Carlos Augusto Fernandes Oliveira

Artificial ripening of climacteric fruits remains a common postharvest practice in Pakistan and other developing countries, yet its effects on sapodilla ( Manilkara zapota ) quality and food safety remain uncharacterized. This study assessed the impact of regularly used artificial ripening methods, including ethylene gas, a commercial ethylene‐based ripener, and calcium carbide (CaC ₂ ), on physicochemical quality parameters, nutritional composition, selected antioxidant‐related indicators, and potentially toxic elements (PTEs) in sapodilla fruit pulp. Dietary exposure and non‐carcinogenic risks were evaluated using an established dietary intake model. CaC ₂ treatment significantly reduced total soluble solids, ascorbic acid, proximate nutritional composition parameters, and antioxidant indicators. In addition, CaC ₂ ‐treated fruits had levels of arsenic (0.22 mg/kg) and cadmium (0.14 mg/kg) exceeding their respective maximum levels internationally recommended. In contrast, controlled ethylene gas preserved key quality attributes of sapodilla with low PTE levels. The total hazard index (ΣTHI) for all treatments remained below the threshold (< 1.0), indicating no immediate non‐carcinogenic risk under the applied assumptions. However, carcinogenic risk estimates for arsenic (6.6 × 10 ⁻⁴ ) and cadmium (1.71 × 10 ⁻³ ) in CaC ₂ ‐treated fruits exceeded the conventional acceptable benchmark of 1 × 10 ⁻⁴ , indicating a potential long‐term carcinogenic concern under the modeled exposure assumptions. These findings highlight the need for routine monitoring of ripening agents and enforcement of food control measures in Pakistan to meet regulatory demands and protect consumer health.

Aflatoxin (AF) exposure during pregnancy may compromise maternal health and fetal development. This study aimed to evaluate the effects of gestational exposure to AF in pregnant Wistar rats and the mitigating effect of a functional yogurt (FY) containing inactivated biomass of Lacticaseibacillus rhamnosus and Saccharomyces cerevisiae. Animals were exposed to total AF (sum AFB₁, AFB₂, AFG₁ and AFG₂: 1.0 mg/kg/day) from gestational day (GD) 6 to 15, with or without FY supplementation. Maternal and reproductive parameters, fetal development, cranial ossification and placental growth factor (PlGF) immunostaining were evaluated, along with determination of AF metabolites in maternal and fetal samples. AF exposure reduced maternal weight gain during GD6–GD15 and fetal weight at GD20. Delayed cranial ossification occurred in 51.4 and 15.6% of fetuses in AF and AF + FY groups, respectively, with most severe alterations occurring in the AF group. Serum aflatoxin M₁ (AFM₁) concentrations were lower (p = 0.002) in AF + FY than in AF groups. AF metabolites were quantified less frequently in maternal liver from FY-supplemented animals (p = 0.015), while placentas or fetal livers from the AF + FY group had no quantifiable levels of AF metabolites. PlGF immunostaining was highest in AF group and lowest in the FY group (p = 0.0007). In conclusion, FY containing L. rhamnosus and S cerevisiae biomasses attenuated some AF-induced effects on fetal development, particularly delayed cranial ossification.

Mead is a fermented beverage produced through the yeast fermentation of diluted honey. Among its variations, melomel stands out as it is prepared with the addition of fruits to enhance the sensory and nutritional profile of the beverage. Although Saccharomyces cerevisiae is the conventional choice for mead fermentation, the probiotic yeast Saccharomyces cerevisiae var. boulardii has emerged as a promising starter culture to develop functional alcoholic beverages. This study aimed to evaluate the sensory profile of a cupuaçu (Theobroma grandiflorum) melomel fermented with S. boulardii using the consumers acceptance and the Check-All-That-Apply (CATA) methodologies. Accordingly, two treatments were designed for the sensory analysis: (i) Treatment 1 (T1), consisting of cupuaçu melomel fermented with the probiotic yeast S. boulardii at a concentration of 1 g/L; and (ii) Treatment 2 (T2), consisting of cupuaçu melomel fermented with the commercial yeast S. cerevisiae, also at a concentration of 1 g/L. The products were evaluated by 113 healthy volunteer consumers, aged 18 years or older, who were not pregnant and had no history of allergy or intolerance to alcohol, honey, and/or cupuaçu. Although the participants had limited familiarity with honey wine, all reported being familiar with and enjoying both honey and cupuaçu. Compared with the cupuaçu melomel fermented with the conventional yeast S. cerevisiae, the melomel fermented with S. boulardii achieved higher scores (p < 0.05) for flavor, overall impression and purchase intent. CATA analysis revealed that attributes indicating “light amber color,” “clear,” “honey aroma,” “sweet taste,” “honey flavor,” and “Flavorful” were associated with the S. boulardii formulation. In conclusion, S. boulardii represents a viable alternative culture for melomel processing, supporting the development of fermented beverages with improved sensory characteristics and good consumer acceptance.

