Raw milk sales are legal in 30 states, including Mississippi, where regulations are largely management-based and not food-safety related. The research objectives were to analyze the microbial profile of raw goat milk from Mississippi farms and assess food safety practices and perceptions of dairy goat farmers. Deficiencies and opportunities for improvement in raw goat milk safety were identified. Staphylococcus spp. was present in most (70%) milk samples but aligned with drying-off. E. coli, Enterobacteriaceae, and coliforms were observed in 4, 10, and 12 samples, respectively; no Salmonella spp. or Listeria monocytogenes were detected. The survey highlighted farmers' need for educational opportunities on safe milk handling with emphasis on handwashing. Most producers (80.8%) are willing to invest more in their farms, creating strategic opportunities for extension outreach to help with market expansion and sustainability. Overall, training accessibility could help producers nationwide by encouraging safe milk handling and best management practices.

Bifidobacteria are the predominant members of the infant gut, colonize adults to a lesser extent, and are recognized as beneficial microbes. Various bifidobacterial species produce ��-aminobutryic acid (GABA), the chief inhibitory neurotransmitter in the mammalian central nervous system. It is postulated that in order to produce GABA, the bifidobacterial genome must contain the gadB and gadC genes which encode a glutamate decarboxylase and a glutamate/GABA antiporter, respectively. Once exported by GadC, GABA is absorbed and transported systemically throughout the host. We hypothesize that specific dietary oligosaccharides will modulate bifidobacterial production of GABA due to varying intracellular concentrations of glutamate. To test this, 33 bifidobacterial strains were screened for GABA production via reverse phase HPLC. Interestingly, 10 strains contained both gadB and gadC genes, but only 8 strains produced detectable GABA in vitro. To further elucidate the extrinsic factors influencing GABA production, strains were subjected to different dietary components. Specifically, lactose and the dietary oligosaccharide FOS were evaluated for the ability to promote biosynthesis of intracellular glutamate and thus potentially GABA. Understanding the relationship between diet, bifidobacterial physiology, and GABA production may inform dietary interventions to modulate this neurotransmitter in vivo.

Antimicrobial coatings in which the active agent (e.g. N-halamine) can regenerate activity represent a promising way to prevent microbial cross-contamination. A reported method for applying coatings containing antimicrobial N-halamines is layer-by-layer (LbL) application of polyelectrolytes, which form N-halamines upon cross-linking. Prior reports on dip layer-by-layer (LbL) fabrication have demonstrated the potential of this coating technology; however, spray LbL fabrication would enable more rapid coating and represents a more commercially translatable application technique. In this work, dip and spray LbL methods were used to coat polypropylene (PP) with N-halamine containing bilayers consisting of cross-linked polyethylenimine (PEI) and poly(acrylic acid) (PAA). Further experimentation with spray LbL fabrication used naturally occurring polyelectrolytes, chitosan and alginate. Materials were characterized using atomic force microscopy (AFM), ellipsometry, contact angle, fourier transform infrared spectroscopy, a chlorine content assay, and a dye assay for amine quantification. All methods of coating application exhibited a 99.999% (5-log) reduction against Listeria monocytogenes with application time for spray LbL taking less than 10% of the time required for dip LbL. Spray LbL fabrication of N-halamines is a rapid and inexpensive method to fabricate rechargeable antimicrobial surfaces.

