\u003c/span\u003e) also isolated \u003cem\u003eStaphylococcus aureus\u003c/em\u003e from smoked fish from three different Nigerian markets in South-Eastern Nigeria, but they never quantified the staphylococcal burden of the evaluated samples; rather, their analyses were more of a qualitative evaluation. Jega et al. (\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2019\u003c/span\u003e) evaluated the microbiological burden of smoked fish sold at an open market in the Northern part of Nigeria and revealed that a certain sample had a Staphylococcal count of 5.4 \u0026times; 10\u003csup\u003e5\u003c/sup\u003e Log CFU/g. They argued that a multitude of factors could be responsible for the poor microbiological quality of the smoked fish, which include the intrinsic environment where the fish was obtained before smoking, as well as the postharvest handling and processing methods employed by the sellers or vendors. Concerning the handling practices of fish vendors, Christoper et al. (\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2022\u003c/span\u003e) reported that the handling practices and hygienic conditions of smoked fish sold in open markets in Ado-Ekiti were critical to the presence of pathogenic \u003cem\u003eStaphylococcus aureus\u003c/em\u003e in the smoked fish. It is worthy of note that the staphylococcal burden evaluated for both coagulase-positive and negative species in this study was found to be 2-log lesser than the aforementioned studies. Based on findings from this study, vendors are highly encouraged to imbibe simple handwashing practices with soap and clean water, make use of affordable and locally available disinfectants like vinegar or sanitizers for cleaning surfaces and equipment (Ogofure et al., \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e2018\u003c/span\u003e), and potentially adopt heat-based sterilization methods (e.g., boiling utensils). Additionally, the importance of basic hygiene training for vendors to increase awareness and encourage adherence to these practices cannot be overstated. Although the smoked fish samples in this study were not vacuum-packed, the observed low levels of \u003cem\u003eS. aureus\u003c/em\u003e and coagulase-negative staphylococci (\u0026lt;\u0026thinsp;4 log CFU/g) align with previous findings in cold-smoked, vacuum-packed trout and salmon (Gonzalez-Rodriguez et al., \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2002\u003c/span\u003e). However, caution should be exercised in interpreting these comparisons due to differences in packaging and storage conditions. The high salinity of the water in which the salmons were raised likely inhibited the growth of other competing bacteria, thereby creating conditions that favoured the proliferation of halophilic \u003cem\u003eStaphylococcus\u003c/em\u003e species. However, the findings by Prasad et al. (\u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e2007\u003c/span\u003e) reported an average of 4.35\u0026ndash;4.78 log Log CFU/g of total staphylococci in freshwater prawns, and this value was beyond values obtained in this study and also the permissible levels or limit for staphylococci in food. A few of the samples obtained in this study were found to be greater than a 2-log count (in terms of staphylococcal density) with respect to the permissible limits of Staphylococci in foods or smoked fish. Bergdoll (1979), as cited by Prasad et al. (\u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e2007\u003c/span\u003e), suggested that staphylococci count greater than 2-log Log CFU/g is a potential health hazard, and such food should be considered unsafe for consumption (although regulatory limits vary by countries and food products). This implies that going by the counts obtained in this study, most of the smoked fish samples, especially those smoked a day and 2 days before analyses, were above the permissible limit of log 2 Log CFU/g and thus unfit for consumption. Another study by Onjong et al. (\u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) simulated the levels of \u003cem\u003eS. aureus\u003c/em\u003e on slated, sun-dried, and smoked Nile tilapia fish. They revealed that the staphylococcal burden ranged from 4.0 log Log CFU/g to 9.01 log Log CFU/g. The presence of \u003cem\u003eStaphylococcus\u003c/em\u003e species in smoked fish is expected because the bacteria can survive and grow in environments with low water activity and even in environments with high salt concentrations (Gonzalez-Rodriguez et al., \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Prasad et al., \u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e2007\u003c/span\u003e). Similar to the findings in this study, where both coagulase-positive and coagulase-negative \u003cem\u003eStaphylococcus\u003c/em\u003e isolates were detected in smoked fish, Hammad et al. (\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2012\u003c/span\u003e) reported comparable results from fish sold in grocery stores. Their study identified not only \u003cem\u003eStaphylococcus aureus\u003c/em\u003e contamination but also pathogenic strains of coagulase-positive staphylococci, coagulase-negative