Inactivated microorganisms may reduce mycotoxins’ bioavailability, although in vivo evidence is limited especially in farm animals. This pilot study evaluated whether a fermented milk (FM) containing heat-inactivated biomasses of Lacticaseibacillus rhamnosus and Saccharomyces cerevisiae at 1% ( w/w ) mitigated the toxic effects of dietary aflatoxins (AFs) and fumonisins (FBs) in weaned piglets. Twenty-four male crossbred piglets were assigned to a 2 × 2 × 2 factorial design ( n = 3 per group) with AFs (0; 1.11–1.28 mg/kg, sum of AFB ₁ , AFB ₂ , AFG ₁ and AFG ₂ ), FBs (0.78–0.99; 9.22–9.53 mg/kg, sum of FB ₁ and FB ₂ ), and FM (0; 39 g/animal/day, equivalent to 5% of basal diet) for 28 days. AFs reduced ( P < 0.05) the body weight gain and feed consumption. AFs or AFs + FBs exposure were associated with hepatic lesions, whereas FBs caused increased pulmonary and renal lesions ( P < 0.05). FM was associated with lower ( P < 0.05) scores of FBs’ lung lesions and AFs + FBs’ kidney lesions, but did not consistently improve growth performance or hepatic and renal biochemical parameters. Plasma AFP ₁ and urinary FB ₂ were lower ( P < 0.05) in AFs- or FBs-exposed animals receiving FM, while plasma and urinary AFM ₁ or FB ₁ were not reduced by the FM. In conclusion, FM containing inactivated L. rhamnosus and S. cerevisiae had partial protective effects against organ-specific mycotoxin injury, with limited evidence of AFs bioavailability reduction, considering n = 3 per experimental group. Further studies should evaluate different inclusion levels of microorganisms and mycotoxins, as well as other potentially adsorptive microorganisms, with a focus on microorganism–mycotoxin interactions within the gastrointestinal tract.

Mycotoxins are toxic fungal secondary metabolites that can contaminate the feed of dairy animals and can be transferred into milk and dairy products. This study aims to assess the occurrence of major mycotoxins in 190 samples of milk-based infant foods commercially available in São Paulo, Brazil, and characterize the associated risks. Aflatoxins (AFs) M₁ (AFM₁), B₁ (AFB₁), B₂ (AFB₂), G₁ (AFG₁) and G₂ (AFG₂), fumonisins (FBs) B₁ (FB₁) and B₂ (FB₂), ochratoxin A (OTA), zearalenone (ZEN), deoxynivalenol (DON), and T-2 toxin were analyzed using ultra-performance liquid chromatography coupled with tandem mass spectrometry (UPLC–MS/MS). Ninety-eight samples (51.6%) were contaminated with at least one mycotoxin, while the co-occurrence of two to four mycotoxins was observed in 32 samples (16.8%). ZEN was the most prevalent mycotoxin (60 samples; 32%), followed by FBs (FB₁ and FB₂) (42 samples; 22%), OTA (11 samples; 5.8%), DON (8 samples; 4.2%), and T-2 toxin (2 samples; 1.1%). AFs were quantified in only two samples (1.1%), one milk-based product containing 0.630 μg/kg of AFM₁ and one flavored milk powder with AFB₁ at 5.850 μg/kg. The analyzed flavored milk powder samples showed the highest maximum concentrations of DON, FBs, and OTA (165.5, 93.79, and 86.77 μg/kg, respectively), while unflavored milk powder contained the highest concentration of ZEN (17.40 μg/kg). Infant formula provided the highest estimated daily intake (EDI) value of DON (1.226 μg/kg bw/day) for infants aged ≤one year. Hazard quotient (HQ) values > 1 revealed a potential health concern for DON in only one sample of infant formula intended for infants up to six months of age. Margin of exposure (MoE) values for AFs and OTA were <10,000, emphasizing the need for continuous monitoring due to their potential toxic effects. These findings emphasize the need for stringent preventive measures and regulatory compliance to prevent mycotoxin contamination in milk-based infant foods in Brazil.

