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Pet food is increasingly recognized as a significant vehicle for the transmission of foodborne pathogens to humans. The intimate association between pets and their owners, coupled with the rising trend of feeding pets raw and unprocessed foods, contributes substantially to this issue. Salmonella contamination in pet food can originate from raw materials and feed ingredients, the processing environment, and postprocessing handling and applications. The absence of standardized postprocessing pathogen mitigation steps in the production of dry kibble and treats, along with the lack of validated heat and chemical interventions in raw pet foods, renders pet food susceptible to contamination by pathogens such as Salmonella, Listeria, E. coli, etc. Pets can then serve as carriers of Salmonella, facilitating its transmission to pet owners. Since 1999, there have been over 117 recalls of pet foods due to Salmonella contamination in the United States, with 11 of these recalls linked to human outbreaks. Notably, 5 of the 11 human outbreaks involved multidrug-resistant Salmonella strains. Various antimicrobial interventions, including high-pressure processing, ozone, irradiation, chemical treatments such as organic acids and acidulants, plant-derived antimicrobials, and biological interventions such as bacteriophages, have proven effective against Salmonella in pet foods. This review aims to summarize the prevalence of Salmonella in different types of pet foods, identify common sources of contamination, outline reported outbreaks, and discuss control measures and the regulatory framework governing pet food safety., (© 2024 The Author(s). Comprehensive Reviews in Food Science and Food Safety published by Wiley Periodicals LLC on behalf of Institute of Food Technologists.)

Mycotoxins, such as aflatoxin B 1 (AFB 1 ), deoxynivalenol (DON), fumonisins (FBs), ochratoxin A (OTA), T-2 toxin (T-2), and zearalenone (ZEN), can contaminate animal feeds and pose risks to animal health and production performance. These mycotoxins are commonly found in cereals and grains, with the increased use of cereals in pet food, there is a rising concern about mycotoxin contamination among pet owners. To address this, we analyzed imported brands of feline and canine food from the Chinese market produced in 2021-2022. Ninety-three samples were analyzed, comprising 45 feline food and 48 canine food samples. Among them, 14 were canned food and 79 were dry food. The results indicate that AFB 1 , DON, FBs, OTA, T-2, and ZEN occurred in 32.26%, 98.92%, 22.58%, 73.12%, 55.91%, and 7.53% of the samples, respectively. The most prevalent mycotoxin was DON, followed by OTA, T-2, AFB 1, and FBs, whereas ZEN was less frequently detected. The mean concentrations of the six mycotoxins in pet feed samples were 3.17 μg/kg for AFB 1 , 0.65 mg/kg for DON, 2.15 mg/kg for FBs, 6.27 μg/kg for OTA, 20.00 μg/kg for T-2, and 30.00 μg/kg for ZEN. The levels of mycotoxins were generally below the limits of the Pet Feed Hygiene Regulations of China and the EU. Notably, a substantial majority of the pet food samples (88 out of 93) were contaminated by two or more mycotoxins. AFB 1 , FBs, OTA, and ZEN occurred slightly more often in feline food than in canine food. Except for OTA, the contamination rates for the other five mycotoxins in canned food were lower than those in dry food. Moreover, except for AFB 1 , the levels of the other five mycotoxins in canned foods were lower than those in dry foods. This study highlights the widespread contamination of pet foods with mycotoxins, which poses a significant risk to pets from continuous exposure to multiple mycotoxins., (© 2024. The Author(s) under exclusive licence to Society for Mycotoxin (Research Gesellschaft für Mykotoxinforschung e.V.) and Springer-Verlag GmbH Germany, part of Springer Nature.)

The extensive utilization of pesticides in agriculture has resulted in the presence of pesticide residues in food and feed, which poses a significant threat to human health. Various physical and chemical methods have been proposed to remove pesticides, but most of these methods are either costly or susceptible to secondary contamination. Consequently, the utilization of microorganisms, such as probiotics, for eliminating pesticides, has emerged as a promising alternative. Probiotics, including lactic acid bacteria, yeasts, and fungi, have demonstrated remarkable efficiency and convenience in eliminating pesticide residues from food or feed. To promote the application of probiotic decontamination, this review examines the current research status on the utilization of probiotics for pesticide reduction. The mechanisms involved in microbial decontamination are discussed, along with the toxicity and potential health risks of degradation products. Furthermore, the review explores strategies to enhance probiotic detoxification and outlines prospects for future development.

Determining the effects of fermentation duration on the microbial ecosystem, potential pathogenic risks, and metabolite generation during the fermentation of distilled grains is essential for safeguarding the safety and enhancing the nutritional profile of animal feed. This study investigates the effect of varying fermentation times (9, 30, and 60 days) on microbial diversity, pathogenic risk, and metabolite profiles in distiller grains using 16S rDNA sequencing and LC-MS-based metabolomics. The results showed that early fermentation (9-30 days) enhanced the abundance of beneficial bacteria, such as Lactobacillus reuteri and Lactobacillus pontis ( p < 0.05), while pathogenic bacteria, like Serratia marcescens and Citrobacter freundii , were significantly reduced ( p < 0.05). Metabolomic analysis revealed an increase in unsaturated fatty acids and the degradation of biogenic amines during early fermentation. However, prolonged fermentation (60 days) led to a resurgence of pathogenic bacteria and reduced the synthesis of essential metabolites. These findings suggest that fermentation duration must be optimized to balance microbial safety and nutrient quality, with 30 days being the optimal period to reduce pathogenic risks and enhance feed quality.

Feed-based bivalent vaccine (FBBV) containing killed whole organism (KWO) of Streptococcus agalactiae and Aeromonas hydrophila with 10% palm oil was previously proved to improve red hybrid tilapia's (Oreochromis sp.) immunity against streptococcosis and Aeromonas infections. This study characterized the FBBV's stability following the preparatory process and storage. The FBBV was prepared, and the KWO's stability was determined microscopically and molecularly. The efficacy of FBBV stored at room temperature (25 ± 2 °C) for 0, 30 and 60 days was investigated in red hybrid tilapia. The results indicated the addition of palm oil was not affecting the KWO's structure and helping in the FBBV's pelletization. In 1 g of FBBV contained 1.5 × 10 9 CFU/g of S. agalactiae and 4.9 × 10 9 CFU/g of A. hydrophila, respectively, even after 60 days of storage at room temperature. The KWO's structure in FBBV was not affected following in vitro acidic tolerance analysis, as noted from light and electron microscopies. The FBBV's carbohydrate, energy, moisture, total protein and total ash contents remained stable at 95% after 60 days of storage at room temperature, while the KWO's concentration was slightly reduced to 83.3% for S. agalactiae (1.25 × 10 9 CFU/g) and 80.6% for A. hydrophila (3.85 × 10 9 CFU/g), respectively. Fish vaccinated with FBBV that was stored for 0, 30 and 60 days did not show any significant differences (p ≥ 0.05) in the relative percent survival when challenged with pathogenic Streptococcus spp. and Aeromonas spp. These findings suggested that the FBBV is a stable vaccine, which underscores its potential application as aquatic vaccines in aquaculture., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Background: With the rapid development of animal husbandry, the demand for protein feed resources is increasing. Cottonseed meal (CSM) and soybean meal (SBM) are rich sources of protein. However, their application is limited due to the existence of anti-nutrients, which can be harmful to the digestion and absorption. A strain of Bacillus subtilis (Mafic-Y7) was isolated from digesta of intestines of Tibetan pigs. The strain showed high protease activity, which helps in degrading proteinic anti-nutritional factors in grain meal and in vitro degradation of free gossypol. In order to better understand this isolated strain, whole genome of Mafic-Y7 strain was sequenced and analyzed. Different effects on various grain meals were identified., Result: The GC-depth Poisson distributions showed no bias suggesting high-quality genome assembly of Mafic-Y7. The whole genome sequencing showed that one chromosome with 4,248,845 base pairs(bp)and the genes total length with 3,736,524 bp was predicted in Mafic-Y7. Additionally, Mafic-Y7 possessed 4,254 protein-coding genes, and several protease genes were annotated by aligning them with databases. There are 55 protease genes, one phytase gene and one laccase gene were annotated in the gene sequence of Mafic-Y7. The average nucleotide identity between Mafic-Y7 and the GCA-000009045.1 homologous genome was 0.9938, suggesting a close genetic relationship between them at the species level. Compared with the closest four whole genomes, Mafic-Y7 was annotated the most abundant of protease genes (55 genes). The fermentation supernatant of Mafic-Y7 could increase the content of small peptides, water-soluble proteins, and acid-soluble proteins in vitro by 411%, 281% and 317% in SBM and 420%, 257% and 338% in CSM. After fermentation in grain meal by Mafic-Y7, the degradation rate of anti-nutritional factors in SBM, such as trypsin inhibitor, glycinin, and β-conglycinin was greater than 70%, and lectin was greater than 30%. The degradation rates of anti-nutritional factors in CSM, such as gossypol and phytic acid, were 82% and 26%, respectively., (© 2024. The Author(s).)

