Search results for: T. Shiga

Authors: Mohamed Al-Hazmi, Abdullah Al-Arfaj, Moussa Ihab

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Raw, unpasteurized milk can carry dangerous bacteria such as Salmonella, E. coli, and Listeria, which are responsible for causing numerous foodborne illnesses. The objective of this study was molecular characterization of shiga toxogenic E. coli in raw milk collected from different Egyptian governorates by multiplex PCR. During the period of 25th May to 25th October 2012, a total of 320 bulk-tank milk samples were collected from 10 cow farms located in different Egyptian governorates. Bacteriological examination of milk samples revealed the presence of E. coli organisms in 65 samples (20.3%), serotyping of the E. coli isolates revealed, 35 strains (10.94%) O111, 15 strains (4.69%) O157: H7, 10 strains (3.13%) O128 and 5 strains (1.56%) O119. Multiplex PCR for detection of shiga toxin type 2 and intimin genes revealed positive amplification of 255 bp fragment of shiga toxin type 2 gene and 384 bp fragment of intimin gene from all E. coli serovar O157: H7, while from serovar O111 were 25 (71.43%), 20 (57.14%) and from serovar O128 were 6 (60%), 8 (80%), respectively. The results of multiplex PCR assay are useful for identification of STEC possessing the eaeA and stx2 genes.

Keywords: raw milk, E. coli, multiplex PCR, Shiga toxin type 2, intimin gene

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Authors: Jessica C. Bannon, Cleso M. Jordao Jr., Mohammad Melebari, Carlos Leon-Velarde, Roger Johnson, Keith Warriner

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There has been an increased incidence of non-O157 Shiga Toxin Escherichia coli (STEC) with six serotypes (Top 6) being implicated in causing haemolytic uremic syndrome (HUS). Beef has been suggested to be a significant vehicle for non-O157 STEC although conclusive evidence has yet to be obtained. The following aimed to determine the prevalence of the Top 6 non-O157 STEC in beef processing using three different diagnostic platforms then characterize the recovered isolates. Hide, carcass and environmental swab samples (n = 60) were collected from a beef processing facility over a 12 month period. Enriched samples were screened using Biocontrol GDS, BAX or PALLgene molecular diagnostic tests. Presumptive non-O157 STEC positive samples were confirmed using conventional PCR and serology. STEC was detected by GDS (55% positive), BAX (85% positive), and PALLgene (93%). However, during confirmation testing only 8 of the 60 samples (13%) were found to harbour STEC. Interestingly, the presence of virulence factors in the recovered isolates was unstable and readily lost during subsequent sub-culturing. There is a low prevalence of Top 6 non-O157 STEC associated with beef although other serotypes are encountered. Yet, the instability of the virulence factors in recovered strains would question their clinical relevance.

Keywords: beef, food microbiology, shiga toxin, STEC

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Authors: Jun Hyung Lee, Robin B. Guevarra, Jin Ho Cho, Bo-Ra Kim, Jiwon Shin, Doo Wan Kim, Young Hwa Kim, Minho Song, Hyeun Bum Kim

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Colibacillosis is one of the major health problems in young piglets ultimately resulting in their death, and it is common especially in young piglets. For the swine industry, colibacillosis is one of the important economic burdens. Therefore, it is necessary for the swine industries to prevent Colibacillosis in piglets in order to reduce economic losses. Thus, we tested three types of natural plant extracts (PEs) to evaluate antibacterial effects against Shiga toxin-producing Escherichia coli (STEC) isolated from the piglet. Three PEs including turmeric oleoresin (containing curcumin 79 to 85%), capsicum oleoresin (containing capsaicin 40%-40.1%), and garlic essential oil (100% natural garlic) were tested using the direct contact agar diffusion test, minimum inhibitory concentration test, growth curve assay, and heat stability test. The tests were conducted with PEs at each concentration of 2.5%, 5%, and 10%. For the heat stability test, PEs with 10% concentration were incubated at each 4, 20, 40, 60, 80, and 100 °C for 1 hour, then the direct contact agar diffusion test was used. For the positive and negative controls, 0.5N HCl and 1XPBS were used. All the experiments were duplicated. In the direct contact agar diffusion test, garlic essential oil with 2.5%, 5%, and 10% concentration showed inhibit zones of 1.1cm, 3.0cm, and 3.6 cm in diameters compared to that of 3.5cm diameter for 0.5N HCl. The minimum inhibited concentration of garlic essential oil was 2.5%. Growth curve assay showed that the garlic essential oil was able to inhibit STEC growth significantly after 4 hours. The garlic essential oil retained the ability to inhibit STEC growth after heat treatment at each temperature. However, turmeric and capsicum oleoresins were not able to significantly inhibit STEC growth by all the tests. Even though further tests using the piglets will be required to evaluate effects of garlic essential oil for the Colibacillosis prevention for piglets, our results showed that the garlic essential oil could be used as a potential natural agent to prevent Colibacillosis in swine.