staphylococci, methicillin-resistant staphylococci, and methicillin-resistant coagulase-negative staphylococci. These observations underscore the widespread prevalence of both coagulase-positive and coagulase-negative staphylococcal species in various fish products, highlighting significant food safety concerns and the potential for antibiotic-resistant strains to pose additional public health risks. Most of the reports made use of mannitol salt agar, which has long been employed for isolating staphylococci, as described by Bridson (\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2006\u003c/span\u003e). Previous studies have expanded traditional mannitol salt agar workflows by incorporating additional selective media for isolation of presumptive pathogenic staphylococci (Ogofure \u0026amp; Igbinosa, \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Mues et al., \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2025\u003c/span\u003e). They included Vogel Johnson agar (which differentiates coagulase-positive \u003cem\u003eStaphylococcus aureus\u003c/em\u003e) and oxacillin-resistant screening agar to identify methicillin-resistant staphylococci. This multi-faceted approach enhanced the precision of isolating and characterizing staphylococcal species, particularly in differentiating antibiotic-resistant strains, offering a more comprehensive tool for staphylococcal detection in food microbiology contexts. In this study, we decided to increase the salinity of the saline solution to 7.5% w/v to achieve more precise isolation of halophilic \u003cem\u003eStaphylococcus\u003c/em\u003e species while employing the sequence of the three-differential media. This elevated salinity level was intended to selectively promote the growth of halotolerant staphylococci while discouraging the proliferation of other non-halophilic microbial isolates. This modification further refined the isolation process, ensuring a more targeted and accurate assessment of staphylococcal contamination in smoked fish.\u003c/p\u003e \u003cp\u003eThe prevalence of various \u003cem\u003eStaphylococcal\u003c/em\u003e isolates from smoked fish is shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. Remarkably, 97.78% of the smoked fish samples were found to be contaminated with \u003cem\u003eStaphylococcus\u003c/em\u003e species. Among these, 46.67% were contaminated with coagulase-positive \u003cem\u003eStaphylococcus aureus\u003c/em\u003e, while 64.44% harboured coagulase-negative \u003cem\u003eStaphylococcus\u003c/em\u003e species. Furthermore, 37.78% of the samples were contaminated with methicillin-resistant \u003cem\u003eStaphylococcus aureus\u003c/em\u003e (MRSA) (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e.).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eThese findings underscore the widespread contamination of smoked fish with both pathogenic and antimicrobial-resistant \u003cem\u003eStaphylococcal\u003c/em\u003e species, highlighting potential public health risks associated with the consumption of improperly handled or processed fish.\u003c/p\u003e \u003cp\u003eThe prevalence of \u003cem\u003eStaphylococcus\u003c/em\u003e species has been widely reported in the literature, with significant variations across different studies. Hammad et al. (\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2012\u003c/span\u003e) observed a notably comparable higher prevalence of \u003cem\u003eStaphylococcus\u003c/em\u003e species (87%), including pathogenic \u003cem\u003eS. aureus\u003c/em\u003e and methicillin-resistant \u003cem\u003eStaphylococcus aureus\u003c/em\u003e (MRSA), in smoked fish samples. These findings differ considerably from the results in our study, where the prevalence of coagulase-positive \u003cem\u003eS. aureus\u003c/em\u003e and MRSA was recorded at 46.67% and 37.78%, respectively. Similarly, Grigoryan et al. (\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2010\u003c/span\u003e) reported a higher prevalence of \u003cem\u003eS. aureus\u003c/em\u003e (74%) during cold-smoked fish processing, identifying the brining stage as a critical control point for preventing contamination. In contrast to the findings in our study, Vazquez-Sanchez et al. (\u003cspan citationid=\"CR70\" class=\"CitationRef\"\u003e2012\u003c/span\u003e) found a lower prevalence of \u003cem\u003eS. aureus\u003c/em\u003e in smoked fish samples at 26%, with higher contamination levels observed in fresh and frozen products. Similar to the aforementioned study, Pekana and Green (\u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e2018\u003c/span\u003e) observed a lower prevalence rate (less than 21%) of \u003cem\u003eStaphylococcus\u003c/em\u003e species from frozen meat products in South Africa. This may suggest that improved regulatory oversight could mitigate similar contamination risks in Benin City. Furthermore, a study by Karimela et al. (\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2017\u003c/span\u003e) highlighted that \u003cem\u003eS. aureus\u003c/em\u003e was a dominant contaminant in traditionally smoked scad fish from Sangihe Island, with a prevalence of 62%. These comparative findings emphasize the variability in \u003cem\u003eStaphylococcus\u003c/em\u003e contamination across different smoking practices and processing stages, underscoring the need for stringent control measures to reduce the risk of contamination in smoked fish products.