In this Special Issue, several interesting research and review articles were published with the aim of filling in some of the existing knowledge gaps in the field of mycotoxins [...].

Microbial communities associated with dairy farm operations have a significant influence on food safety, dairy product quality, and animal health. This study aimed to create a microbial mapping at a dairy farm to learn about their bacterial diversity, distribution, and potential dissemination pathways. The investigation included the detection of key zoonotic pathogens, enumeration of Staphylococcus aureus and Escherichia coli as indicators of typical bacterial loads in a dairy production environment, and a microbiome analysis using metagenomics. A total of 160 samples (environmental, udder swabs, feed, feces, raw milk, and water) were collected during winter (N = 80) and spring (N = 80). In winter, Cronobacter spp. were detected in four feed and two water samples; L. monocytogenes was identified in two samples, one from feces and one from a cattle mat; E. coli O157:H7 was found in two feed samples. On the other hand, during spring, Cronobacter spp. were present in four feed samples and one hallway drain, with only one feed sample testing positive for E. coli O157:H7, while L. monocytogenes was absent during the spring season. Regarding microbial counts, there was no significant difference between the two seasons (p = 0.068) for S. aureus; however, a significant difference (p = 0.025) was observed for E. coli. Environmental microbiome analysis showed the presence of Proteobacteria (46.0 %) and Firmicutes (27.2 %) as the dominant phyla during both seasons. Moraxellaceae (11.8 %) and Pseudomonadaceae (10.62 %) were notable during winter, while Lactobacillaceae (13.0 %) and Enterobacteriaceae (12.6 %) were prominent during spring. These findings offer valuable insights into microbial distribution within a dairy farm and potential risks to animal and human health through environmental cross-contamination., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Aflatoxin constitutes a significant concern for food and feed safety, posing detrimental health risks to both animals and humans. This study aimed to examine the prevalence and concentration of aflatoxins in maize feed, total mixed ration, and wheat bran collected from specialized dairy farms and local markets in three major urban centers in eastern Ethiopia. A total of 180 feed samples were collected from September 2021 to January 2022 in Chiro town, Dire Dawa city, and Harar city. These samples underwent thorough extraction and immunoaffinity clean-up before aflatoxin analysis using HPLC/FLD. The results revealed that AFB 1 , AFB 2 , AFG 1 , AFG 2 , and TAF contamination was detected in 72.2%, 66.1%, 71.1%, 68.7%, and 82.8% of the feed samples, respectively. The corresponding mean levels of each aflatoxin were 28.15 ± 3.50, 3.3 ± 0.40, 19.87 ± 1.87, 2.7 ± 0.32, and 54.01 ± 4.72 µg/kg, respectively. The occurrence and levels of aflatoxin varied across different study sites and feed types. Notably, feeds from Dire Dawa city exhibited significantly higher mean levels of AFB 1 (43.98 ± 5.3 µg/kg), AFB 2 (5.69 ± 0.6 µg/kg), AFG 1 (32.25 ± 2.7 µg/kg), and AFG 2 (5.01 ± 0.5 µg/kg) than feeds from other urban centers did. Additionally, a significantly higher occurrence of AFB 1 (29.4%) and AFG 1 (28.3%) was detected in feed from Dire Dawa city. Similarly, the total mixed ration (TMR) displayed significantly higher levels of AFB 1 (50.67 ± 5.2 µg/kg), AFB 2 (4.74 ± 0.6 µg/kg), AFG 1 (32.87 ± 2.6 µg/kg), and AFG 2 (3.86 ± 0.5 µg/kg) compared to the other feed types. Moreover, a significantly higher occurrence of AFB 1 (30.7%) and AFG 1 (28.7%) was detected in the TMR. Furthermore, a moderate correlation was observed between the count of aflatoxigenic Aspergillus species and the levels of TAF in the feed samples. Overall, this study underscores the widespread presence of aflatoxin contamination in dairy feeds in eastern Ethiopia, highlighting the urgent need for stringent monitoring and mitigation measures to ensure food and feed safety, as well as public health.

Secondary metabolic products of molds, called mycotoxins, negatively affect animal health and production. They constitute a significant problem in veterinary and medical sciences, and their presence has been confirmed in feed all over the world. Applying appropriate agricultural practices and ensuring proper storage conditions significantly reduces the contamination of agricultural products with mycotoxins. However, this does not guarantee that raw materials are completely free from contamination. Many detoxification methods are currently used, but their insufficient effectiveness and negative impact on the quality of the raw material subjected to them significantly limits their usefulness. The positive results of eliminating mycotoxins from many products have been proven by the specific properties of microorganisms (bacteria, yeast, and fungi) and the enzymes they produce. Biological detoxification methods seem to offer the most promising opportunities to solve the problem of the presence of mycotoxins in animal food. This work, based on literature data, presents the health risks to farm animals consuming mycotoxins with feed and discusses the biological methods of their purification.

In feed, propionic acid is the weak organic acid of choice to prevent growth of spoilage fungi. For safe and easy industrial handling this antifungal agent is applied in the presence of neutralizing ammonium, which however has the disadvantage to negatively affect the efficacy of fungus-inhibiting properties of the formulation. In the present study we investigated the impact of medium chain fatty acids (MCFA) on the antifungal efficacy of an ammonium propionate formulation on dormant- and germinating conidia as well as germ tubes and hyphae of Aspergillus chevalieri, a xerophilic fungus predominant on moulded feed. Dormant conidia were not affected by 32 mM of ammonium propionate after a 28 h-treatment in demi water. Similar results were obtained with solely 0.52 mM MCFA. However, the combination of both components nearly eradicated formation of colonies from these conidia and was accompanied by distortion of the cellular structure as was visible with light- and transmission electron microscopy. Germination of conidia, characterised by swelling and germ tube formation, was significantly decreased in the presence of 16 mM ammonium propionate and 0.26 mM MCFA, while the latter component itself did not significantly decrease germination. We conclude that a combination of ammonium propionate and MCFA had a synergistic antifungal effect on dormant and germinating conidia. When the combination of ammonium propionate and MCFA was tested on hyphae for 30 min, we observed that cell death was significantly increased in comparison to components alone. Treatment of the hyphae with 16 mM of ammonium propionate caused aberrant mitochondria, as evidenced by irregularly shaped and enlarged mitochondria that contained electron-dense inclusions as observed by transmission electron microscopy. When the combination of ammonium propionate and MCFA was applied against the hyphae, more severe cell damage was observed, with signs of autophagy. Summarised, our results demonstrate synergistic antifungal effects of ammonium propionate and medium chain fatty acids on fungal survival structures, during their germination and after a short (sudden) treatment of growing cells. This is of potential importance for several areas of feed and food storage and shelf-life., Competing Interests: Declaration of competing interest The research is funded by Trouw Nutrition and Selko in order to investigate if a synergistic effect between ammonium propionate and MCFA exists., (Copyright © 2024. Published by Elsevier B.V.)

Background: Along bacteria, yeasts are common in forages and forage fermentations as spoilage microbes or as additives, yet few studies exist with species-level data on these fungi's occurrence in feedstuff. Active dry yeast and other yeast-based products are also common feed additives in animal husbandry. Here, we aimed to characterize both fermented and non-fermented milking cow feedstuff samples from Hungary to assess their microbial diversity in the first such study from Central Europe., Results: We applied long-read bacterial metabarcoding to 10 fermented and 25 non-fermented types of samples to assess bacterial communities and their characteristics, surveyed culturable mold and yeast abundance, and identified culturable yeast species. Fermented forages showed the abundance of Aerococcaceae, Bacillaceae, Brucellaceae, Lactobacillaceae, Staphylococcaceae, and Thermoactinomycetaceae, non-fermented ones had Cyanothecaceae, Enterobacteriaceae, Erwiniaceae, Gomontiellaceae, Oxalobacteraceae, Rhodobiaceae, Rickettsiaceae, and Staphylococcaceae. Abundances of bacterial families showed mostly weak correlation with yeast CFU numbers, only Microcoleaceae (positive) and Enterococcaceae and Alcaligenaceae (negative correlation) showed moderate correlation. We identified 14 yeast species, most commonly Diutina rugosa, Pichia fermentans, P. kudriavzevii, and Wickerhahomyces anomalus. We recorded S. cerevisiae isolates only from animal feed mixes with added active dry yeast, while the species was completely absent from fermented forages. The S. cerevisiae isolates showed high genetic uniformity., Conclusion: Our results show that both fermented and non-fermented forages harbor diverse bacterial microbiota, with higher alpha diversity in the latter. The bacterial microbiome had an overall weak correlation with yeast abundance, but yeasts were present in the majority of the samples, including four new records for forages as a habitat for yeasts. Yeasts in forages mostly represented common species including opportunistic pathogens, along with a single strain of Saccharomyces used as a feed mix additive., (© 2024. The Author(s).)