Keywords: garlic essential oil, pig, Colibacillosis, Escherichia coli

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Authors: Igbakura I. Luga, Alex A. Adikwu

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A comparison of resistance to quinolones was carried out on isolates of Shiga toxin-producing Escherichia coliO157:H7 from cattle and mecA and nuc genes harbouring Staphylococcus aureus from pigs. The isolates were separately tested in the first and current decades of the 21^st century. The objective was to demonstrate the dissemination of resistance to this frontline class of antibiotic by bacteria from food animals and bring to the limelight the spread of antibiotic resistance in Nigeria. A total of 10 isolates of the E. coli O157:H7 and 9 of mecA and nuc genes harbouring S. aureus were obtained following isolation, biochemical testing, and serological identification using the Remel Wellcolex E. coli O157:H7 test. Shiga toxin-production screening in the E. coli O157:H7 using the verotoxin E. coli reverse passive latex agglutination (VTEC-RPLA) test; and molecular identification of the mecA and nuc genes in S. aureus. Detection of the mecA and nuc genes were carried out using the protocol by the Danish Technical University (DTU) using the following primers mecA-1:5'-GGGATCATAGCGTCATTATTC-3', mecA-2: 5'-AACGATTGTGACACGATAGCC-3', nuc-1: 5'-TCAGCAAATGCATCACAAACAG-3', nuc-2: 5'-CGTAAATGCACTTGCTTCAGG-3' for the mecA and nuc genes, respectively. The nuc genes confirm the S. aureus isolates and the mecA genes as being methicillin-resistant and so pathogenic to man. The fluoroquinolones used in the antibiotic resistance testing were norfloxacin (10 µg) and ciprofloxacin (5 µg) in the E. coli O157:H7 isolates and ciprofloxacin (5 µg) in the S. aureus isolates. Susceptibility was tested using the disk diffusion method on Muller-Hinton agar. Fluoroquinolone resistance was not detected from isolates of E. coli O157:H7 from cattle. However, 44% (4/9) of the S. aureus were resistant to ciprofloxacin. Resistance of up to 44% in isolates of mecA and nuc genes harbouring S. aureus is a compelling evidence for the rapid spread of antibiotic resistance from bacteria in food animals from Nigeria. Ciprofloxacin is the drug of choice for the treatment of Typhoid fever, therefore widespread resistance to it in pathogenic bacteria is of great public health significance. The study concludes that antibiotic resistance in bacteria from food animals is on the increase in Nigeria. The National Food and Drug Administration and Control (NAFDAC) agency in Nigeria should implement the World Health Organization (WHO) global action plan on antimicrobial resistance. A good starting point can be coordinating the WHO, Office of International Epizootics (OIE), Food and Agricultural Organization (FAO) tripartite draft antimicrobial resistance monitoring and evaluation (M&E) framework in Nigeria.

