Animal feed is vulnerable to fungal infections, and the use of bio-preserving probiotics has received increasing attention. In contrast to Lactobacillus and Bifidobacteria spp., fewer Bacillus spp. have been recognized as antifungal probiotics. Therefore, our objective was to screen antifungal strains and provide more Bacillus candidates to bridge this gap. Here, we screened 56 bacterial strains for cyclic lipopeptide genes and conducted an antifungal assay with Aspergillus niger as a representative fungus. We found that a Bacillus strain Bacillus amyloliquefaciens PM415, isolated from pigeon manure, exhibited the highest fungal inhibition activity as demonstrated by the confrontation assay and morphological observation under scanning electron microscope (SEM). Preliminary safety assessment and probiotic characterization revealed its non-pathogenic feature and stress tolerance capability. Whole genome sequencing of Bacillus amyloliquefaciens PM415 revealed a genome size of 4.16 Mbp and 84 housekeeping genes thereof were used for phylogenetic analysis showing that it is most closely related to Bacillus amyloliquefaciens LFB112. The in silico analysis further supported its non-pathogenic feature at the genomic level and revealed potential biosynthetic gene clusters responsible for its antifungal property. RNA-seq analysis revealed genome-wide changes in transportation, amino acid metabolism, non-ribosomal peptides (NRPs) biosynthesis and glycan degradation during fungal antagonism. Our results suggest that Bacillus amyloliquefaciens PM415 is a safe and effective probiotic strain that can prevent fungal growth in animal feeds.

Mycotoxins pose a major problem to poultry production as a result of feed contamination which has deleterious consequences such as production losses and human health risks. A total of 158 chicken feed samples were randomly collected from 46 consenting poultry farms in Oyo State throughout the wet season (April-October; 91 samples) and the dry season (November-March; 67 samples), including compounded feed (n = 129) and feed ingredients (n = 29). Samples were promptly transported to the laboratory in sterile plastic vials for lateral flow assay for mycotoxins using six different commercial mycotoxin test kits each for aflatoxin B1, zearalenone, deoxynivalenol, ochratoxin A, fumonisin, and T-2 toxin/HT-2 toxin. Summary values of mycotoxin levels (µg/kg) in the feedstuffs were represented as frequency or median (and range). Fisher exact or Mann-Whitney U tests were carried out where appropriate at α = 0.05. Every sample contained at least four mycotoxins. Aflatoxins and fumonisin co-occurred in 80% of the samples. Aflatoxin and fumonisin concentrations were above the permissible limits in 32.9% and 18.4% respectively in feedstuff sampled in the dry season while the values were 17.1% and 6.3% respectively during the wet season. Among feed ingredients, peanut cake and maize had the highest median concentration of aflatoxin and fumonisin, respectively. Median aflatoxin concentration in the feedstuff was significantly higher than the permissible limit irrespective of season. There is a need to frequently monitor mycotoxin levels of feed and feed ingredients and improve storage system for feed ingredients in order to reduce the risk associated with high mycotoxin intake in poultry.

There is a limited research focus on evaluating the detrimental effects of prolonged zearalenone (ZEN) intake on dairy cows\' health under controlled conditions. This experiment was conducted to evaluate whether the length of exposure to a ZEN-contaminated total mixed ration (TMR) at a level of 9.45 mg per day can negatively influence animal health parameters, such as milk composition, rumen and fecal fermentation, and the chewing activity of lactating dairy cows. For this experiment, we used 18 lactating Simmental cows that were fed a diet of 60% forage and 40% concentrate (on dry matter basis) for 26 consecutive days. The first 4 days were for adaptation prior to the first sampling day (day 0). The sampling events took place on day 0 (baseline) without ZEN, followed by day 1, day 7, day 14, and day 21 (with toxin). Dry matter intake (DMI) and ruminating chews per minute increased on the third week of ZEN inclusion; meanwhile, ruminating, eating, and drinking times were not affected. Most milk composition variables were also unaffected. Rumen fluid osmolality increased on day 21 and total short-chain fatty acids (SCFA) of ruminal fluid decreased on day 7. Fecal SCFA increased on day 21 and the acetate-to-propionate ratio increased from day 1 onwards, showing the influence of toxin intake. Animal health parameters, like heart rate, respiratory rate, and body temperature, were negatively influenced by ZEN intake, all increasing consistently on days 4 and 6, 9 and 12, and 16 and 18, respectively. The liver enzyme glutamate dehydrogenase decreased in response to ZEN intake on day 7. A total daily ZEN intake at the level of 9.45 mg did not show detrimental effects on DMI. Nevertheless, certain health parameters were negatively affected, including body temperature, respiratory rate, and heart rate, starting from the 7th day of ZEN intake, with additional signs of possible loss of water balance on the last sampling day.