\u003c/p\u003e \u003cp\u003e \u003cb\u003eMicrobial Risk Assessment of Smoked Fish Contaminated with\u003c/b\u003e \u003cb\u003eStaphylococcus\u003c/b\u003e \u003cb\u003especies\u003c/b\u003e\u003c/p\u003e \u003cp\u003eThe quantitative microbial risk assessment (QMRA) of \u003cem\u003eStaphylococcus\u003c/em\u003e species in smoked fish, providing a critical evaluation of the risk posed by Staphylococcal contamination to consumers, is shown in Fig.\u0026nbsp;2. QMRA typically follows a four-step framework, hazard identification, hazard characterization, exposure assessment, and risk characterization, each of which was employed in this study to estimate the probability of adverse health outcomes following the consumption of smoked fish contaminated with \u003cem\u003eStaphylococcus\u003c/em\u003e species. In our assessment, the \u0026szlig;-Poisson model was used to characterize the dose-response relationship, which quantifies the probability of adverse health outcome based on the level of bacterial exposure. Our study found a prevalence of Staphylococcal isolates in 97.8% of all smoked fish samples, with 46.67% of these isolates identified as \u003cem\u003eStaphylococcus aureus\u003c/em\u003e post-smoking (Fig.\u0026nbsp;2).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eThe beta-Poisson model describes the probability of adverse health outcome (\u003cem\u003eP\u003c/em\u003e) as a function of the dose (\u003cem\u003eD\u003c/em\u003e) of a pathogen. The model equation is given by:\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\:P=1-{e}^{-(\\beta\\:.D)\\alpha\\:}\\)\u003c/span\u003e\u003c/span\u003e\u003c/p\u003e \u003cp\u003e \u003cb\u003eFigure 2. Graphical representation of quantitative microbial risk assessment of staphylococcal isolates from smoked fish\u003c/b\u003e \u003c/p\u003e \u003cp\u003eMost smoked fish are usually purchased within three days of smoking and are sometimes consumed without further processing, significantly increasing the risk of bacterial transmission, as reported by our earlier findings.\u003c/p\u003e \u003cp\u003eThe probability of adverse health outcome relative to doses of staphylococcal isolates consumed from smoked fish is shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e3\u003c/span\u003e. Utilizing Firth's model to predict the probability of adverse health outcome, it was observed that there is a high likelihood of infection, exceeding 90%, when consumers are exposed to higher doses of \u003cem\u003eStaphylococcus aureus\u003c/em\u003e in smoked fish (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eThis risk is particularly pronounced for individuals consuming fish that has been smoked and left for 2\u0026ndash;4 days before being sold (particularly those purchased greater than 2 days post-smoking). The model suggests that the risk of infection remains alarmingly high, even exceeding 80%, under typical storage and consumption conditions.\u003c/p\u003e \u003cp\u003eThese findings emphasize the critical need for improved handling, storage, and consumer awareness to mitigate the risk of staphylococcal infection. Consumer awareness can be enhanced through community outreach programs, media campaigns utilizing accessible platforms like radio and social media, and collaborations with vendors to educate consumers about proper storage practices, the dangers of consuming contaminated smoked fish, and how to identify spoiled products. The prolonged shelf life of smoked fish, particularly under non-refrigerated conditions, significantly increases the probability of bacterial growth and the subsequent risk of foodborne illness. In essence, while smoking helps preserve fish, it doesn't make it completely safe for extended periods, especially without proper refrigeration. The longer smoked fish is kept, particularly at room temperature, the higher the risk becomes of it harbouring harmful bacteria, in this case, \u003cem\u003eStaphylococcus\u003c/em\u003e species. This underscores the necessity of immediate consumption or proper refrigeration practices to reduce the public health threat posed by \u003cem\u003eStaphylococcus aureus\u003c/em\u003e in smoked fish product.