The iron chelating molecule, ethylenediaminetetraacetic acid (EDTA) is used in food applications for the preservation of oxidation prone ingredients. Research has suggested that EDTA is also capable of enhancing the antimicrobial effectiveness of various compounds including naturally-derived antimicrobials. With consumer demand for cleaner food labels, there remains an opportunity to introduce new chelating technology to replace synthetically-derived EDTA. Through photographting and chemical conversion, hydroxamic acid ligands were covalently bound to polypropylene films resulting in polypropylene-graft-poly(hydroxamic acid) (PP-g-PHA). The resulting films demonstrated an ability to chelate 64 nmol/cm2 from an iron saturated environment or 163 nmol/cm2 of magnesium and 139 nmol/cm2 of calcium from bacterial growth media. A surface pKa of 8.97 suggested that film ligands should remain protonated under acidic and neutral pH conditions. When combined with lysozyme, PP-g-PHA films were able to reduce inhibitory concentration of lysozyme for Listeria monocytogenes by half. When tested against Bacillus cereus, Pseudomonas fluorescens, and E. coli O157:H7; PP-g-PHA films were unable to inhibit growth and showed little enhancement of lysozyme. EDTA controls revealed that similar levels of soluble chelator were more effective than immobilized chelators. EDTA results also suggested that a chelating film with a higher affinity for iron (through coordination or ligand stability) may be able to control B. cereus growth. Both EDTA and PP-g-PHA caused P. fluorescens to produce siderophores (pyoerdines), suggesting that each treatment resulted in a low-iron growth environment. These findings suggest that surface bound chelating technology can affect the growth of L. monocytogenes and enhance the effectiveness of lysozyme. With improved surface chemistry (a higher binding constant with iron), this technology has the potential to influence the growth of other pathogens and spoilage microorganisms.

The purpose of this investigation is to determine through routine checks the number of organisms present in the various raw ingredients used for the preparation of foods. The problem has consisted of, first, a determination of the total bacterial population by numbers; second, a determination of the incidence of colon bacteria in the samples examined; third, a determination of the presence of yeasts and molds; and fourth, an attempt to utilize this information obtained in judging the methods of handling the raw ingredients before they are used in the processing of foods.

The purposes of this study are, first, to determine relative numbers and predominant types of bacterial organisms in the various food products studied; second, to evaluate the efficiency of the paper coverings of these foods; and third, to determine whether these products are potential sources of food poisoning.

Thesis (M. Tech. Environmental health) -- Central University of technology, Free State, 2011 / Although the application of low pH is common practice in food preservation, the emergence of acid tolerance has been reported world-wide amidst a growing concern that preservation with weak acids, such as organic acids may be influenced as a result of food-borne bacteria becoming acid tolerant or acid resistant. The present study was conducted to assess the acid tolerance of a wide range of bacterial species and consequently the sustainable application of organic acids as food preservatives in particularly acidic foodstuffs. Acid tolerance was determined in 19 bacterial strains predominantly associated with food spoilage and food poisoning. After exposure to hydrochloric acid 16% of the isolates were found to be intrinsically tolerant to low pH and included amongst others the enteric bacteria Escherichia coli and Salmonella spp. The latter organisms are known causative agents in food spoilage and poisoning, and the results highlight the predicaments related to their ability to survive in acidic foodstuffs as well as the human gastric environment. Bacterial strains were further exposed to increasing concentrations of various acidic foodstuffs in order to determine the development of acid tolerance by gradual decrease in pH, as opposed to exposure to acid shock. After induction, the protein profiles of resulting acid tolerant isolates were compared with those of the original un-induced strains. Exposure to acidic foodstuffs resulted in various survival profiles, where not only pH but also the type of acidulant (foodstuff or inorganic acid) were found to be contributing factors in acid tolerance development. Bacterial protein composition after exposure to acidic foodstuffs showed considerable variation which may be indicative of acid tolerance development whereas the mechanisms involved may be the result of multiple modifications in bacterial composition. After the induction of acid tolerance, susceptibility of induced strains to various organic acids were determined at various pH values. This was done to investigate whether acid tolerance would influence the inhibitory activity of organic acids as antimicrobial agents in acidic food. Decreased susceptibility was not significantly demonstrated with the exception of only selected isolates, the latter including E. coli and S. typhimurium. Organic acid activity was found to be much more effective at lower pH values and it would be necessary to elucidate whether this inhibition is the result of a lower pH or more specifically the activity of the organic acids. The effect of exposure to an acidic environment on phenotypic characteristics of Gram-negative bacteria, and more specifically psychrotrophic organisms was evaluated in order to show the combined effect of organic acids and low temperature preservation. The characteristic yellow pigment of various Chryseobacterium species was found to be not as apparent after acid exposure while in some cases the colonies were observed as white. In Pseudomonas aeruginosa the characteristic green pigment was much more prominent after acid exposure. These morphological alterations may be important factors that should be considered in identification procedures employed in food safety laboratories. Finally, the influence of acidic exposure via acidic foodstuffs and also organic acids on the protein composition and outer membrane protein structure of various bacterial cells was investigated. No specific relationships with the MICs (Minimum Inhibitory Concentrations) of organic acids after induction with the selected acidic foodstuffs could be established, although various differences were found in protein expression. From the results, it may be suggested that the outer membrane of various pathogenic bacteria is involved in acid tolerance development and this supports the reports on the importance of membrane integrity in the protection against low pH. In conclusion, the study endeavoured to add to the body of knowledge with regard to alternative food preservation regimes utilising organic acids, either solely or in combination with selected extrinsic and intrinsic parameters.