Background: The recurrent contaminations of feed materials with mycotoxigenic fungi can endanger both farmed animals and humans. Biosynthesized nanomaterials are assumingly the ideal agents to overcome fungal invasion in feed/foodstuffs, especially when utilizing sustainable sources for synthesis. Herein, the phycosynthesis of selenium nanoparticles (SeNPs) was targeted using Cystoseira myrica algal extract (CE), and the conjugation of CE/SeNPs with chitosan nanoparticles (NCt) to produce potential antifungal nanocomposites for controlling Aspergillus flavus isolates in fish feed., Results: The phycosynthesis of SeNPs with CE was effectually carried out and validated using visible/UV analysis, X-ray diffraction and transmission microscopy; CE/SeNPs had diameters of 8.7 nm and spherical shapes. NCt/CE/SeNPs nanocomposite (173.3 nm mean diameter) was achieved and the component interactions were validated via infrared spectroscopic analysis. The antifungal assessment of screened nanomaterials against three Aspergillus flavus strains indicated that NCt/CE/SeNPs exceeded the fluconazole action using qualitative/quantitative assays. Severe alteration/distortions in A. flavus mycelial structure and morphology were microscopically observed within 48 h of NCt/CE/SeNPs treatment. The treatment of feed ingredients (crushed corn and feed powder) by blending with nanomaterials (NCt, CE/SeNPs and NCt/CE/SeNPs) led to significant reduction in A. flavus count/growth after storage for 7 days; NCt/CE/SeNPs could completely inhibit any fungal growth in feed material., Conclusion: The pioneering phycosynthesis of CE/SeNPs and their nanoconjugation with NCt generated bioactive antifungal agents to control A. flavus strains. The innovatively constructed NCt/CE/SeNPs nanocomposite is advised for application as an effectual, biosafe and natural fungicidal conjugate for the protection of fish feed from mycotoxigenic fungi. © 2024 Society of Chemical Industry., (© 2024 Society of Chemical Industry.)

Zearalenone (ZEA) is a mycotoxin produced by Fusarium species, and cause contamination of food and feed, with impacts in animal production and in food production chain. Effective detoxifying methods, such as biodegradation, are therefore required. This study aimed to isolate microorganisms and screen ZEA detoxifying strains. As a result, 197 microorganisms were isolated, and six were initially selected after colorimetric screening. ZEA (1 µg/mL) was added to culture media, and after 24 h, all six microorganisms were able to degrade ZEA, without the formation of α-ZOL. One isolate eliminated ~ 99% of ZEA and was identified as Bacillus velezensis CL197. ZEA metabolites produced by the bacteria were evaluated, and no metabolites with greater or similar toxicity than ZEA were detected. This strain was applied to swine in vitro digestion, and up to 64% of ZEA was degraded. B. velezensis CL197 significantly degraded ZEA, demonstrating potential to be used as a detoxifying agent in the food production chain as a biocontrol agent., (© 2024. The Author(s).)

Background: Deoxynivalenol (DON) is a type B trichothecene mycotoxin that is commonly found in cereals and grains worldwide. The presence of this fungal secondary-metabolite raises public-health concerns at both the agriculture and food industry level. Recently, we have shown that DON has a negative impact on gut integrity, a feature also noticed for Campylobacter (C.) jejuni. We further demonstrated that DON increased the load of C. jejuni in the gut and inner organs. In contrast, feeding the less toxic DON metabolite deepoxy-deoxynivalenol (DOM-1) to broilers reduced the Campylobacter load in vivo. Consequently, it can be hypothesized that DON and DOM-1 have a direct effect on the growth profile of C. jejuni. The aim of the present study was to further resolve the nature of this interaction in vitro by co-incubation and RNA-sequencing., Results: The co-incubation of C. jejuni with DON resulted in significantly higher bacterial growth rates from 30 h of incubation onwards. On the contrary, the co-incubation of C. jejuni with DOM-1 reduced the CFU counts, indicating that this DON metabolite might contribute to reduce the burden of C. jejuni in birds, altogether confirming in vivo data. Furthermore, the transcriptomic profile of C. jejuni following incubation with either DON or DOM-1 differed. Co-incubation of C. jejuni with DON significantly increased the expression of multiple genes which are critical for Campylobacter growth, particularly members of the Flagella gene family, frr (ribosome-recycling factor), PBP2 futA-like (Fe 3+ periplasmic binding family) and PotA (ATP-binding subunit). Flagella are responsible for motility, biofilm formation and host colonization, which may explain the high Campylobacter load in the gut of DON-fed broiler chickens. On the contrary, DOM-1 downregulated the Flagella gene family and upregulated ribosomal proteins., Conclusion: The results highlight the adaptive mechanisms involved in the transcriptional response of C. jejuni to DON and its metabolite DOM-1, based on the following effects: (a) ribosomal proteins; (b) flagellar proteins; (c) engagement of different metabolic pathways. The results provide insight into the response of an important intestinal microbial pathogen against DON and lead to a better understanding of the luminal or environmental acclimation mechanisms in chickens., (© 2024. The Author(s).)

Probiotics are live microorganisms that, when administered in adequate quantities, provide health benefits to the host. In this study, phenotypic and genotypic methods were used to evaluate the probiotic properties of Bacillus altitudinis 1.4. The isolate was sensitive to all antimicrobials tested and presented a positive result in the hemolysis test. B. altitudinis 1.4 spores were more resistant than vegetative cells, when evaluated in simulation of cell viability in the gastrointestinal tract, as well as adhesion to the intestinal mucosa. The isolate was capable of self-aggregation and coaggregation with pathogens such as Escherichia coli ATCC 25922 and Salmonella Enteritidis ATCC 13076. Genomic analysis revealed the presence of genes with probiotic characteristics. From this study it was possible to evaluate the gene expression of pro-inflammatory and anti-inflammatory cytokines for different treatments. Viable vegetative cells of B. altitudinis 1.4 increased the transcription of pro-inflammatory factors, in addition to also increasing the transcription of IL-10, indicating a tendency to stimulate a pro-inflammatory profile. Given the results presented, B. altitudinis 1.4 showed potential to be applied in the incorporation of this microorganism into animal feed, since the spores could tolerate the feed handling and pelletization processes., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Lactic acid bacteria (LAB), particularly Lactobacilli strains, represent a widely studied and promising group of probiotics with numerous potential health benefits. In this study, we isolated LAB strains from fecal samples of healthy broiler chickens and characterized their probiotic properties. Out of 62 initial isolates, five strains were selected for further investigations based on their antibacterial activity against pathogenic bacteria. These selected strains were identified as Lactiplantibacillus species. They exhibited desirable probiotic traits, including non-hemolyis, non-cytotoxicity, lack of antibiotic resistance, acid tolerance, auto-aggregation, and antioxidative potential. Encapsulation of these strains in alginate beads enhanced their survival compared to free cells, in stomach (69-87 % vs. 34-47 %) and intestinal (72-100 % vs. 27-51 %) juices, after 120 min exposure. These findings suggest that encapsulated Lactiplantibacillus strains could be used as feed additives for broiler chickens. Nevertheless, further studies are needed to set on their probiotic potential in vivo., Competing Interests: Declaration of Competing Interest The authors declare no conflict of interest, (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Fungal and mycotoxin control at a primary stage in the food chain is crucial to maintaining the nutritional quality of animal feed. The control of fungal and mycotoxins is one of the essential points that a good biosecurity program must establish to ensure the safe feeding and protection of animal and human health. Acquiring a comprehensive understanding of the role of mycotoxins is vital to identifying breaches of this control and enabling the performance of proper risk assessments and accurate risk management strategies. This study focused on the identification of regulated and emerging mycotoxins in agricultural fields and dairy farms through an analytical methodology by ultra high-performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-QTrap-MS/MS). This allowed us to identify a Portuguese mycotoxin profile in the maize value chain for the first time. Regarding our occurrence data, mycotoxins were identified in all samples, namely seeds, flowering plants, grain and forage at harvest, maize silage, and animal feed. FBs and ENNB were highly present in all stages of the production cycle. This work offers an initial insight into a full screening of regulated and emerging mycotoxins within an important agricultural commodity. The high occurrence of these compounds points to the need to perform occurrence surveys from an integrative perspective to protect consumers' health, especially within food chains that provide various important staple foods worldwide.

