Stael Málaga Carrilho

Members of the family Enterobacteriaceae are often linked to foodborne outbreaks, including acute diarrhea, with diarrheagenic Escherichia coli (DEC) as the most common cause of this disease in low‐ and middle‐income countries. A cross‐sectional study was carried out to evaluate the sanitary conditions of ready‐to‐consume milk, involving 750 pasteurized milk samples. These samples were analyzed for the Enterobacteriaceae count, the presence of Salmonella spp., and the enumeration of E. coli , as well as checking for the presence of DEC‐specific virulence genes. Two molecular assays were used to detect DEC‐specific virulence genes ( eae, bfpA, aggR, ipaH, est, elt, stx1, and stx2 ). Overall, a significant noncompliance rate (5%) was identified in the Enterobacteriaceae counts, indicating postpasteurization contamination. Two genes ( ipaH and elt ) were detected in 14.28% of E. coli ‐positive samples, highlighting the need for improved methods to minimize postpasteurization contamination in dairy plants. This improvement could contribute to better food safety standards internationally. Additionally, further studies are necessary to understand the actual risk posed by these strains circulating in milk for immunocompromised individuals or those with immature immune systems. This underscores a global concern for vulnerable populations worldwide.

Goat milk presents interesting characteristics to consumers, but the production of this food faces several challenges that influence its quality. Process failures from milking to processing and commercialization can expose milk to contamination by pathogenic microorganisms, including the coagulase-positive Staphylococcus group. Although Staphylococcus aureus is the most relevant species in mastitis and food poisoning, other species in this group are also important, especially those related to food poisoning. This study aimed to identify the coagulase-positive Staphylococcus species in goat milk using biochemical tests, determine the prevalence of antimicrobial resistance using the disc diffusion test, and investigate enterotoxin-producing genes, sea, seb, sec, sed, see, seg, seh, and sei by multiplex PCR. A total of 384 coagulase-positive Staphylococcus strains obtained from raw goat milk collected from nine farms in Paraná during four seasons of the year were studied. Biochemical tests showed that 85.69% of the 384 strains were S. aureus, followed by 9.38% of S. intermedius, 4.17% of S. hyicus and 0.78% of S. delphini. For the antimicrobial resistance test, up to three strains of each species identified as coagulase-positive Staphylococci were chosen from each farm, with a total of 74 strains. Of these, 27% (20/74) showed resistance to at least one antibiotic, and among all penicillin-resistant strains, 45% (9/20) also presented resistance to tetracycline. In the search for staphylococcal enterotoxin-producing genes, 49 S. aureus strains were studied; among them, 40.81% (20/49) presented enterotoxin-producing genes. The highest prevalence was detected for the sec gene, which was present in 22.44% (11/49) of the strains, followed by the seh gene in 18.36% (9/49), and the see gene was detected in 4.08% (2/49). It was concluded that S. aureus was the predominant species in raw goat milk, with a high prevalence of penicillin- and tetracycline-resistant Staphylococci and a significant number of strains with staphylococcal enterotoxin-producing genes. The strains studied carried enterotoxin-producing genes involved in food poisoning in humans, indicating that goat milk is a possible source of contamination and, therefore, a potential danger to public health.

Lactose is the main carbohydrate in milk, and its absorption occurs via enzymatic hydrolysis, generating glucose and galactose. Lactose intolerance is the reduction of intestinal hydrolysis capacity due to hypolactasia, which results in the need to consume dairy foods with low levels of this carbohydrate. β-galactosidase enzymes are used in dairy industries to hydrolyze lactose, thereby allowing intolerant consumers access to dairy products without the negative health implications. Alternative and official analytical methods are used to quantify the carbohydrate content resulting from enzymatic hydrolysis. The objective of this study was to evaluate the enzymatic hydrolysis of two distinct industrial enzymes produced by the microorganisms Bacillus licheniformis and Kluyveromyces lactis using three analytical methods: enzymatic method, cryoscopy, and high performance liquid chromatography (HPLC) using artificial intelligence to improve the control of the industrial processes. After adding the enzymes to skim milk, time kinetics was performed by collecting samples at time 0, every 10 min for 1 h, and every 30 min until the end of 5 h of hydrolysis. In 97% of the cases, a decrease in lactose concentration was observed by HPLC, followed by the deepening of the cryoscopic point. Glucose measurements by absorbance and HPLC quantification were correlated (r = 0.79; p < 0.01) but not concordant (p < 0.01). It was concluded that by means of artificial intelligence, it was possible to indirectly estimate lactose concentration using an algorithm that associates cryoscopy and glucose concentration.