Background: Mycotoxins are toxic secondary fungal metabolites that can be transferred into milk of lactating animals that consumed mycotoxin‐contaminated feed, posing health concern for consumers. Brazil has climatic conditions that favour the growth of toxigenic fungi. Aim: This review aimed to critically examine the occurrence of mycotoxins, particularly aflatoxin M ₁ (AFM ₁ ) in Brazilian milk and dairy products in the last 20 years. Toxicological aspects of mycotoxins, regulations, preventive and potential decontamination strategies are also discussed. Methods: A comprehensive review of peer‐reviewed journal articles published in English or Portuguese from 2005 to 2025 was conducted. Sources were selected based on relevance to mycotoxin occurrence in Brazilian fluid milk and dairy products. Major Findings: The reviewed studies analysed 3939 samples of milk and dairy products, of which 2240 (56.8%) were found to contain at least one type of mycotoxin. AFM ₁ was the most frequently detected, followed by ochratoxin A, fumonisins and deoxynivalenol. Most studies reported AFM ₁ levels in milk within the Brazilian maximum limit (ML) of 0.5 μg/kg, while 11.3% of samples would exceed the European ML (0.05 μg/kg). Scientific or Industrial Implications: The occurrence levels of mycotoxins reported in Brazilian milk and dairy products represent a health threat, particularly to vulnerable populations such as children and the elderly. Continuous control and routine surveillance are essential to minimise the dietary exposure of Brazilian dairy herds to mycotoxins.

Bananas ( Musa spp.), a widely cultivated and consumed fruit, are subjected to various artificial ripening agents to meet market demands in developing countries. This study examined the effects of different ripening methods on the physicochemical, nutritional quality, and potentially toxic elements (PTEs) contamination in bananas. Health risk assessments were conducted to evaluate safety of ripening treatments. Samples treated with calcium carbide (CaC ₂ ) had the highest percentage of total soluble solids (TSS) (25.38°Brix) and TSS/titratable acidity (TA) ratio (121.06), indicating rapid ripening but with the lowest vitamin C level (5.10 mg/100 mL) and antioxidant potential. Untreated bananas retained the highest total phenolic content (TPC), antioxidant activity, and vitamin C level, highlighting their superior nutritional and bioactive properties. Ethylene gas‐treated bananas maintained balanced nutritional attributes with relatively high antioxidant activity, while ethylene ripener‐treated bananas showed intermediate results. CaC ₂ ‐treated bananas contained detectable levels of lead (0.45 mg/kg), cadmium (0.51 mg/kg), and arsenic (0.32 mg/kg), which exceeded safe limits regarding health risk indices. Untreated and ethylene‐treated bananas exhibited minimal PTE contamination and negligible health risks. Arsenic posed the highest carcinogenic risk, especially in CaC ₂ ‐ripened banana (2.4 × 10 ⁻³ ), followed by ethylene ripener (1.05 × 10 ⁻³ ). CaC ₂ ‐ripened samples also contributed to carcinogenic risk for cadmium (9.6 × 10 ⁻⁴ ). These findings provide scientific evidence to support the adoption of ethylene‐based methods as a safer alternative for ripening of bananas. In addition, the data presented stress the need for stricter regulations on artificial ripening agents for fruits, especially CaC ₂ , to safeguard public health in Pakistan and other developing countries.