One third of the food produced for human consumption is currently lost or wasted. Insects have a high potential for converting organic waste- and by-products into food and feed for a growing human population due to symbiosis with microorganisms. These symbioses provide an untapped reservoir of functional microbiomes that can be used to improve industrial insect production but are poorly studied in most insect species. Here we review the most current understanding and challenges of valorizing organic waste- and by-products through insects and their microbiomes for food and feed, and emerging novel food technologies that can be used to investigate and manipulate host(insects)-microbiome interactions. We further construct a holistic framework, by integration of novel food technologies including holo-omics, genome editing, breeding, phage therapy, and administration of prebiotics and probiotics to investigate and manipulate host(insects)-microbiome interactions, and solutions for achieving stakeholder acceptance of novel food technologies for a sustainable food production., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Pet food have been considered as possible vehicles of bacterial pathogens. The sudden boom of the pet food industry due to the worldwide increase in companion animal ownership calls for pet food investigations. Herein, this study aimed to determine the frequency, antimicrobial susceptibility profile, and molecular characteristics of coagulase-negative staphylococci (CoNS) in different pet food brands in Brazil. Eighty-six pet food packages were screened for CoNS. All isolates were identified at species level by MALDI-TOF MS and species-specific PCR. Antimicrobial susceptibility testing was performed by disc diffusion and broth microdilution (vancomycin and teicoplanin only) methods. The D-test was used to screen for inducible clindamycin phenotype (MLS-B). SCCmec typing and detection of mecA, vanA, vanB, and virulence-encoding genes were done by PCR. A total of 16 (18.6 %) CoNS isolates were recovered from pet food samples. Isolates were generally multidrug-resistant (MDR). All isolates were completely resistant (100 %) to penicillin. Resistances (12.5 % - 75 %) were also observed for fluoroquinolones, sulfamethoxazole-trimethoprim, tetracycline, rifampicin, erythromycin, and tobramycin. Isolates were susceptible to vancomycin (MICs <0.25-1 μg/mL) and teicoplanin (MICs <0.25-4 μg/mL). Intriguingly, 3/8 (37.5 %) CoNS isolates with the ERY R CLI S antibiotype expressed MLS-B phenotype. All isolates harboured blaZ gene. Seven (43.8 %) isolates carried mecA; and among them, the SCCmec Type III was the most frequent (n = 5/7; 71.4 %). Isolates also harboured seb, see, seg, sej, sem, etb, tsst, pvl, and hla toxin virulence-encoding genes (6.3 % - 25 %). A total of 12/16 (75 %) isolates were biofilm producers, while the icaAB gene was detected in an S. pasteuri isolate. Herein, it is shown that pet food is a potential source of clinically important Gram-positive bacterial pathogens. To the best of our knowledge, this is the first report of MLS-B phenotype and MR-CoNS in pet food in Latin America., Competing Interests: Declaration of competing interest A.C.G. has recently received research funding and/or consultation fees from bioMerieux Eurofarma, MSD, Pfizer, Roche, Sandoz, União Química, and United Medical. Other authors have nothing to declare. This study was not financially supported by any Diagnostic/Pharmaceutical company., (Copyright © 2024 Elsevier B.V. All rights reserved.)

BackgroundThe pet industry is expanding worldwide, particularly raw meat-based diets (RMBDs). There are concerns regarding the safety of RMBDs, especially their potential to spread clinically relevant antibiotic-resistant bacteria or zoonotic pathogens.AimWe aimed to investigate whether dog food, including RMBD, commercially available in Portugal can be a source of Salmonella and/or other Enterobacteriaceae strains resistant to last-line antibiotics such as colistin.MethodsFifty-five samples from 25 brands (21 international ones) of various dog food types from 12 suppliers were screened by standard cultural methods between September 2019 and January 2020. Isolates were characterised by phenotypic and genotypic methods, including whole genome sequencing and comparative genomics.ResultsOnly RMBD batches were contaminated, with 10 of 14 containing polyclonal multidrug-resistant (MDR) Escherichia coli and one MDR Salmonella . One turkey-based sample contained MDR Salmonella serotype 1,4,[5],12:i:- ST34/cgST142761 with similarity to human clinical isolates occurring worldwide. This Salmonella exhibited typical antibiotic resistance ( bla TEM  +  strA-strB  +  sul2  +  tet(B) ) and metal tolerance profiles ( pco  +  sil  +  ars ) associated with the European epidemic clone. Two samples (turkey/veal) carried globally dispersed MDR E. coli (ST3997-complexST10/cgST95899 and ST297/cgST138377) with colistin resistance (minimum inhibitory concentration: 4 mg/L) and mcr-1 gene on IncX4 plasmids, which were identical to other IncX4 circulating worldwide.ConclusionSome RMBDs from European brands available in Portugal can be a vehicle for clinically relevant MDR Salmonella and pathogenic E. coli clones carrying genes encoding resistance to the last-line antibiotic colistin. Proactive actions within the One Health context, spanning regulatory, pet-food industry and consumer levels, are needed to mitigate these public health risks.

Water kefir is a sparkling, slightly acidic fermented beverage made from sugar, water, and water kefir grains, which are a mixture of yeast and bacteria. These grains produce a variety of fermentation compounds such as lactic acid, acetaldehyde, acetoin, ethanol and carbon dioxide. In this study, a high-throughput sequencing technique was used to characterize the bacterial composition of the original water kefir from which potential probiotics were obtained. We studied the bacterial diversity of both water kefir grains and beverages. DNA was extracted from three replicate samples of both grains and beverages using the Powerlyzer Microbial Kit. The hypervariable V1-V2 region of the bacterial 16S ribosomal RNA gene was amplified to prepare six DNA libraries. Between 1.4M and 2.4M base-pairs were sequenced for the library. In total, 28721971 raw reads were obtained from all the samples. Estimated species richness was higher in kefir beverage samples compared to grain samples. Moreover, a higher level of microbial alpha diversity was observed in the beverage samples. Particularly, the predominant bacteria in beverages were Anaerocolumna and Ralstonia, while in grains Liquorilactobacillus dominated, with lower levels of Leuconostoc and Oenococcus. Although the bacterial diversity in kefir grains was low because only three genera were the most represented, all of them are LAB bacteria with the potential to serve as probiotics in the artificial feeding of bees., (Copyright © 2024 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.)

Mycotoxins present a significant health concern within the animal-feed industry, with profound implications for the pig-farming sector. The objective of this study was to evaluate the efficacy of two commercial adsorbents, an organically modified clinoptilolite (OMC) and a multicomponent mycotoxin detoxifying agent (MMDA), to ameliorate the combined adverse effects of dietary aflatoxins (AFs: sum of AFB 1 , AFB 2 , AFG 1 , and AFG 2 ), fumonisins (FBs), and zearalenone (ZEN) at levels of nearly 0.5, 1.0, and 1.0 mg/kg, on a cohort of cross-bred female pigs ( N = 24). Pigs were randomly allocated into six experimental groups (control, mycotoxins (MTX) alone, MTX + OMC 1.5 kg/ton, MTX + OMC 3.0 kg/ton, MTX + MMDA 1.5 kg/ton, and MTX + MMDA 3.0 kg/ton), each consisting of four individuals, and subjected to a dietary regimen spanning 42 days. The administration of combined AFs, FBs, and ZEN reduced the body-weight gain and increased the relative weight of the liver, while there was no negative influence observed on the serum biochemistry of animals. The supplementation of OMC and MMDA ameliorated the toxic effects, as observed in organ histology, and provided a notable reduction in residual AFs, FBs, and ZEN levels in the liver and kidneys. Moreover, the OMC supplementation was able to reduce the initiation of liver carcinogenesis without any hepatotoxic side effects. These findings demonstrate that the use of OMC and MMDA effectively mitigated the adverse effects of dietary AFs, FBs, and ZEN in piglets. Further studies should explore the long-term protective effects of the studied adsorbent supplementation to optimize mycotoxin management strategies in pig-farming operations.

In recent years, there has been an increased interest in beef cattle shedding of foodborne pathogens due to the potential to contaminate surrounding food crops; however, the number of studies published on this topic has declined as the majority of research has emphasized on postharvest mitigation efforts. A field study was conducted to determine the prevalence of pathogens and indicator bacteria in beef cattle fed two different direct-fed microbials (DFMs). Fecal samples from a total of 3,708 crossbred yearling cattle randomly assigned to 16 pens and two treatment groups at a commercial cattle feedlot were taken. During the study period, diets were supplemented with two different DFMs i.) Lactobacillus acidophilus (NP51) and Propionibacterium freudenreichii (NP24) (9 log 10 CFU/head/day), and ii.) Lactobacillus salivarius (L28) (6 log 10 CFU/head/day). Fecal samples from pen floors were collected on days 0, 21, 42, 63, 103, and analyzed for the presence of Salmonella and E. coli O157:H7 and concentration of E. coli O157:H7, Enterobacteriaceae, and C. perfringens. Fecal samples collected from cattle fed L28 had significantly lower concentration of C. perfringens (p < 0.05) and had a similar prevalence with no significant differences in E. coli O157:H7 as those fed NP51/NP24 through the study until day 103. On day 103, the prevalence in cattle fed L28 was 40% with a concentration of 0.95 log 10 MPN/g while those fed NP51/NP24 were 65% with a concentration of 1.2 log 10 MPN/g. Cattle supplemented with NP51/NP24 achieved a significant log reduction of EB by 2.4 log 10 CFU/g over the course of the 103-day supplementation period compared to L28. Salmonella prevalence was also measured, but not detected in any samples at significant amounts to draw conclusions. It is evident that E. coli O157:H7 and other foodborne pathogens are still prevalent in cattle operations and that preharvest mitigation strategies should be considered to reduce the risk to beef products., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Drs. Brashears and Nightingale are co-founders and own shares of NexGen Innovations, LLC and their participation is governed by a management plan in place at Texas Tech University to mitigate risks from conflicts of interest. Dr. Brashears receives royalty income from the sales of Bovamine Defend., (Copyright © 2023. Published by Elsevier Inc.)