\u003c/p\u003e \u003cp\u003eThe possibility of staphylococcal contamination of smoked fish is a serious threat to public health even though Sikorski and Kalodziejska (\u003cspan citationid=\"CR67\" class=\"CitationRef\"\u003e2002\u003c/span\u003e) asserted that contamination of smoked seafood by \u003cem\u003eStaphylococcus aureus\u003c/em\u003e is rare. Recent studies have increasingly highlighted the issue of \u003cem\u003eStaphylococcus\u003c/em\u003e contamination in food products, including smoked fish. Several researches such as Christoper et al. (\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2022\u003c/span\u003e), Edeh et al. (\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2023\u003c/span\u003e), and Musa et al. (\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e2020\u003c/span\u003e) reported that smoked fish can indeed become contaminated, sometimes with astronomical bacterial burdens. These findings underscore the need for heightened vigilance in food safety practices, particularly regarding smoked fish, as the risk of contamination appears to be rising with an increasing number of days post-smoking. Despite these possibilities of Staphylococcal contamination, poor processing and handling of raw and smoked fish can lead to contamination by \u003cem\u003eStaphylococcus\u003c/em\u003e species, which in turn can favour toxin production, increasing the risk of infection in consumers. The vulnerability of smoked fish to \u003cem\u003eStaphylococcus\u003c/em\u003e contamination has been widely reported, highlighting the necessity for a comprehensive risk assessment.\u003c/p\u003e \u003cp\u003eOur study demonstrated that the risk characterization and probability of illness from the consumption of smoked fish contaminated with \u003cem\u003eStaphylococcus\u003c/em\u003e species is dose-dependent. A significant correlation was found between the \u003cem\u003eStaphylococcus\u003c/em\u003e load and various factors, particularly the time elapsed between smoking and consumption. Fish smoked two days post-purchase were observed to harbour a higher microbial load compared to freshly smoked samples, thereby increasing the dose per serving and the subsequent risk of infection. This observation aligns with the findings of Onjong et al. (\u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e2021\u003c/span\u003e), who reported an elevated risk of staphylococcal enterotoxin production in smoked Nile tilapia, particularly in unregulated value chains in Kenya. Their study noted that the time taken to sell fish, whether under ambient or refrigerated conditions, directly influenced the dose-per-serving and enterotoxin production. This parallels our findings, where Staphylococcal load increased with duration post-smoking from 24 to 72 h, and the time spent selling the product contributed further to the risk of toxin production. Additionally, Musa et al. (\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e2020\u003c/span\u003e) identified \u003cem\u003eStaphylococcus aureus\u003c/em\u003e contamination in smoked fish sold at Ahmadu Bello University, underscoring the health risks posed to consumers. Sergelidis et al. (\u003cspan citationid=\"CR65\" class=\"CitationRef\"\u003e2014\u003c/span\u003e) reported that 43% of ready-to-eat fish products, including smoked varieties, present a potential health hazard due to the presence of methicillin-resistant \u003cem\u003eStaphylococcus aureus\u003c/em\u003e (MRSA).\u003c/p\u003e \u003cp\u003eTo mitigate the risk of \u003cem\u003eStaphylococcus\u003c/em\u003e contamination and toxin production, strict adherence to hygiene and best practices during both the smoking and vending processes is critical. Vendors should employ better preservation techniques for unsold fish to prevent microbial proliferation. Furthermore, consumer awareness programs are essential to prevent the possibility of \u003cem\u003eStaphylococcal\u003c/em\u003e food poisoning outbreaks.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003eStudy Limitations\u003c/h2\u003e \u003cp\u003eThis study has some limitations that should be considered when interpreting the findings. Although 330 smoked fish samples were analyzed microbiologically, questionnaire-based analyses were limited to respondents with complete data, which may have reduced statistical power for some comparisons. In addition, information on handling and preservation practices was self-reported and may be subject to recall or social desirability bias. The cross-sectional design also limits causal inference between vendor practices and microbial contamination levels. Nevertheless, the combined use of microbiological analyses and quantitative microbial risk assessment provides a robust, context-specific evaluation of public health risks associated with smoked fish sold in informal markets.\u003c/p\u003e \u003c/div\u003e"},{"header":"Conclusion and Recommendation","content":" \u003cdiv id=\"Sec15\" class=\"Section2\"\u003e \u003cp\u003eThis study provides the first comprehensive quantitative microbial risk assessment of \u003cem\u003eS. aureus\u003c/em\u003e in artisanally smoked fish from randomly selected markets in Edo South senatorial district, revealing alarmingly high MRSA prevalence (37.78%) and demonstrating critical time-dependent contamination thresholds. While bacterial proliferation in unrefrigerated food is well-established, our study uniquely quantifies infection probability at specific consumption timepoints (\u0026gt;\u0026thinsp;90% risk at 72 h post-smoking) and identifies fish species-specific vulnerability patterns not previously documented in this context. These estimates are model-based and reflect worst-case exposure scenarios under ambient storage conditions. These results reinforce the urgent need for public health efforts, which focus on introducing low-cost refrigeration or insulated