Keywords: Fluoroquinolone, Nigeria, resistance, Staphylococcus aureus

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Authors: Satoshi Furukawa, Satomu Morita, Katsuji Nishi, Masahito Hitosugi

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We experience the autopsy cases that the deceased was alive in emergency room on arrival. Bleeding is the leading cause of preventable death after injury. This retrospective study aimed to characterize opportunities for performance improvement identified in patients who died from traffic trauma and were considered by the quality improvement of education system. The Japan Advanced Trauma Evaluation and Care (JATEC) education program was introduced in 2002. We focused the abdominal traffic trauma injury. An autopsy-based cross-sectional study conducted. A purposive sampling technique was applied to select the study sample of 41 post-mortems of road traffic accident between April 1999 and March 2014 subjected to medico-legal autopsy at the department of Forensic Medicine, Shiga University of Medical Science. 16 patients (39.0%) were abdominal trauma injury. The mean period of survival after meet with accident was 13.5 hours, compared abdominal trauma death was 27.4 hours longer. In road traffic accidents, the most injured abdominal organs were liver followed by mesentery. We thought delayed treatment was associated with immediate diagnostic imaging, and so expected to expand trauma management examination.

Keywords: abdominal traffic trauma, preventable death, autopsy, emergency medicine

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Authors: Keiko Naito, Y. Nakashima, S. Yamauchi, Y. Kunitani, Y. Ishigami, K. Numata, M. Mizobe, Y. Homma, J. Takahashi, T. Inoue, T. Shiga, H. Funakoshi

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Background: A presence of severe acidemia, defined as pH < 7.2, is common during endotracheal intubation for critically ill patients in the emergency department (ED). Severe acidemia is widely recognized as a predisposing factor for intubation failure. However, it is unclear that acidemic condition itself actually makes endotracheal intubation more difficult. We aimed to evaluate if a presence of severe acidemia before intubation is associated with the lower first attempt intubation success rate in the ED. Methods: This is a retrospective observational cohort study in the ED of an urban hospital in Japan. The collected data included patient demographics, such as age, sex, and body mass index, presence of one or more factors of modified LEMON criteria for predicting difficult intubation, reasons for intubation, blood gas levels, airway equipment, intubation by emergency physician or not, and the use of the rapid sequence intubation technique. Those with any of the following were excluded from the analysis: (1) no blood gas drawn before intubation, (2) cardiopulmonary arrest, and (3) under 18 years of age. The primary outcome was the first attempt intubation success rates between a severe acidemic patients (SA) group and a non-severe acidemic patients (NA) group. Logistic regression analysis was used to test the first attempt success rates for intubations between those two groups. Results: Over 5 years, a total of 486 intubations were performed; 105 in the SA group and 381 in the NA group. The univariate analysis showed that the first attempt intubation success rate was lower in the SA group than in the NA group (71.4% vs 83.5%, p < 0.01). The multivariate logistic regression analysis identified that severe acidemia was significantly associated with the first attempt intubation failure (OR 1.9, 95% CI 1.03-3.68, p = 0.04). Conclusions: A presence of severe acidemia before endotracheal intubation lowers the first attempt intubation success rate in the ED.

Keywords: acidemia, airway management, endotracheal intubation, first-attempt intubation success rate

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Authors: Y. Kunitani, Y. Nakashima, S. Yamauchi, Y. Ishigami, K. Naito, K. Numata, M. Mizobe, Y. Homma, J. Takahashi, T. Inoue, T. Shiga, H. Funakoshi

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Background: Japanese Spotted Fever (JSF) is one of the Rickettsial infections, caused by Rickettsia japonica, which is transmitted by ticks. JSF is seen in limited area, such as Japan and South Korea. Its clinical triad is rash, eschar and fever. It often shows leukocytopenia, thrombopenia, elevated transaminase and high C-reactive protein (CRP). Sometimes it can be life-threatening due to disseminated intravascular coagulation or multiple organ failure. Study Aim: The aim of this study is to describe the features of JSF, as this unique infection is rapidly growing in Japan. Methods: This is a case series of JSF from 2009 to 2016, in Mie Prefectural Hospital in Japan. We collected JSF cases, which were diagnosed by polymerase chain reaction (PCR) of the skin or blood serum, or the elevation of the antibody titer of paired blood samples. Results: There were 43 JSF patients (19 male, 24 female) with a median age of 71 years [IQR:65-80]. The median body temperature was 38.1°C[IQR: 37.5-39.0]. 95% had a rash, 67% had eschar and 50% had fever. The median WBC count was 6,700 [IQR: 5,750-8,200] and leukocytopenia was observed in only 7%. The median platelet count was 14x104 [IQR10x104-17x104], thrombopenia was observed in 65%. The median aspartate transaminase (AST) was 53 IU/L [IQR: 41-93]; the median alanine aminotransferase (ALT) was 34 IU/L [IQR: 24-54]; the median CRP was 10.4 mg/dL [IQR:7.2-13.9]; the median lactate dehydrogenase (LDH) was 352IU/L [IQR:282-451]. CRP and LDH were elevated in almost all of the patients. Median length of stay in hospital was 8 days [IQR: 6-11]. All patients were treated with tetracycline and quinolone on the day of the presentation. There was no fatality from JSF. Conclusion: The patients with JSF classically presents with eschar, rash and fever. However, in this study, the half of the patients were afebrile. Although JSF is not a common infectious disease worldwide, if the patient had previously visited Japan or South Korea and presented with rash and eschar with or without fever, we should consider JSF as a potential diagnosis.