Mycotoxin-contaminated feeds may negatively affect broiler chickens’ health; hence, a sustainable approach to achieve mycotoxin elimination is necessary. This study aimed to evaluate the efficacy of red yeast (Sporidiobolus pararoseus; RY) as a novel mycotoxin binder in broilers. A total of 1440 one-week-old male broiler chicks were randomly assigned to 12 treatments in a 3 × 4 factorial design. The dietary treatments included three levels of mycotoxin-contaminated diets (0 µg kg−1 (0% of mycotoxin; MT), 50 µg kg−1 (50% MT), and 100 µg kg−1 (100% MT)) and four levels of mycotoxin binders (0.0 and 0.5 g kg−1 commercial binder, and 0.5 and 1.0 g kg−1 RY). Experimental diets were contaminated with aflatoxin B1, zearalenone, ochratoxin A, T-2 toxin, and deoxynivalenol in the basal diet. Furthermore, the parameters including feed intake, body weight, and mortality rate were recorded on a weekly basis. After feeding for 28 days, blood and organ samples were collected randomly to determine the blood biochemistry, relative organ weights, and gut health. The results indicated that mycotoxin-contaminated diets reduced the average daily weight gain (ADG), villus height (VH), and villus height per the crypt depth ratio (VH:CD) of the intestine, as well as the population of Lactobacillus sp. and Bifidobacterium sp. in the cecal (p < 0.05), whereas they increased the mycotoxins concentration in the blood samples and the apoptosis cells (TUNEL positive) in the liver tissue (p < 0.01) of broiler chicken. In contrast, RY-supplemented diets had better ADG values and lower chicken mortality rates (p < 0.05). Moreover, these combinations positively impacted the relative organ weights, blood parameters, bacteria population, intestinal morphology, and pathological changes in the hepatocytes (p < 0.05). In conclusion, RY supplementation effectively alleviated the toxicity that is induced by AFB1 and OTA, mainly, and could potentially be applied as a novel feed additive in the broiler industry.

Mycotoxins are toxic secondary metabolites produced by a variety of fungi, which when ingested can cause several deleterious effects to the health of humans and animals. In this work, the detection and quantification of six major mycotoxins (aflatoxins-AFLA, deoxynivalenol-DON, fumonisins-FUMO, ochratoxin A-OTA, T-2 toxin-T-2 and zearalenone-ZON) in 1749 samples of feed and feed ingredients for cattle, collected in Brazil between 2017 and 2021, was carried out using enzyme-linked immunosorbent assay (ELISA). In total, 97% of samples were contaminated with at least one mycotoxin, yet, very few samples exceeded the lowest European Union guidance values for cattle, and the estimated daily intake also showed a low risk for the animals. However, co-occurrences were widely observed, as 87% of samples contained two or more mycotoxins at the same time, and the presence of more than one mycotoxin at the same time in feed can lead to interactions. In conclusion, the contamination of feed and feed ingredients for cattle with mycotoxins in Brazil is very common. Hence, the monitoring of these mycotoxins is of significant importance for food safety.

Aims: Bacillus cereus is often responsible for foodborne diseases and both local and systemic infections in humans. Cases of infection in other mammals are rather rare. In this study, we report a B. cereus feed-related outbreak that caused the death of 6234 pigs in Italy., Methods and Results: Massive doses of a Gram-positive, spore-forming bacterium were recovered from the animal feed, faeces of survived pigs and intestinal content of dead ones. The B. cereus MM1 strain was identified by MALDI-TOF MS and typified by RAPD-PCR. The isolate was tested for the production of PC-PLC, proteases, hemolysins and biofilm, for motility, as well as for the presence of genes encoding tissue-degrading enzymes and toxins. Antimicrobial resistance and pathogenicity in Galleria mellonella larvae were also investigated. Our results show that the isolated B. cereus strain is swimming-proficient, produces PC-PLC, proteases, hemolysins, biofilm and carries many virulence genes. The strain shows high pathogenicity in G. mellonella larvae., Conclusions: The isolated B. cereus strain demonstrates an aggressive profile of pathogenicity and virulence, being able to produce a wide range of determinants potentially hazardous to pigs' health., Significance and Impact of Study: This study highlights the proficiency of B. cereus to behave as a devastating pathogen in swine if ingested at high doses and underlines that more stringent quality controls are needed for livestock feeds and supplements., (© 2022 The Authors. Journal of Applied Microbiology published by John Wiley & Sons Ltd on behalf of Society for Applied Microbiology.)

Objective: To assess the potential contamination of commercial raw dog food products with bacteria of the Enterobacterales order that produce extended spectrum beta-lactamase (ESBL) and carbapenemase enzymes, determine risk factors for contamination, and understand isolate genetic diversity., Samples: A total of 200 canine raw food products., Methods: Products were cultured on selective chromogenic agar following enrichment steps. Whole-genome sequencing was performed for isolates that were confirmed to produce an ESBL. Isolates were characterized by antimicrobial resistance genes, and multilocus sequences typing, and compared to other isolates in the NCBI database for clonality. Preservation method and protein sources were assessed as potential risk factors for contamination with ESBL and carbapenemase-producing bacteria of the Enterobacterales order., Results: No carbapenemase-producing Enterobacterales (CPE) were identified, but ESBL-producing Enterobacterales bacteria were isolated from 20/200 products (10.0%; 95% CI, 7.3 to 16.5%), all of which were frozen. Pork-derived protein source products were 8.1 times (P = .001; 95% CI, 2.53 to 26.2) more likely to carry ESBL-producing Enterobacterales bacteria than other protein sources. WGS analysis confirmed the presence of ESBL genes in a total of 25 distinct isolates (19 Escherichia coli, 5 Klebsiella pneumoniae, and 1 Citrobacter braakii). Genes encoding CTX-M type ESBL enzymes were the most common (24/25 isolates, 96.0%) with blaCTX-M-27 being the most common allele (8/25, 32.0%)., Clinical Relevance: Frozen, raw food products may serve as a route of transmission of ESBL-producing Enterobacterales bacteria to companion animals. Veterinarians should advise owners about the risks of raw food diets, including potential exposure to antimicrobial-resistant bacteria.

This study aimed to evaluate the effects of Saccharomyces cerevisiae and Megasphaera elsdenii as direct fed microbials (DFM) in beef cattle finishing diets to alleviate acute ruminal lactic acidosis in vitro. A dual-flow continuous culture system was used. Treatments were a Control, no DFM; YM1, S. cerevisiae and M. elsdenii strain 1; YM2, S. cerevisiae and M. elsdenii strain 2; and YMM, S. cerevisiae and half of the doses of M. elsdenii strain 1 and strain 2. Each DFM dose had a concentration of 1 × 10 8  CFU/mL. Four experimental periods lasted 11 days each. For the non-acidotic days (day 1-8), diet contained 50:50 forage to concentrate ratio. For the challenge days (day 9-11), diet contained 10:90 forage to concentrate ratio. Acute ruminal acidosis was successfully established. No differences in pH, D-, L-, or total lactate were observed among treatments. Propionic acid increased in treatments containing DFM. For N metabolism, the YMM treatment decreased protein degradation and microbial protein synthesis. No treatment effects were observed on NH 3 -N concentration; however, efficiency of N utilization by ruminal bacteria was greater than 80% during the challenge period and NH 3 -N concentration was reduced to approximately 2 mg/dL as the challenge progressed., (© 2022. The Author(s).)

Food for human and animal consumption can provide a vehicle for the transfer of pathogenic and antimicrobial-resistant bacteria into the food chain. We investigated the antimicrobial susceptibility of 453 Salmonella isolates collected from raw feed components, equipment and finished feed from 17 commercial feed mills in Australia between 2012 and 2021. Previous studies have found Salmonella prevalence and the diversity of Salmonella serotypes are greatest in the raw feed components. We, therefore, hypothesised that we would find a greater proportion of antimicrobial-resistant Salmonella isolates in the raw feed components compared to other sample types. We found that of 453 isolates tested, 356 (0.80) were susceptible to all antimicrobials tested, 49 (0.11) were nonsusceptible to streptomycin only and 48 (0.11) were resistant to two or more antimicrobials. Of the 48 antimicrobial-resistant isolates, 44 were found in feed milling equipment, two in raw feed components and two in finished feed. Statistical analysis, using a logistic regression with random effects model, found that the population-adjusted mean probability of detecting antimicrobial-resistant Salmonella isolates from feed milling equipment of 0.39, was larger than the probability of detecting resistant isolates in raw feed components 0.01, (P < 0.001) and in finished feed, 0.11, (P = 0.006). This propensity for antimicrobial-resistant bacteria to colonise feed milling equipment has not been previously reported. Further studies are required to understand the ecology of antimicrobial-resistant Salmonella in the feed milling environment., (© 2022 The Authors. Australian Veterinary Journal published by John Wiley & Sons Australia, Ltd on behalf of Australian Veterinary Association.)