Farmers’ markets have increasingly become a popular venue for purchase of fresh, locally-grown produce, with the number of farmers’ markets in Kentucky reaching an all-time high of 159 in 2016. Good Agriculture Practices (GAPs) is a program created by the USDA’s Agriculture Marketing Service to function as a food safety audit for small-scale fresh produce growers, such as those who sell fresh produce at local farmers’ markets. However, under the provisions of the Food Safety Modernization Act of 2011, small-scale farmers who sell an average of $25,000 in annual fresh produce sales across the span of three years are exempt from mandatory food safety certification. Many smallscale farmers in Kentucky fall below this threshold, and do not hold food safety certification. This study had two objectives: to investigate the practices, perceptions, and implementation of GAPs among small-scale Kentucky farmers who sell at farmers’ markets; and to create and evaluate the effectiveness of commodity-specific informational factsheets to disseminate food safety knowledge among small-scale Kentucky farmers. Data from the perceptions, practices, and implementation survey were analyzed from 160 completed surveys of small-scale fresh produce growers on-site at farmers’ markets in 21 counties across the state of Kentucky (see Appendix A). The results were mixed, with 90% of participants indicated familiarity with GAPs, but only 47% opting to practice water quality GAPs and 55% choosing to observe soil amendment GAPs. Participants did report slightly higher compliance with field sanitation (71%) and sanitary facilities (73%) GAPs, but indicated that cost (67%) and time (68%) were significant perceived barriers to completing a GAPs audit on their farm. Participants also failed to identify many sources of potential microbiological contamination, with soil only being identified as a source of pathogenic contamination by 41% of participants and irrigation water identified by 51% of participants. Even fewer participants believed that contamination could result from ice (26%) or refrigeration and cooling (28%). However, most respondents indicated a desire to undergo further GAPs education, and the factsheet evaluation data indicated that the factsheets were highly effective and had resulted in significant GAPs knowledge increases for participants.