Keywords: infectious disease, Japanese spotted fever, Rickettsial disease, Rickettsia japonica

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Authors: Raana Babadi Fathipour

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The production of dairy holds significant importance in the sustenance of billions of individuals worldwide, as they rely on milk and its derived products for daily consumption. In addition to being a source of essential nutrients crucial for human well-being, such as proteins, fats, vitamins, and minerals; dairy items are witnessing an increasing demand worldwide. Amongst all the factors contributing to the quality and safety assurance of dairy products, the strong focus lies on maintaining high standards in raw milk procurement. Raw milk serves as an externally nutritious medium for various microorganisms due to its inherent properties. This poses a considerable challenge for the dairy industry in ensuring that microbial contamination is minimized throughout every stage of the value chain. Despite implementing diverse process technologies—both conventional and innovative—the occurrence of microbial spoilage still results in substantial losses within this industry context. Moreover, milk and dairy products have been associated with numerous cases of foodborne illnesses across the globe. Various pathogens such as Salmonella serovars, Campylobacter spp., Shiga toxin-producing Escherichia coli, Listeria monocytogenes, and enterotoxin producing Staphylococcus aureus are commonly identified as the culprits behind these outbreaks in the dairy industry. The effective management of food safety within this sector necessitates a proactive and risk-based approach to reform. However, this strategy presents difficulties for developing nations where informal value chains dominate the dairy sector. Whether operating on a small or large scale or falling within formal or informal realms, it is imperative that the dairy industry adheres to principles of good hygiene practices and good manufacturing practices. Additionally, identifying and managing potential sources of contamination is crucial in mitigating challenges pertaining to quality and safety precautions.

Keywords: dairy value chain, microbial contamination, food safety, hygiene

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Authors: Danielle Katrina Santos, Koji Shimada

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Environmentally-friendly agriculture has been promoted for over two decades as a response to the environmental challenges brought by climate change and biological loss. Located above the equator, it is possible that Japan may benefit from future climate change, yet Japan is also a rarely developed country located in the Asian Monsoon climate region, making it vulnerable to the impacts of climate change. In this regard, the Japanese government has initiated policies to adapt to the adverse effects of climate change through the promotion and popularization of environmentally-friendly farming practices. This study aims to determine profit efficiency among environmentally-friendly rice farmers in Shiga Prefecture using the Stochastic Frontier Approach. A cross-sectional survey was conducted among 66 farmers from top rice-producing cities through a structured questionnaire. Results showed that the gross farm income of environmentally-friendly rice farmers was higher by JPY 316,223.00/ha. Production costs were also found to be higher among environmentally-friendly rice farmers, especially on labor costs, which accounted for 32% of the total rice production cost. The resulting net farm income of environmentally-friendly rice farmers was only higher by JPY 18,044/ha. Results from the stochastic frontier analysis further showed that the profit efficiency of conventional farmers was only 69% as compared to environmentally-friendly rice farmers who had a profit efficiency of 76%. Furthermore, environmentally-friendly agriculture participation, other types of subsidy, educational level, and farm size were significant factors positively influencing profit efficiency. The study concluded that substitution of environmentally-friendly agriculture for conventional rice farming would result in an increased profit efficiency due to the direct payment subsidy and price premium received. The direct government policies that would strengthen the popularization of environmentally-friendly agriculture to increase the production of environmentally-friendly products and reduce pollution load to the Lake Biwa ecosystem.