Mycotoxins are a group of toxic secondary metabolites produced in the food chain by fungi through the infection of crops both before and after harvest. Mycotoxins are one of the most important food safety concerns due to their severe poisonous and carcinogenic effects on humans and animals upon ingestion. In the last decade, insects have received wide attention as a highly nutritious, efficient and sustainable source of animal-derived protein and caloric energy for feed and food purposes. Many insects have been used to convert food waste into animal feed. As food waste might contain mycotoxins, research has been conducted on the metabolism and detoxification of mycotoxins by edible insects. The mycotoxins that have been studied include aflatoxins, fumonisins, zearalenone (ZEN), vomitoxin or deoxynivalenol (DON), and ochratoxins (OTAs). Aflatoxin metabolism is proved through the production of hydroxylated metabolites by NADPH-dependent reductases and hydroxylases by different insects. ZEN can be metabolized into α- and β-zearalenol. Three DON metabolites, 3-, 15-acetyl-DON, and DON-3-glucoside, have been identified in the insect DON metabolites. Unfortunately, the resulting metabolites, involved enzymes, and detoxification mechanisms of OTAs and fumonisins within insects have yet to be identified. Previous studies have been focused on the insect tolerance to mycotoxins and the produced metabolites; further research needs to be conducted to understand the exact enzymes and pathways that are involved.

Asterarcys quadricellulare (AQ) is a microalgal species with potential applications in improving the quality of animal feed, and safety studies on this species are lacking. Therefore, this study presents safety data on an industrially cultivated strain of AQ tested using the following Organisation for Economic Co-operation and Development (OECD) guidelines: acute skin irritation in rabbits; skin sensitisation in guinea pigs; acute eye irritation in rabbits; acute oral fixed-dose procedure in rats; and bacterial reverse mutation using the B.N. Ames technique. Results showed that AQ is non-irritant and non-sensitising to skin. AQ caused transient conjunctival lacrimation and redness; however, the scores for these clinical signs translated into low ocular irritation indices and classification of AQ as non-irritant to the eyes. An acute oral dose of AQ (2000 mg/kg) did not cause mortality, change in body weight gain, or any general, functional, and neurobehavioral clinical signs. In five strains of Salmonella typhimurium bacteria, treatment with AQ did not cause biologically or statistically significant changes in the number of revertant colonies, indicating that AQ does not cause mutagenic toxicity. This study demonstrates the safety of a heterotrophically-produced strain of AQ and supports its use as a safe and non-toxic feed ingredient., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Feeding experiments with juvenile grass carp ( Ctenopharyngodon idella ) fed with genetically modified maize MON 810 or DAS-59122 dried leaf biomass were carried out with 1-, 3- and 6-month exposures. Dosages of 3-7 μg/fish/day Cry1Ab or 18-55 μg/fish/day Cry34Ab1 toxin did not cause mortality. No difference occurred in body or abdominal sac weights. No differences appeared in levels of inorganic phosphate, calcium, fructosamine, bile acids, triglycerides, cholesterol, and alanine and aspartame aminotransferases. DAS-59122 did not alter blood parameters tested after 3 months of feeding. MON 810 slightly decreased serum albumin levels compared to the control, only in one group. Tapeworm ( Bothriocephalus acheilognathi ) infection changed the levels of inorganic phosphate and calcium. Cry34Ab1 toxin appeared in blood (12.6 ± 1.9 ng/mL), but not in the muscle. It was detected in B. acheilognathi. Cry1Ab was hardly detectable in certain samples near the limit of detection. Degradation of Cry toxins was extremely quick in the fish gastrointestinal tract. After 6 months of feeding, only mild indications in certain serum parameters were observed: MON 810 slightly increased the level of apoptotic cells in the blood and reduced the number of thrombocytes in one group; DAS-59122 mildly increased the number of granulocytes compared to the near-isogenic line.

This study addresses an advantageous application of a urinary zearalenone (ZEN) monitoring system not only for surveillance of ZEN exposure at the production site of breeding cows but also for follow-up monitoring after improvement of feeds provided to the herd. As biomarkers of effect, serum levels of the anti-Müllerian hormone (AMH) and serum amyloid A (SAA) concentrations were used. Based on the results of urinary ZEN measurement, two cows from one herd had urinary ZEN concentrations which were two orders of magnitude higher (ZEN: 1.34 mg/kg, sterigmatocystin (STC): 0.08 mg/kg in roughages) than the levels of all cows from three other herds (ZEN: not detected, STC: not detected in roughages). For the follow-up monitoring of the herd with positive ZEN and STC exposure, urine, blood, and roughage samples were collected from five cows monthly for one year. A monitoring series in the breeding cattle herd indicated that feed concentrations were not necessarily reflected in urinary concentrations; urinary monitoring assay by ELISA may be a simple and accurate method that reflects the exposure/absorption of ZEN. Additionally, although the ZEN exposure level appeared not to be critical compared with the Japanese ZEN limitation in dietary feeds, a negative regression trend between the ZEN and AMH concentrations was observed, indicating that only at extremely universal mycotoxin exposure levels, ZEN exposure may affect the number of antral follicles in cattle. A negative regression trend between the ZEN and SAA concentrations could also be demonstrated, possibly indicating the innate immune suppression caused by low-level chronic ZEN exposure. Finally, significant differences ( p = 0.0487) in calving intervals between pre-ZEN monitoring (mean ± SEM: 439.0 ± 41.2) and post-ZEN monitoring (349.9 ± 6.9) periods were observed in the monitored five cows. These preliminary results indicate that the urinary ZEN monitoring system may be a useful practical tool not only for detecting contaminated herds under field conditions but also provides an initial look at the effects of long-term chronic ZEN/STC (or other co-existing mycotoxins) exposure on herd productivity and fertility.

Background: Feed shortage is a factor restricting animal production in the tropics, therefore how to use natural woody plant resources as animal feed is an important strategy., Results: Under the dual stress of an anaerobic and acidic environment, the microbial response during the fermentation of paper mulberry (PM) silage was found to be sensitive. The Gram-negative bacteria and mould died, and the dominant microbial community rapidly shifted to Gram-positive bacteria, resulting in a large reduction in microbial diversity and abundance. Exogenous bran additives interfered with the stress effects of the woody silage environment. Wheat bran (WB) accelerated the response of microorganisms to the anaerobic stress, and lactic acid bacteria became the dominant microbial community, thereby enhancing the lactic acid fermentation of silage, affecting the metabolic pathways of microorganisms, and improving the flavour and quality of the silage. Addition of rice bran made Enterobacter and Clostridium species quickly respond to the stress of the silage environment and become the predominant bacterial groups. In particular, anaerobic and spore-forming Clostridium species showed a strong tolerance to the silage environment, leading to butyric acid fermentation and protein degradation of the silage, and reducing its fermentation quality., Conclusion: The PacBio single-molecule real-time (SMRT) sequencing technology accurately revealed the microbial co-occurrence network and fermentation mechanism of silage. Our results indicate that PM can be used in combination with WB to prepare high-quality silage for animal production. © 2021 Society of Chemical Industry., (© 2021 Society of Chemical Industry.)

&lt;b&gt;Background and Objectives:&lt;/b&gt; Blue swimming crab, &lt;i&gt;Portunus pelagicus&lt;/i&gt; (Linnaeus, 1758) are economically important sources of soft-shell products for export demand. However, increasing over-exploitation and pollution towards the sea resources affected the production of the &lt;i&gt;P. pelagicus&lt;/i&gt;. The study aimed to determine the microalgal preference and feeding density of the monospecific (&lt;i&gt;Nannochloropsis oculata&lt;/i&gt;, &lt;i&gt;Chlorella vulgaris&lt;/i&gt;, &lt;i&gt;Isochrysis galbana&lt;/i&gt; and &lt;i&gt;Chaetoceros wighamii &lt;/i&gt;) by individual &lt;i&gt;P. pelagicus &lt;/i&gt;larvae. &lt;b&gt;Materials and Methods:&lt;/b&gt; In the microalgal preference study, one polystyrene tray with 100 holes was set up in the experimental tank (200 L) to place the 50 mL centrifuges tube. For feeding density study, the survival rate, Specific Growth Rate (SGR), Body Weight (BW) and Larvae Stage Index (LSI) of each larvae stage were collected from Zoea 1 (Z&lt;sub&gt;1&lt;/sub&gt;) till megalopa. &lt;b&gt;Results:&lt;/b&gt; Results for mean microalgal preference of individual larvae preferred most &lt;i&gt;Chlorella vulgaris&lt;/i&gt; during Zoea 1 and 2, the larvae consumed the highest of &lt;i&gt;Nannochloropsis&lt;/i&gt; &lt;i&gt;oculata&lt;/i&gt;. For Zoea 3 and 4, the highest preference is &lt;i&gt;Isochrysis galbana&lt;/i&gt;. For feeding regime, Zoea 1, larvae fed rotifer, &lt;i&gt;Artemia&lt;/i&gt; and &lt;i&gt;Chlorella vulgaris&lt;/i&gt; (T&lt;sub&gt;3&lt;/sub&gt;), was the highest survival rate, Zoea 2 was larvae fed rotifer, &lt;i&gt;Artemia&lt;/i&gt; and &lt;i&gt;Nannochloropsis oculata&lt;/i&gt; (T&lt;sub&gt;5&lt;/sub&gt;) and both Zoea 3 and Zoea 4, larvae fed rotifer, &lt;i&gt;Artemia&lt;/i&gt; and &lt;i&gt;Isochrysis galbana&lt;/i&gt; (T&lt;sub&gt;4&lt;/sub&gt;) was the highest survival rate. The highest SGR and LSI were obtained in T&lt;sub&gt;4&lt;/sub&gt; cultured in 14 days. &lt;b&gt;Conclusion:&lt;/b&gt; In conclusion, the microalgal preference and feeding density of the selected microalgae are different depending on each larvae stage.