Amostras de açúcar mascavo disponíveis em diferentes estabelecimentos comerciais no município de Araras - SP foram adquiridas para análises. No total foram coletados 49 pacotes de açúcar durante um período de quatro meses. As análises foram realizadas com o objetivo de avaliar a qualidade desse produto oferecido ao consumidor. As análises microbiológicas foram realizadas considerando a pesquisa dos seguintes microrganismos: bactérias mesófilas, bolores e leveduras, esporos de bactérias termófilas flat-sour, esporos de bactérias termófilas anaeróbias produtoras de H2S, esporos de bactérias termófilas anaeróbias não produtoras de H2S, Salmonella, coliformes totais e termotolerantes. Ainda foram determinadas a umidade (%) e atividade de água das amostras. Os resultados das análises apresentaram variações nas concentrações dos microrganismos estudados, cabendo às bactérias mesófilas e aos esporos de termófilas flat-sour as contagens mais elevadas, seguidos por bolores e leveduras. Todas as marcas apresentaram ausência de Salmonella, coliformes totais e termotolerantes, atendendo a legislação brasileira. Em relação ao padrão da National Canners Association apenas as marcas A, G, H e J estavam adequadas a essas exigências, as demais estavam em desacordo. Para o padrão da International Commission for Uniform Methods of Sugar Analysis (ICUMSA), nenhuma marca obteve resultado satisfatório em relação à qualidade microbiológica. Os teores de umidade variaram de 1,94 a 3,63% e os valores de atividade de água variaram de 0,55 a 0,64, havendo diferença significativa ao nível de 5% de significância. Pelos resultados obtidos considera-se que as marcas comercializadas estão de acordo com a legislação vigente no país, porém a maioria das marcas não atende aos padrões internacionais. / Samples of brown sugar available in different establishments in the city of Araras - SP were acquired for analysis. In the total 49 packages of sugar were collected during a period of four months. The analyses were performed to evaluate the quality of that product for the consumer. Microbiological tests were performed to search the following microorganisms: mesophilic bacteria, yeasts and molds, thermophilic flat-sour spores, thermophilic anaerobic H2S-producing spores, thermophilic anaerobic non-producing H2S spores, Salmonella and total and thermotolerant coliforms. Also humidity (%) and water activity were determined for the samples. The analysis results presented variations in the concentrations of microorganisms studied, being the mesophilic bacteria and thermophilic \"flat-sour\" spores the ones with highest scores, followed by yeasts and molds. All brands showed absence of Salmonella and total and thermotolerant coliforms, in accordance with the Brazilian legislation. Regarding the National Canners Association standards, only brands A, G, H and J were suited to these demands, the others being in disagreement. For the International Commission for Uniform Methods of Sugar Analysis (ICUMSA), standards no brands obtained satisfactory results in relation to microbiological quality. The moisture content ranged from 1.94 to 3.63% and the values of water activity ranged from 0.55 to 0.64, significant difference at 5% significance level. By the results it is considered that the brands are marketed under the law of the country, but most brands do not meet international standards. Humidity rates ranged from 1.94 to 3.63% and water activity ranged from 0.55 to 0.64, with significant difference at 5% significance level. By the results obtained it is considered that all brands sold are in accordance to the laws of the country, but most brands do not meet the international standards.