Keywords: profit efficiency, environmentally-friendly agriculture, rice farmers, direct payment subsidies

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Authors: Debabrata Biswas

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The current trend in the development of antibiotic resistance by multiple bacterial pathogens has resulted in a troubling loss of effective antibiotic options for human. The emergence of multi-drug-resistant pathogens has necessitated higher dosages and combinations of multiple antibiotics, further exacerbating the problem of antibiotic resistance. Zoonotic bacterial pathogens, such as Salmonella, Campylobacter, Shiga toxin-producing Escherichia coli (such as enterohaemorrhagic E. coli or EHEC), and Listeria are the most common and predominant foodborne enteric infectious agents. It was observed that these pathogens gained/developed their ability to survive in the presence of antibiotics either in farm animal gut or farm environment and researchers believe that therapeutic and sub-therapeutic antibiotic use in farm animal production might play an important role in it. The mechanism of action of antimicrobial components used in farm animal production in genomic interplay in the gut and farm environment, has not been fully characterized. Even the risk of promoting the exchange of mobile genetic elements between microbes specifically pathogens needs to be evaluated in depth, to ensure sustainable farm animal production, safety of our food and to mitigate/limit the enteric infection with multiple antibiotic resistant bacterial pathogens. Due to the consumer’s demand and considering the current emerging situation, many countries are in process to withdraw antibiotic use in farm animal production. Before withdrawing use of the sub-therapeutic antibiotic or restricting the use of therapeutic antibiotics in farm animal production, it is essential to find alternative natural antimicrobials for promoting the growth of farm animal and/or treating animal diseases. Further, it is also necessary to consider whether that compound(s) has the potential to trigger the acquisition or loss of genetic materials in zoonotic and any other bacterial pathogens. Development of alternative therapeutic and sub-therapeutic antimicrobials for farm animal production and food processing and preservation and their effective implementation for sustainable strategies for farm animal production as well as the possible risk for horizontal gene transfer in major enteric pathogens will be focus in the study.

Keywords: food safety, natural antimicrobial, sustainable farming, antibiotic resistance

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Authors: Deog-Gyu Seo, Jin-Soo Ahn

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Objectives: Recently, the development of adhesive primers on stable bonding between zirconia and resin cement has been on the increase. The bond strength of zirconia-resin cement can be effectively increased with the treatment of primer composed of the adhesive monomer that can chemically bond with the oxide layer, which forms on the surface of zirconia. 10-methacryloyloxydecyl dihydrogen phosphate (10-MDP) that contains phosphate ester and acidic monomer 4-methacryloxyethyl trimellitic anhydride(4-META) have been suggested as monomers that can form chemical bond with the surface oxide layer of zirconia. Also, these suggested monomers have proved to be effective zirconia surface treatment for bonding to resin cement. The purpose of this study is to evaluate the effects of primer treatment on the bond strength of Zirconia-resin cement by using three different kinds of primers on the market. Methods: Zirconia blocks were prepared into 60 disk-shaped specimens by using a diamond saw. Specimens were divided into four different groups: first three groups were treated with zirconiaLiner(Sun Medical Co., Ltd., Furutaka-cho, Moriyama, Shiga, Japan), Alloy primer (Kuraray Noritake Dental Inc., Sakaju, Kurashiki, Okayama, Japan), and Universal primer (Tokuyama dental Corp., Taitou, Taitou-ku, Tokyo, Japan) respectively. The last group was the control with no surface treatment. Dual cured resin cement (Biscem, Bisco Inc., Schaumburg, IL, USA) was luted to each group of specimens. And then, shear bond strengths were measured by universal tesing machine. The significance of the result was statistically analyzed by one-way ANOVA and Tukey test. The failure sites in each group were inspected under a magnifier. Results: Mean shear bond strength were 0.60, 1.39, 1.03, 1.38 MPa for control, Zirconia Liner (ZL), Alloy primer (AP), Universal primer (UP), respectively. Groups with application of each of the three primers showed significantly higher shear bond strength compared to the control group (p < 0.05). Among the three groups with the treatment, ZL and UP showed significantly higher shear bond strength than AP (p < 0.05), and there were no significant differences in mean shear bond strength between ZL and UP (p < 0.05). While the most specimens of control groups showed adhesive failure (80%), the most specimens of three primer-treated groups showed cohesive or mixed failure (80%).