Black soldier fly larvae (BSFL; Hermetia illucens) are promising insects for the conversion of organic waste streams into valuable biomolecules. Such waste streams can contain foodborne pathogens. To assess this risk factor, this study evaluated the presence of Staphylococcus aureus in waste streams as a substrate ingredient for BSFL production as well as in the rearing process. First, the general microbiological quality and the occurrence of S. aureus were investigated for different waste streams. Staphylococcus aureus was abundantly present. Control of pH and water activity should avoid pathogens, which cannot grow in single-substrate ingredients, redeveloping when mixing streams for optimal substrate conditions for BSFL production. Next, it was investigated whether S. aureus present in the substrate was ingested and/or eradicated by BSFL. In inoculation trials, with S. aureus added to chicken feed as the substrate at 3 or 7 log CFU/g, the larvae showed a reducing effect on S. aureus. After 6 days, S. aureus counts were below the detection limit (2.0 log CFU/g) in all larvae samples and decreased in the substrate to <2.0 and <3.1 log CFU/g for inoculation levels of 3 and 7 log CFU/g, respectively. While this is promising, it is still recommended to monitor and control this pathogen in BSFL rearing. Intriguingly, screening of antimicrobial activity of dominant microorganisms associated with BSFL showed a clear activity of Trichosporon isolates against S. aureus. Future research should explore whether Trichosporon , which is frequently observed in BSFL, plays a role in controlling specific microorganisms, such as S. aureus. IMPORTANCE Given the increasing need for (more sustainable) methods to upcycle organic waste streams, the interest to rear insects, like black soldier fly larvae (BSFL), on such streams is increasing. This study reveals that S. aureus is abundantly present in such waste streams, which might be a point of attention for insect producers. At the same time, it reveals that when S. aureus was inoculated in chicken feed as the substrate, it was not detected in the larvae and was reduced in the substrate after 6 days. Future inoculation trials should investigate whether this reduction is substrate dependent or not. Toward the future, the role of the BSFL microbiota in controlling intestinal bacterial community homeostasis should be explored, because one of the dominant microorganisms associated with BSFL, Trichosporon spp., showed clear activity against S. aureus.

The outbreak of diseases leading to substantial loss is a major bottleneck in aquaculture. Over the last decades, the concept of using feed probiotics was more in focus to address the growth and health of cultivable aquatic organisms. The objective of this review is to provide an overview of the distinct functionality of archaea from conventional probiotics in nutrient utilization, specific caloric contribution, evading immune response and processing thermal resistance. The prime limitation of conventional probiotics is the viability of desired microbes under harsh feed processing conditions. To overcome the constraints of commercial probiotics pertaining to incompatibility towards industrial processing procedure, a super microbe, archaea, appears to be a potential alternative approach in aquaculture. The peculiarity of the archaeal cell wall provides them with heat stability and rigidity under industrial processing conditions. Besides, archaea being one of the gut microbial communities participates in various health-oriented biological functions in animals. Thus, the current review devoted that administration of archaea in aquafeed could be a promising strategy in aquaculture. Archaea may be used as a potential probiotic with the possible modes of functions and advantages over conventional probiotics in aquafeed preparation. The present review also provides the challenges associated with the use of archaea for aquaculture and a brief outline of the patents on archaea to highlight the various use of archaea in different sectors., (© 2021. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Rhizopus oryzae is a fungus used to ferment tempeh in Indonesia and is generally recognized as safe (GRAS) for human consumption by the USA FDA. We previously assessed the effect of a tempeh extract on cortisol levels in zebrafish but did not include behavioral studies. Here, we measured the GABA content in three strains of Rhizopus oryzae , two isolated by us (MHU 001 and MHU 002) and one purchased. We then investigated the effect of tempeh on cortisol and the gut microbiota in a zebrafish experimental model. GABA concentration was the highest in MHU 002 (9.712 ± 0.404 g kg -1 ) followed by our MHU 001 strain and the purchased one. The fish were divided into one control group fed a normal diet and three experimental groups fed soybean tempeh fermented with one of the three strains of Rhizopus oryzae . After two weeks, individual fish were subjected to unpredicted chronic stress using the novel tank diving test and the tank light-dark test. Next-generation sequencing was used to analyze gut microbial communities and RT-PCR to analyze the expression of BDNF (brain-derived neurotrophic factor) gene and of other genes involved in serotonin signaling/metabolism in gut and brain. Tempeh-fed zebrafish exhibited increased exploratory behavior (less stress) in both tank tests. They also had significantly reduced gut Proteobacteria (include E. coli ) (51.90% vs. 84.97%) and significantly increased gut Actinobacteria (include Bifidobacterium spp.) (1.80% vs. 0.79%). The content of Bifidobacterium adolescentis , a "psychobiotic", increased ten-fold from 0.04% to 0.45%. Tempeh also increases BDNF levels in zebrafish brain. Rhizopus oryzae MHU 001 greatly improved the anti-stress effect of tempeh and microbiota composition in zebrafish gut.

Mycotoxins can be found in many foods consumed by humans and animals. These substances are secondary metabolites of some fungi species and are resistant to technological processes (cooking, frying, baking, distillation, fermentation). They most often contaminate products of animal (beef, pork, poultry, lamb, fish, game meat, milk) and plant origin (cereals, processed cereals, vegetables, nuts). It is estimated that about 25% of the world's harvest may be contaminated with mycotoxins. These substances damage crops and may cause mycotoxicosis. Many mycotoxins can be present in food, together with mold fungi, increasing the exposure of humans and animals to them. In this review we characterized the health risks caused by mycotoxins found in food, pet food and feed. The most important groups of mycotoxins are presented in terms of their toxicity and occurrence.

The contamination of food and feeds with mycotoxins poses a global health risk to humans and animals, with major economic consequences. Good agricultural and manufacturing practices can help control mycotoxin contamination. Since these actions are not always effective, several methods of decontamination have also been developed, including physical, chemical, and biological methods. Biological decontamination using microorganisms has revealed new opportunities. However, these biological methods require legal regulations and more research before they can be used in food production. Currently, only selected biological methods are acceptable for the decontamination of feed. This review discusses the literature on the use of microorganisms to remove mycotoxins and presents their possible mechanisms of action. Special attention is given to Saccharomyces cerevisiae yeast and lactic acid bacteria, and the use of yeast cell wall derivatives.

The present study describes genome annotation and phenotypic characterization of Bacillus velezensis ZBG17 and evaluation of its performance as antibiotic growth promoter substitute in broiler chickens. ZBG17 comprises 3.89 Mbp genome with GC content of 46.5%. ZBG17 could tolerate simulated gastrointestinal juices prevalent in the animal gut. Some adhesion-associated genomic features of ZBG17 supported the experimentally determined cell surface hydrophobicity and cell aggregation results. ZBG17 encoded multiple secondary metabolite gene clusters correlating with its broad-spectrum antibacterial activity. Interestingly, ZBG17 completely inhibited Salmonella enterica and Escherichia coli within 6 h and 8 h in liquid co-culture assay, respectively. ZBG17 genome analysis did not reveal any genetic determinant associated with reported safety hazards for use as a poultry direct-fed microbial. Dietary supplementation of ZBG17 significantly improved feed utilization efficiency and humoral immune response in broiler chickens, suggesting its prospective application as a direct-fed microbial in broiler chickens., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

The study aimed to investigate the efficacy of new mycotoxin adsorbents based on purified and activated bentonites combined with yeast and phytogenic compounds in fattening pigs. The experiment involved 96 pigs (31.2±2.4 kg). Control (C) group was fed a diet naturally contaminated with mycotoxins (5 mg/kg deoxynivalenol, DON) without an adsorbent. Treated groups received the feed with mycotoxin adsorbents: purified and activated bentonite (T1), purified and activated bentonite, yeast derivatives, phytogenic substances (T2), and purified, activated, and sulphurated bentonite with phytogenic substances (T3). Evaluated parameters involved growth performance, organ weight, small intestine and liver histopathology, complete blood count, serum biochemistry, antioxidant status of the organism and total and free DON content in urine. In all treated groups, an significant increase in intestinal GSH and GSH/GSSG ratio was observed when compared to C. No significant effects on liver and kidney weight, complete blood count, serum or intestinal malondialdehyde concentration, or total/free DON content in urine were observed. All adsorbents improved histopathological findings in the liver when compared to C. Moreover, T1, and T2 groups showed no presence of inflammatory reaction or necrotic changes in the livers. Although, mycotoxin adsorbents investigated in this study had no significant impact on pig growth performance, they reduced the oxidative stress, and on the tissue level they protected the jejunal tissue and liver parenchyma under deoxynivalenol challenge., Competing Interests: The authors have declared that no competing interests exist.