UNASSIGNED: Bacteriophage (BP) cocktail was partially resistant to different temperatures and pH values.The BP cocktail showed lytic effects on different Salmonella isolates.The BP cocktail reduced Salmonella colonization in the internal organs of broilers.

Aflatoxin contamination in feed is a common problem in broiler chickens. The present systematic review and meta-analysis examined the impact of aflatoxin-contaminated feed and the efficacy of various feed additives on the production performance of broiler chickens fed aflatoxin-contaminated feed (AF-feed). A total of 35 studies comprising 53 AF-feed experiments were selected following PRISMA guidelines. Feed additives included in the analyses were toxins binder (TB), mannan-oligosaccharides (MOS), organic acid (OA), probiotics (PRO), protein supplementation (PROT), phytobiotics (PHY), and additive mixture (MIX). Random effects model and a frequentist network meta-analysis (NMA) were performed to rank the efficacy of feed additives, reported as standardized means difference (SMD) at 95% confidence intervals (95% CI). Overall, broiler chickens fed AF-feed had significantly lower final body weight (BW) (SMD = 198; 95% CI = 198 to 238) and higher feed conversion ratio (SMD = 0.17; 95% CI = 0.13 to 0.21) than control. Treatments with TB, MOS, and PHY improved the BW of birds fed AF-feed (P < 0.05) to be comparable with non-contaminated feed or control. Predictions on final BW from the broiler-fed aflatoxin-contaminated diet were 15% lower than the control diet. Including feed additives in the aflatoxins diet could ameliorate the depressive effect. Remarkably, our network meta-analysis highlighted that TB was the highest-performing additive (P-score = 0.797) to remedy aflatoxicosis. Altogether, several additives, especially TB, are promising to ameliorate aflatoxicosis in broiler chickens, although the efficacy was low regarding the severity of the aflatoxicosis.

The presence of insecticides like pyrethrins and synthetic pyrethroids, combined with the synergist piperonyl butoxide, in animal feeds can pose a risk to both animal and human health by contaminating the food chain. In this study, a simple and fast method was developed for the simultaneous determination of these compounds in contaminated animal feeds using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Sample preparation was carried out using a QuEChERS-based approach, and the method was validated with acceptable accuracy ranging from 84 to 115% and precision below 10%. The limit of detection (LOD) and limit of quantification (LOQ) were between 0.15 and 3 and 1 and 10 µg/kg, respectively. The method detected insecticide contaminations in various livestock and poultry feeds. Furthermore, the method was applied to a toxicology case, where it identified and quantified piperonyl butoxide and deltamethrin in the submitted horse feed sample. These results demonstrate that the method can be a valuable tool in animal health and food safety diagnostic applications, as well as veterinary toxicology investigations concerning pyrethrin-related feed contamination.

Camel milk is known as a source of nutritional and health supplements. It is known to be rich in peptides and functional proteins. One main issue facing it is related to its contamination, mainly with aflatoxins. The present study aimed to evaluate camel milk samples from different regions while trying to reduce its toxicity using safe approaches based on probiotic bacteria. Collected samples of camel milk were sourced from two main regions: the Arabic peninsula and North Africa. Samples were tested for their contents of aflatoxins (B1 and M1) using two techniques to ensure desired contamination levels. Additionally, feed materials used in camel foods were evaluated. Applied techniques were also tested for their validation. The antioxidant activity of camel milk samples was determined through total phenolic content and antioxidant activity assays. Two strains of probiotic bacteria (Lactobacillus acidophilus NRC06 and Lactobacillus plantarum NRC21) were investigated for their activity against toxigenic fungi. The result revealed high contamination of aflatoxin M1 for all samples investigated. Furthermore, cross-contamination with aflatoxin B1 was recorded. Investigated bacteria were recorded according to their significant inhibition zones against fungal growth (11 to 40 mm). The antagonistic impacts were between 40% and 70% against toxigenic fungi. Anti-aflatoxigenic properties of bacterial strains in liquid media were recorded according to mycelia inhibition levels between 41 to 52.83% against Aspergillus parasiticus ITEM11 with an ability to reduce aflatoxin production between 84.39% ± 2.59 and 90.4% ± 1.32 from media. Bacteria removed aflatoxins from the spiked camel milk in cases involving individual toxin contamination.