Keywords: primer, resin cement, shear bond strength, zirconia

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Authors: Andreea Molnar, Amalia Ardeljan, Lexi Frankel, Marissa Dallara, Brittany Nagel, Omar Rashid

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Background: Escherichia coli is a gram-negative, facultative anaerobic bacteria that belongs to the family Enterobacteriaceae and resides in the intestinal tracts of individuals. E.Coli has numerous strains grouped into serogroups and serotypes based on differences in antigens in their cell walls (somatic, or “O” antigens) and flagella (“H” antigens). More than 700 serotypes of E. coli have been identified. Although most strains of E. coli are harmless, a few strains, such as E. coli O157:H7 which produces Shiga toxin, can cause intestinal infection with symptoms of severe abdominal cramps, bloody diarrhea, and vomiting. Infection with E. Coli can lead to the development of systemic inflammation as the toxin exerts its effects. Chronic inflammation is now known to contribute to cancer development in several organs, including the prostate. The purpose of this study was to evaluate the correlation between E. Coli and the incidence of prostate cancer. Methods: Data collected in this cohort study was provided by a Health Insurance Portability and Accountability Act (HIPAA) compliant national database to evaluate patients infected with E.Coli infection and prostate cancer using the International Classification of Disease (ICD-10 and ICD-9 codes). Permission to use the database was granted by Holy Cross Health, Fort Lauderdale for the purpose of academic research. Data analysis was conducted through the use of standard statistical methods. Results: Between January 2010 and December 2019, the query was analyzed and resulted in 81, 037 patients after matching in both infected and control groups, respectively. The two groups were matched by Age Range and CCI score. The incidence of prostate cancer was 2.07% and 1,680 patients in the E. Coli group compared to 5.19% and 4,206 patients in the control group. The difference was statistically significant by a p-value p<2.2x10-16 with an Odds Ratio of 0.53 and a 95% CI. Based on the specific treatment for E.Coli, the infected group vs control group were matched again with a result of 31,696 patients in each group. 827 out of 31,696 (2.60%) patients with a prior E.coli infection and treated with antibiotics were compared to 1634 out of 31,696 (5.15%) patients with no history of E.coli infection (control) and received antibiotic treatment. Both populations subsequently developed prostate carcinoma. Results remained statistically significant (p<2.2x10-16), Odds Ratio=0.55 (95% CI 0.51-0.59). Conclusion: This retrospective study shows a statistically significant correlation between E.Coli infection and a decreased incidence of prostate cancer. Further evaluation is needed in order to identify the impact of E.Coli infection and prostate cancer development.

Keywords: E. Coli, prostate cancer, protective, microbiology

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Authors: Ryan B. Simpson, Margaret A. Waskow, Aishwarya Venkat, Elena N. Naumova