Clostridium perfringens (CP) is the principal pathogenic bacterium of chicken necrotic enteritis (NE), which causes substantial economic losses in poultry worldwide. Although probiotics are known to provide multiple benefits, little is known about the potential effects of Bacillus subtilis ( B . subtilis ) application in preventing CP-induced necrotic enteritis. In this study, 450 male Arbor Acres broilers were divided into 5 experimental treatments: A: basal diet (control group); B: basal diet and CP challenge (model group); C: CP challenge+10 mg/kg enramycin (positive control group); D: CP challenge+4 × 10 7  CFU/kg of feed B . subtilis PB6 (PB6 low-dosage group); and E: CP challenge+6 × 10 7  CFU/kg of feed B . subtilis PB6 (PB6 high-dosage group). There were 6 replicate pens per treatment with 15 broilers per pen. The present research examined the effect of Bacillus subtilis PB6 ( B . subtilis PB6) on growth performance, mRNA expression of intestinal cytokines and tight junctions, and gut flora composition in broilers challenged with CP. The entire experiment was divided into two phases: the non-CP challenge phase (d0-18) and the CP challenge phase (d18-26). PB6 did not increase the growth performance during the first stage, but the PB6 high-dosage group was found to have larger body weight gain and ADFI during the CP challenge stage. Feed supplementation with PB6 reduced the lesion score of challenged chicks, with increased tight junction-related gene expression ( occludin and ZO - 1 ) and decreased TNF - α expression compared with CP-infected birds. A decrease in the abundance of Clostridium XI , Streptococcu s, and Staphylococcus was observed after CP infection ( P < 0.05), while supplementation with PB6 restored the ileal microbial composition. In conclusion, administration of B . subtilis PB6 improved growth performance, enhanced intestinal barrier function, and mitigated intestinal inflammation/lesions, which might be due to its restoring effects on the ileal microbial composition in CP-challenged broilers., Competing Interests: The authors declare that they have no conflicts of interest., (Copyright © 2021 Yan Liu et al.)

Changes in functionality and composition of gut microbiota (GM) have been associated and may contribute to the development and maintenance of obesity and related diseases. The aim of our study was to investigate for the first time the impact of Lactiplantibacillus (L.) plantarum IMC 510 in a rat model of diet-induced obesity, specifically in the cafeteria (CAF) diet. This diet provides a strong motivation to voluntary overeat, due to the palatability and variety of selected energy-dense foods. The oral administration for 84 days of this probiotic strain, added to the CAF diet, decreased food intake and body weight gain. Accordingly, it ameliorated body mass index, liver and white adipose tissue weight, hepatic lipid accumulation, adipocyte size, serum parameters, including glycemia and low-density lipoprotein levels, in CAF fed rats, potentially through leptin control. In this scenario, L. plantarum IMC 510 showed also beneficial effects on GM, limiting the microbial imbalance established by long exposure to CAF diet and preserving the proportion of different bacterial taxa. Further research is necessary to better elucidate the relationship between GM and overweight and then the mechanism of action by which L. plantarum IMC 510 modifies weight. However, these promising results prompt a clear advantage of probiotic supplementation and identify a new potential probiotic as a novel and safe therapeutic approach in obesity prevention and management.

Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Mycotoxins are key risk factors in human food and animal feed. Most of food-origin mycotoxins could easily enter the organism and evoke systemic toxic effects, such as aflatoxin B1 (AFB1), ochratoxin A (OTA), T-2 toxin, deoxynivalenol (DON), zearalenone (ZEN), fumonisin B1 (FB1), and 3-nitropropionic acid (3-NPA). For the last decade, the researches have provided much evidences in vivo and in vitro that the brain is an important target organ on mycotoxin-mediated neurotoxic phenomenon and neurodegenerative diseases. As is known to all, glial cells are the best regulator and defender of neurons, and a few evaluations about the effects of mycotoxins on glial cells such as astrocytes or microglia have been conducted. The fact that mycotoxin contamination may be a key factor in neurotoxicity and glial dysfunction is exactly the reason why we reviewed the activation, oxidative stress, and mitochondrial function changes of glial cells under mycotoxin infection and summarized the mycotoxin-mediated glial cell proliferation disorders, death pathways, and inflammatory responses. The purpose of this paper is to analyze various pathways in which common food-derived mycotoxins can induce glial toxicity and provide a novel perspective for future research on the neurodegenerative diseases., Competing Interests: The authors declare that there is no conflict of interest regarding the publication of this paper., (Copyright © 2021 Xingyao Pei et al.)

This study investigated the effect of moderate risk level (8 µg/kg) AFB 1 in diet supplemented with or without adsorbents on lactation performance, serum parameters, milk AFM 1 content of healthy lactating cows and the AFM 1 residue exposure risk in different human age groups. Forty late healthy lactating Holstein cows (270 ± 22 d in milk; daily milk yield 21 ± 3.1 kg/d) were randomly assigned to four treatments: control diet without AFB 1 and adsorbents (CON), CON with 8 μg/kg AFB 1 (dry matter basis, AF), AF + 15 g/d adsorbent 1 (AD1), AF + 15 g/d adsorbent 2 (AD2). The experiment lasted for 19 days, including an AFB 1 -challenge phase (day 1 to 14) and an AFB 1 -withdraw phase (day 15 to 19). Results showed that both AFB 1 and adsorbents treatments had no significant effects on the DMI, milk yield, 3.5% FCM yield, milk components and serum parameters. Compared with the AF, AD1 and AD2 had significantly lower milk AFM 1 concentrations (93 ng/L vs. 46 ng/L vs. 51 ng/L) and transfer rates of dietary AFB 1 into milk AFM 1 (1.16% vs. 0.57% vs. 0.63%) ( p < 0.05). Children aged 2-4 years old had the highest exposure risk to AFM 1 in milk in AF, with an EDI of 1.02 ng/kg bw/day and a HI of 5.11 (HI > 1 indicates a potential risk for liver cancer). Both AD1 and AD2 had obviously reductions in EDI and HI for all population groups, whereas, the EDI (≥0.25 ng/kg bw/day) and HI (≥1.23) of children aged 2-11 years old were still higher than the suggested tolerable daily intake (TDI) of 0.20 ng/kg bw/day and 1.00 (HI). In conclusion, moderate risk level AFB 1 in the diet of healthy lactating cows could cause a public health hazard and adding adsorbents in the dairy diet is an effective measure to remit AFM 1 residue in milk and its exposure risk for humans.

The poultry industry in sub-Saharan Africa (SSA) is faced with feed insecurity, associated with high cost of feeds, and feed safety, associated with locally produced feeds often contaminated with mycotoxins. Mycotoxins, including aflatoxins (AFs), fumonisins (FBs), trichothecenes, and zearalenone (ZEN), are common contaminants of poultry feeds and feed ingredients from SSA. These mycotoxins cause deleterious effects on the health and productivity of chickens and can also be present in poultry food products, thereby posing a health hazard to human consumers of these products. This review summarizes studies of major mycotoxins in poultry feeds, feed ingredients, and poultry food products from SSA as well as aflatoxicosis outbreaks. Additionally reviewed are the worldwide regulation of mycotoxins in poultry feeds, the impact of major mycotoxins in the production of chickens, and the postharvest use of mycotoxin detoxifiers. In most studies, AFs are most commonly quantified, and levels above the European Union regulatory limits of 20 μg/kg are reported. Trichothecenes, FBs, ZEN, and OTA are also reported but are less frequently analyzed. Co-occurrences of mycotoxins, especially AFs and FBs, are reported in some studies. The effects of AFs on chickens' health and productivity, carryover to their products, as well as use of mycotoxin binders are reported in few studies conducted in SSA. More research should therefore be conducted in SSA to evaluate occurrences, toxicological effects, and mitigation strategies to prevent the toxic effects of mycotoxins.