Food waste and feed-food competition can be reduced by replacing traditional feed ingredients such as cereals, with former food products (FFPs) in livestock diets. These foodstuffs, initially intended for human consumption, are recovered, mechanically unpacked, and then ground. Despite this simple and inexpensive treatment, packaging contaminants (remnants) are often unavoidable in the final product. To maximize the exploitation of FFPs and to minimize the associated risks, packaging remnants need to be quantified and characterized. This study tested the efficacy of the Fourier Transform Infrared Spectroscopy coupled with an optical microscope (μFT-IR) in identifying packaging remnants in 17 FFP samples collected in different geographical areas. After a visual sorting procedure, presumed packaging remnants were analyzed by μFT-IR. The results showed significant differences (p < 0.05) between the FFPs in terms of the total number of foreign particles found (plastics, cellulose and aluminum remnants, ranging from 4 to 19 particles per 20 g fresh matter), and also regarding the number of cellulose and aluminum particles. These data clearly demonstrate the need for sensitive instruments that can characterize the potential contaminants in the FFPs. This would then help to reduce the overestimation of undesirable contaminants typical of simple visual sorting, which is currently the most common method.

The first feed offered to young chicks is likely the most important meal in their life. The complex gut colonisation process is determined with early exposure and during the first days of life before the microbial community is formed. Therefore, providing access to high-quality feed and an environment enriched in the beneficial and deprived of pathogenic microorganisms during this period is critical. Feed often carries a complex microbial community that can contain major poultry pathogens and a range of chemical contaminants such as heavy metals, mycotoxins, pesticides and herbicides, which, although present in minute amounts, can have a profound effect on the development of the microbial community and have a permanent effect on bird\'s overall health and performance. The magnitude of their interference with gut colonisation in livestock is yet to be determined. Here, we present the animal feed quality issues that can significantly influence the microbial community development, thus severely affecting the bird\'s health and performance.

BACKGROUND: The effect in a sialochemistry profile of the presence of usually available feed in dairy cows was evaluated by an in vitro experiment. For this purpose, a pooled clean saliva from five healthy dairy cows was incubated five times with a standard feed based on a total mixed ration (F), wheat hay (H), and grass (G). The salivary panel was integrated by biomarkers of stress (cortisol -sCor-, salivary alpha-amylase -sAA-, butyrylcholinesterase -BChE-, total esterase -TEA-, and lipase -Lip-), immunity (adenosine deaminase -ADA-), oxidative status (Trolox equivalent antioxidant capacity -TEAC-, the ferric reducing ability of saliva -FRAS-, the cupric reducing antioxidant capacity -CUPRAC-, uric acid, and advanced oxidation protein products -AOPP-), and enzymes, proteins, and minerals of general metabolism and markers of liver, muscle, and renal damage (aspartate aminotransferase -AST-, alanine aminotransferase -ALP-, γ-glutamyl transferase -gGT-, lactate dehydrogenase -LDH-, creatine kinase -CK-, creatinine, urea, triglycerides, glucose, lactate, total protein, phosphorus, and total calcium).
RESULTS: Most of the evaluated analytes showed a coefficient of variations (CV) higher than 15% and/or significant changes compared with the clean saliva when feed was present. Some analytes, such as the oxidative status biomarkers (CV > 80%), AST (CV > 60%), or glucose (CV > 100%), showed significant changes with all the feed types tested. Others showed significant differences only with certain types of feed, such as LDH with F (CV > 60%) or triglycerides with F (CV > 100%) and H (CV > 95%). However, sCor or gGT remained unchanged (CV < 15%, P > 0.05) in all the treatments.
CONCLUSIONS: The presence of feed can produce changes in most of the analytes measured in cows\' saliva, being of high importance to consider this factor when saliva is used as a sample to avoid errors in the interpretation of the results.