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The Centers for Disease Control and Prevention (CDC) estimate that 31 known foodborne pathogens cause 9.4 million cases of these illnesses annually in US. Over 90% of these illnesses are associated with exposure to Campylobacter, Cryptosporidium, Cyclospora, Listeria, Salmonella, Shigella, Shiga-Toxin Producing E.Coli (STEC), Vibrio, and Yersinia. Contaminated products contain parasites typically causing an intestinal illness manifested by diarrhea, stomach cramping, nausea, weight loss, fatigue and may result in deaths in fragile populations. Since 1998, the National Outbreak Reporting System (NORS) has allowed for routine collection of suspected and laboratory-confirmed cases of food poisoning. While retrospective analyses have revealed common pathogen-specific seasonal patterns, little is known concerning the stability of those patterns over time and whether they can be used for preventative forecasting. The objective of this study is to construct a calendar of foodborne outbreaks of nine infections based on the peak timing of outbreak incidence in the US from 1996 to 2017. Reported cases were abstracted from FoodNet for Salmonella (135115), Campylobacter (121099), Shigella (48520), Cryptosporidium (21701), STEC (18022), Yersinia (3602), Vibrio (3000), Listeria (2543), and Cyclospora (758). Monthly counts were compiled for each agent, seasonal peak timing and peak intensity were estimated, and the stability of seasonal peaks and synchronization of infections was examined. Negative Binomial harmonic regression models with the delta-method were applied to derive confidence intervals for the peak timing for each year and overall study period estimates. Preliminary results indicate that five infections continue to lead as major causes of outbreaks, exhibiting steady upward trends with annual increases in cases ranging from 2.71% (95%CI: [2.38, 3.05]) in Campylobacter, 4.78% (95%CI: [4.14, 5.41]) in Salmonella, 7.09% (95%CI: [6.38, 7.82]) in E.Coli, 7.71% (95%CI: [6.94, 8.49]) in Cryptosporidium, and 8.67% (95%CI: [7.55, 9.80]) in Vibrio. Strong synchronization of summer outbreaks were observed, caused by Campylobacter, Vibrio, E.Coli and Salmonella, peaking at 7.57 ± 0.33, 7.84 ± 0.47, 7.85 ± 0.37, and 7.82 ± 0.14 calendar months, respectively, with the serial cross-correlation ranging 0.81-0.88 (p < 0.001). Over 21 years, Listeria and Cryptosporidium peaks (8.43 ± 0.77 and 8.52 ± 0.45 months, respectively) have a tendency to arrive 1-2 weeks earlier, while Vibrio peaks (7.8 ± 0.47) delay by 2-3 weeks. These findings will be incorporated in the forecast models to predict common paths of the spread, long-term trends, and the synchronization of outbreaks across etiological agents. The predictive modeling of foodborne outbreaks should consider long-term changes in seasonal timing, spatiotemporal trends, and sources of contamination.

Keywords: foodborne outbreak, national outbreak reporting system, predictive modeling, seasonality

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Authors: Miguel García-Ferrús, Yolanda Domínguez, M Angeles Castillo, M Antonia Ferrús, Ana Jiménez-Belenguer

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Antibiotic resistance is a growing public health concern worldwide, and it is now regarded as a critical issue within the "One Health" approach that affects human and animal health, agriculture, and environmental waste management. This concept focuses on the interconnected nature of human, animal and environmental health, and WHO highlights zoonotic diseases, food safety, and antimicrobial resistance as three particularly relevant areas for this framework. Fresh vegetables are garnering attention in the food chain due to the presence of pathogens and because they can act as a reservoir for Antibiotic Resistance Bacteria (ARB) and Antibiotic Resistance Genes (ARG). These fresh products are frequently consumed raw, thereby contributing to the spread and transmission of antibiotic resistance. Therefore, the aim of this research was to study the microbiological quality, the prevalence of ARB, and their role in the dissemination of ARG in fresh vegetables intended for human consumption. For this purpose, 102 samples of fresh vegetables (30 lettuce, 30 cabbage, 18 strawberries and 24 spinach) from different retail establishments in Valencia (Spain) have been analyzed to determine their microbiological quality and their role in spreading ARB and ARG. The samples were collected and examined according to standardized methods for total viable bacteria, coliforms, Shiga toxin-producing Escherichia coli (STEC), Listeria monocytogenes and Salmonella spp. Isolation was made in culture media supplemented with antibiotics (cefotaxime and meropenem). A total of 239 strains resistant to beta-lactam antibiotics (Third-Generation Cephalosporins and Carbapenems) were isolated. Thirty Gram-negative isolates were selected and biochemically identified or partial sequencing of 16S rDNA. Their sensitivity to 12 antibiotic discs was determined using the Kirby-Bauer disc diffusion technique to different therapeutic groups. To determine the presence of ARG, PCR assays for the direct sample and selected isolate DNA were performed for main expanded spectrum beta-lactamase (ESBL)-, carbapenemase-encoding genes and plasmid-mediated quinolone resistance genes. From the total samples, 68% (24/24 spinach, 28/30 lettuce and 17/30 cabbage) showed total viable bacteria levels over the accepted standard 10(2)-10(5) cfu/g range; and 48% (24/24 spinach, 19/30 lettuce and 6/30) showed coliforms levels over the accepted standard 10(2)-10(4) cfu/g range. In 9 samples (3/24 spinach, 3/30 lettuce, 3/30 cabbage; 9/102 (9%)) E. coli levels were higher than the standard 10(3) cfu/g limit. Listeria monocytogenes, Salmonella and STEC have not been detected. Six different bacteria species were isolated from samples. Stenotrophomonas maltophilia (64%) was the prevalent species, followed by Acinetobacter pitii (14%) and Burkholderia cepacia (7%). All the isolates were resistant to at least one tested antibiotic, including meropenem (85%) and ceftazidime (46%). Of the total isolates, 86% were multidrug-resistant and 68% were ESBL productors. Results of PCR showed the presence of resistance genes to beta-lactams blaTEM (4%) and blaCMY-2 (4%), to carbapenemes blaOXA-48 (25%), blaVIM (7%), blaIMP (21%) and blaKPC (32%), and to quinolones QnrA (7%), QnrB (11%) and QnrS (18%). Thus, fresh vegetables harboring ARB and ARG constitute a potential risk to consumers. Further studies must be done to detect ARG and how they propagate in non-medical environments.