Além das características tecnológicas das bactérias lácticas para o processamento de alimentos fermentados, novas propriedades são requeridas, como a produção de bacteriocinas. Vários desses compostos protéicos possuem atividade antimicrobiana direcionada à patógenos e apresentam potencial de uso como bioconservadores naturais de alimentos. O objetivo desse trabalho foi avaliar a viabilidade de aplicação de cinco culturas lácticas produtoras de bacteriocinas como bioconservantes em alimentos. Avaliaram-se as propriedades de segurança e probióticas dessas culturas e determinaram-se os parâmetros de produção e purificação parcial das bacteriocinas, bem como os efeitos de fatores limitantes de crescimento (cloreto de sódio e nitrito). Os resultados demonstraram que essas linhagens não possuem atividade de enzimas indicadoras de patogenicidade (termonuclease, -hemolisinas e gelatinases). A tolerância às condições ácidas variou dependendo da linhagem. Lactobacillus plantarum CTC 368 e Enterococcus avium CTC 469 apresentaram maior tolerância a valores de pH 2,0 e 3,0. Lactococcus lactis subsp. cremoris CTC 204 e Enterococcus avium CTC 483 também apresentaram 100% de sobrevivência a pH 3,0 e, mesmo com uma diminuição da tolerância a pH 2,0, a sobrevivência foi cerca de 80%. Com relação à susceptibilidade aos sais de bile, observou-se que a viabilidade variou entre as linhagens em função da concentração e tempo de incubação. Enterococcus avium CTC 469 e Lactococcus lactis subsp. hordinae CTC 484 foram as mais tolerantes e apresentaram sobrevivência de 57% e 58%, respectivamente. As linhagens mostraram excelente sobrevivência tanto a baixos valores de pH, quanto às concentrações de sais biliares indicando potencial probiótico. As características de crescimento e de produção de bacteriocinas foram semelhantes para todas as linhagens e meios avaliados. A viabilidade aumentou durante o período de incubação e a produção de bacteriocinas mostrou uma cinética de metabólito primário. A suplementação do meio MRS com 1,0% de NaCl e 0,1% de NaNO2, não afetou o crescimento de Lc. lactis subsp. cremoris CTC 204, mas promoveu a redução da atividade da bacteriocina de 42,5% e 17,5%, respectivamente. O crescimento de Lactococcus lactis subsp. hordinae CTC 484 foi afetado com a adição de NaCl e NaNO2 ao meio APT, mas ainda reteve 50% da atividade bacteriocigênica. O espectro de atividade das bacteriocinas variou de acordo com a susceptibilidade da linhagem indicadora, a concentração ou atividade da bacteriocina, a viabilidade e/ou presença das células produtoras e os procedimentos de extração das bacteriocinas. Os resultados revelaram que as bacteriocinas de Lactococcus lactis subsp. cremoris CTC 204 e Lactococcus. lactis subsp. hordinae CTC 484 promoveram um efeito bactericida contra Staphylococcus aureus. Concluiu-se que essas duas culturas produtoras de bacteriocinas apresentam propriedades para utilização como bioconservantes em alimentos e que estudos mais aprofundados, relacionados com a purificação da bacteriocina e avaliação de seu uso em alimentos, devem ser realizados. / In addition to the technological characteristics of lactic bacteria for the processing of fermented foods, other properties need to be developed and explored, such as bacteriocin production. Several of these protein-based compounds have antimicrobial activities targeted towards pathogens and have a significant potential for use as natural biopreservatives in processed foods. The objective of this study was to evaluate the viability of using five bacteriocin-producing lactic cultures as biopreservatives in foods. The safety and probiotic properties of these cultures were evaluated, followed by the determination of bacteriocin production and purification parameters and the effect of growth-limiting factors (sodium chloride and nitrite). The results demonstrated that these strains do not show activity of marker enzymes of pathogenicity (thermonuclease, - hemolysins and gelatinases). Tolerance to acidic conditions varied depending on the specific strain. Lactobacillus plantarum CTC 368 and Enterococcus avium CTC 469 exhibited greater tolerance to pH values 2,0 and 3,0. Lactococcus lactis subsp. cremoris CTC 204 and En. avium CTC 483 also presented 100% survival at pH 3,0, and even with reduced tolerance at pH 2,0, the survival rate was 80%. Variation in susceptibility to bile salts was observed between the strains as a function of concentration and incubation time. En. avium CTC 469 and Lactococcus lactis subsp. hordinae CTC 484 were found to be the most tolerant, exhibiting survival rates of 57% and 58%, respectively. The strains demonstrated excellent survival not only at low pH values, but also at high bile salt concentrations, thereby indicating probiotic potential. The growth and bacteriocin-producing characteristics were similar for all the strains and culture media evaluated. Bacterial multiplication increased during the incubation period, while bacteriocin production displayed primary metabolite kinetics. The activity spectrum of the bacteriocins varied with the susceptibility of the indicator strain, the concentration or activity of the bacteriocin, the viability and/or presence of bacteriocinproducing cells and the bacteriocin extraction procedures. The results reveal that the bacteriocins produced by Lc. lactis subsp. cremoris CTC 204 and Lc. lactis subsp. hordinae CTC 484 exerted a bactericidal effect against Staphylococcus aureus. Supplementation of the MRS medium with 1,0% NaCl and 0,1% NaNO2, did not affect the growth of Lc. lactis subsp. cremoris CTC 204, but did reduce by 42,5% and 17,5% the activity of the bacteriocin, respectively. Growth of Lc. lactis subsp. hordinae CTC 484 was affected by the addition of NaCl and NaNO2 to the APT medium, but even so still exerted 50% of its bacteriocinogenic activity. It was concluded that Lc. lactis subsp. cremoris CTC 204 and Lc. lactis subsp. hordinae CTC 484 exhibited adequate properties for use as biopreservatives in foods and that more extensive studies related to the purification of bacteriocins and assessment of their use in foods should be conducted.