storage solutions for vendors, alongside training on hygienic handling and storage methods and increased consumer awareness to minimize the risks associated with smoked fish. More so, immediate consumption or proper refrigeration should be emphasized to reduce the potential for bacterial contamination, particularly with coagulase-positive and methicillin-resistant strains. Our study contributes significantly to UN Sustainable Development Goals (SDGs) 2 (Zero Hunger), 3 (Good Health and Well-being), and 12 (Responsible Consumption and Production). By assessing microbial risks and processing practices of smoked fish, our study directly addresses food safety concerns (SDG 2.1: ensuring access to safe and nutritious food) and promotes public health (SDG 3.3: combating communicable diseases). The identification of critical factors influencing staphylococcal contamination in smoked fish supports the development of sustainable food production systems (SDG 2.4) and encourages responsible consumption patterns (SDG 12.3: reducing food losses along production and supply chains). Our findings on the high prevalence of antibiotic-resistant strains (MRSA) contribute to the global effort to combat antimicrobial resistance (SDG 3.d). By emphasizing the need for enhanced consumer education, improved preservation practices, and stricter regulations, this research supports the implementation of sustainable consumption and production patterns (SDG 12.8: ensuring awareness for sustainable development). Ultimately, these scientific observations provide crucial insights for policymakers, food producers, and consumers to make informed decisions that enhance food safety, reduce health risks, and promote sustainable practices in the fish-smoking industry.\u003c/p\u003e \u003c/div\u003e"},{"header":"Abbreviations","content":"\u003cdiv class=\"DefinitionList\"\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eQMRA\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eQuantitative Microbial Risk Assessment\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eMRSA\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003emethicillin-resistant Staphylococcus aureus\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eSDGs\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eSustainable Development Goals\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eSFP\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eStaphylococcal food poisoning\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eWHO\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eWorld Health Organization\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eLGA\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eLocal Government Areas\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eHSD\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eHonest Significant Difference\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eCoNS\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eCoagulase Negative Staphylococci\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003c/div\u003e"},{"header":"Declarations","content":"\u003cp\u003e \u003ch2\u003eConflict of Interest\u003c/h2\u003e \u003cp\u003eThe authors have no relevant financial or non-financial interests to disclose.\u003c/p\u003e \u003c/p\u003e\u003ch2\u003eFunding Statement\u003c/h2\u003e \u003cp\u003eThis research was funded by the University of Johannesburg University Research Committee, Project No. 075432.\u003c/p\u003e\u003ch2\u003eAuthors\u0026rsquo; contributions\u003c/h2\u003e \u003cp\u003eProgress E. Boyo-Agbonile, Abraham G. Ogofure, Brenda O. Isichel-Ukah, Ezekiel Green and Etinosa O. Igbinosa contributed to the study conception and design. Progress E. Boyo-Agbonile, Abraham G. Ogofure, Brenda O. Isichel-Ukah and Etinosa O. Igbinosa performed material preparation, data collection and analysis. Progress E. Boyo-Agbonile, Abraham G. Ogofure, and Etinosa O. Igbinosa, wrote the first draft of the manuscript, and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.\u003c/p\u003e\u003ch2\u003eAcknowledgements\u003c/h2\u003e \u003cp\u003eThe authors are grateful for the material support from the Department of Biotechnology and Food Technology, University of Johannesburg, South Africa, toward the success of this research. We thank the Alexander von Humboldt Foundation for the equipment grant awarded to Prof. E.O. Igbinosa. More importantly, authors are grateful to God for directing our steps in this research, knowing that \u003cem\u003e\u0026ldquo;it is God who works in us to will and to act\u0026rdquo;\u003c/em\u003e (Philippians 2:13).\u003c/p\u003e\u003ch2\u003eData Availability Statement\u003c/h2\u003e \u003cp\u003eThe data underlying this manuscript are available in the manuscript and in its supplementary material.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAbdi AM, Amano A, Abrahim A, Getahun M, Ababor S, Kumie A (2020) Food Hygiene Practices and Associated Factors Among Food Handlers Working in Food Establishments in the Bole Sub City, Addis Ababa, Ethiopia. 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Toxins (Basel) 8(7). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/toxins8070176\u003c/span\u003e\u003cspan address=\"10.3390/toxins8070176\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":true,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"annals-of-microbiology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"amoa","sideBox":"Learn more about [Annals of Microbiology](https://www.springer.com/journal/13213)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/amoa/default.aspx","title":"Annals of Microbiology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Staphylococcus spp., Quantitative Microbial Risk Assessment (QMRA), Beta-Poisson model, Smoked fish, Food safety, Processing practices, MRSA","lastPublishedDoi":"10.21203/rs.3.rs-8482256/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8482256/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003ePurpose\u003c/h2\u003e \u003cp\u003eStaphylococcal contamination of smoked fish represents a public health concern, particularly in informal food markets. This study investigated the microbial risk associated with \u003cem\u003eStaphylococcus\u003c/em\u003e-contaminated smoked fish, processing practices, and potential public health implications in Benin City, Nigeria. Structured questionnaires were administered to smoked fish vendors to document processing, handling, and storage practices.\u003c/p\u003e\u003ch2\u003eMaterials and Methods\u003c/h2\u003e \u003cp\u003eSmoked fish samples (Scomber, Meluza, Sese, catfish, and herring) were collected and analyzed using standard culture-based and molecular techniques. Questionnaire data were analyzed using chi-square and correlation analyses. Quantitative Microbial Risk Assessment (QMRA) was conducted to estimate the probability of adverse health outcomes following consumption of contaminated fish, using the beta-Poisson dose\u0026ndash;response model.\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e \u003cp\u003eThe results showed that 98% of the smoked fish samples were contaminated with \u003cem\u003eStaphylococcus\u003c/em\u003e species, including coagulase-positive strains. Methicillin-resistant \u003cem\u003eStaphylococcus aureus\u003c/em\u003e (MRSA) was detected in 37.78% of the samples. Fish stored for more than two days post-smoking exhibited significantly higher bacterial counts (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05), with marked logarithmic increases observed within 24\u0026ndash;48 h after smoking. Scomber and Meluza samples had significantly higher staphylococcal loads compared with other fish species. The use of preservatives, vendors\u0026rsquo; awareness of bacterial contamination, and the type of smoking material (wood or sawdust) did not significantly (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05) influence bacterial loads. QMRA outcomes indicated a high predicted probability (\u0026gt;\u0026thinsp;90%) of adverse health effects under ambient storage conditions over 2\u0026ndash;4 d.\u003c/p\u003e\u003ch2\u003eConclusion\u003c/h2\u003e \u003cp\u003eThese findings underscore the need for improved hygiene practices, enhanced consumer and vendor education, better preservation strategies, and stricter regulatory oversight to reduce the risk of staphylococcal foodborne illnesses associated with smoked fish consumption.\u003c/p\u003e","manuscriptTitle":"Microbial risk assessment of Staphylococcus species in smoked fish from informal markets in Edo State, Nigeria: processing practices and public health implications","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-01-22 20:25:05","doi":"10.21203/rs.3.rs-8482256/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"reviewerAgreed","content":"","date":"2026-01-20T11:52:05+00:00","index":0,"fulltext":""},{"type":"reviewersInvited","content":"","date":"2026-01-20T11:24:14+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2026-01-05T13:58:34+00:00","index":"","fulltext":""},{"type":"submitted","content":"Annals of Microbiology","date":"2026-01-02T09:41:21+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"annals-of-microbiology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"amoa","sideBox":"Learn more about [Annals of Microbiology](https://www.springer.com/journal/13213)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/amoa/default.aspx","title":"Annals of Microbiology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"f236d612-99c2-4922-a40b-83c67ac536da","owner":[],"postedDate":"January 22nd, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2026-05-04T16:04:36+00:00","versionOfRecord":{"articleIdentity":"rs-8482256","link":"https://doi.org/10.1186/s13213-026-01858-6","journal":{"identity":"annals-of-microbiology","isVorOnly":false,"title":"Annals of Microbiology"},"publishedOn":"2026-05-01 15:58:05","publishedOnDateReadable":"May 1st, 2026"},"versionCreatedAt":"2026-01-22 20:25:05","video":"","vorDoi":"10.1186/s13213-026-01858-6","vorDoiUrl":"https://doi.org/10.1186/s13213-026-01858-6","workflowStages":[]},"version":"v1","identity":"rs-8482256","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8482256","identity":"rs-8482256","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}