Keywords: ESBL, β-lactams, resistances, fresh vegetables.

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Authors: Kinga Wieczorek, Elzbieta Kukier, Remigiusz Pomykala, Beata Lachtara, Renata Szewczyk, Krzysztof Kwiatek, Jacek Osek

Abstract:

The MICRORISK project (Research cooperation in assessment of microbiological hazard and risk in the food chain) was funded by the European Commission under the FP7 PEOPLE 2012 IRSES call within the International Research Staff Exchange Scheme of Marie Curie Action and realized during years from 2014 to 2015. The main aim of the project was to establish a cooperation between the European Union (EU) and the third State in the area important from the public health point of view. The following organizations have been engaged in the activity: National Veterinary Research Institute (NVRI) in Pulawy, Poland (coordinator), French Agency for Food, Environmental and Occupational Health & Safety (ANSES) in Maisons Alfort, France, National Scientific Center Institute of Experimental and Clinical Veterinary Medicine (NSC IECVM), Kharkov and State Scientific and Research Institute of Laboratory Diagnostics and Veterinary and Sanitary Expertise (SSRILDVSE) Kijev Ukraine. The results of the project showed that Ukraine used microbiological criteria in accordance with Commission Regulation (EC) No 2073/2005 of 15 November 2005 on microbiological criteria for foodstuffs. Compliance concerns both the criteria applicable at the stage of food safety (retail trade), as well as evaluation criteria and process hygiene in food production. In this case, the Ukrainian legislation also provides application of the criteria that do not have counterparts in the food law of the European Union, and are based on the provisions of Ukrainian law. Partial coherence of the Ukrainian and EU legal requirements in terms of microbiological criteria for food and feed concerns microbiological parameters such as total plate count, coliforms, coagulase-positive Staphylococcus spp., including S. aureus. Analysis of laboratory methods used for microbiological hazards control in food production chain has shown that most methods used in the EU are well-known by Ukrainian partners, and many of them are routinely applied as the only standards in the laboratory practice or simultaneously used with Ukrainian methods. The area without any legislation, where the EU regulation and analytical methods should be implemented is the area of Shiga toxin producing E. coli, including E. coli O157 and staphylococcal enterotoxin detection. During the project, the analysis of the existing Ukrainian and EU data concerning the prevalence of the most important food-borne pathogens on different stages of food production chain was performed. Particularly, prevalence of Salmonella spp., Campylobacter spp., L. monocytogenes as well as clostridia was examined. The analysis showed that poultry meat still appears to be the most important food-borne source of Campylobacter and Salmonella in the UE. On the other hand, L. monocytogenes were seldom detected above the legal safety limit (100 cfu/g) among the EU countries. Moreover, the analysis revealed the lack of comprehensive data regarding the prevalence of the most important food-borne pathogens in Ukraine. The results of the MICRORISK project are networking activities among researches originations participating in the tasks will help with a better recognition of each other regarding very important, from the public health point of view areas such as microbiological hazards in the food production chain and finally will help to improve food quality and safety for consumers.

Keywords: cooperation, European Union, food chain safety, food law, microbiological risk, Microrisk, Poland, Ukraine

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