Plastics and, in particular, microplastics (MPs) (< 5 mm) are emerging environmental pollutants responsible for interconnected risks to environmental, human, and animal health. The livestock sector is highly affected by these contaminants, with 50-60 % of the foreign bodies found in slaughtered domestic cattle being recognized as plastic-based materials. Additionally, microplastics were recently detected inside ruminant bodies and in their feces. MPs presence in ruminants could be explained by the intensive usage of plastic materials on farms, in particular to store feeds (i.e. to cover horizontal silos and to wrap hay bales). Although feed could be one of the main sources of plastics, especially of microplastics, a specific protocol to detect them in ruminant feeds is not actually present. Hence, the aim of this study was to optimize a specific protocol for the extraction, quantification, and identification of five microplastic polymers (high-density polyethylene, low-density polyethylene, polyamide fibers/particles, polyethylene terephthalate and polystyrene) from feeds typically used in ruminant diets (corn silage, hay, high protein feedstuff and total mixed ration). Several combinations of Fenton reactions and KOH digestion were tested. The final extraction protocol involved a KOH digestion (60 °C for 24 h), followed by two/three cycles of Fenton reactions. The extraction recoveries were of 100 % for high-density, low-density polyethylene, polyamide particles, and polystyrene and higher than 85 % for polyethylene terephthalate and polyamide fibers. Finally, the optimized protocol was successfully applied in the extraction of microplastics from real feed samples. All the feeds contained microplastics, particularly polyethylene, thus confirming the exposure of ruminants to MPs.

Mycotoxins are toxic secondary metabolites produced by fungal species that can cause acute, subacute, and chronic toxicity in humans and animals. Thus, these toxins pose a significant threat to health and safety. Owing to the lack of effective antimold measures in the agricultural industry, feed ingredients such as corn, peanuts, wheat, barley, millet, nuts, oily feed, forage, and their byproducts are prone to mold and mycotoxin contamination, which can affect animal production, product quality, and safety. Cyclopiazonic acid (CPA), which is mainly biosynthesized from mevalonate, tryptophan, and diacetate units, is a myotoxic secondary metabolite produced by Penicillium and Aspergillus fungi. CPA is widely present as a copollutant with aflatoxins in various crops. Compared with some common mycotoxins such as aflatoxins, fumonisins, ochratoxins, zearalenones, and their metabolites, CPA has not been well investigated. In the United States, a survey showed that 51% of corn and 90% of peanut samples contained CPA, with a maximum level of 2.9 mg/kg. In Europe, CPA was found in Penicillium-contaminated cheeses as high as 4.0 mg/kg. Some studies have shown that CPA can cause irreversible damage to organs such as the liver and spleen in mice. Therefore, the establishment of a rapid and efficient analytical method for CPA is of great significance for the risk assessment of CPA in feeds, the development of standard limits, and the protection of feed product quality and safety. The QuEChERS method, a sample pretreatment method that is fast, simple, cheap, effective, and safe, is widely used in the analysis of pesticide residues in food. In this study, a modified QuEChERS method combined with ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was used to determine CPA levels in feeds. The chromatographic separation and MS detection of CPA as well as the key factors affecting the extraction efficiency of CPA, including the type of extraction solvent, type of inorganic salt, and type and dosage of adsorbent, were optimized in detail. During the optimization of the chromatographic-separation step, the acid and salt concentrations of the mobile phase affected the separation and detection of CPA. During the optimization of the QuEChERS method, the addition of a certain amount of acetic acid improved the extraction efficiency of CPA because of its acidic nature; in addition, GCB and PSA significantly adsorbed CPA from the feed extract. Under optimal conditions, the CPA in the feed sample (1.0 g) was extracted with 2 mL of water and 4 mL of acetonitrile (ACN) containing 0.5% acetic acid. After salting out with 0.4 g of NaCl and 1.6 g of MgSO4, 1 mL of the ACN supernatant was purified by dispersive solid-phase extraction using 150 mg of MgSO4 and 50 mg of C18 and analyzed by UPLC-MS/MS. The sample was separated on a Waters HSS T3 column (100 mm×2.1 mm, 1.8 μm) using 2 mmol/L ammonium acetate aqueous solution with 0.5% formic acid and ACN as the mobile phases and then analyzed by positive electrospray ionization in multiple reaction monitoring mode. CPA exhibited good linearity in the range of 2-200 ng/mL, with a high correlation coefficient (r=0.9995). The limits of detection and quantification of CPA, which were calculated as 3 and 10 times the signal-to-noise ratio, respectively, were 0.6 and 2.0 μg/kg, respectively. The average recoveries in feed samples spiked with 10, 100, and 500 μg/kg CPA ranged from 70.1% to 78.5%, with an intra-day precision of less than 5.8% and an inter-day precision of less than 7.2%, indicating the good accuracy and precision of the proposed method. Finally, the modified QuEChERS-UPLC-MS/MS method was applied to the analysis of CPA in 10 feed samples obtained from Wuhan market. The analysis results indicated that the developed method has good applicability for CPA analysis in feed samples. In summary, an improved QuEChERS method was applied to the extraction and purification of CPA from feeds for the first time; this method provides a suitable analytical method for the risk monitoring, assessment, and standard-limit setting of CPA in feed samples.

Zebrafish are one of the most used animal models in biological research and a cost-effective alternative to rodents. Despite this, nutritional requirements and standardized feeding protocols have not yet been established for this species. This is important to avoid nutritional effects on experimental outcomes, and especially when zebrafish models are used in preclinical studies, as many diseases have nutritional confounding factors. A key aspect of zebrafish nutrition is related to feed intake, the amount of feed ingested by each fish daily. With the goal of standardizing feeding protocols among the zebrafish community, this paper systematically reviews the available data from 73 studies on zebrafish feed intake, feeding regimes (levels), and diet composition. Great variability was observed regarding diet composition, especially regarding crude protein (mean 44.98 ± 9.87%) and lipid content (9.91 ± 5.40%). Interestingly, the gross energy levels of the zebrafish diets were similar across the reviewed studies (20.39 ± 2.10 kilojoules/g of feed). In most of the reviewed papers, fish received a predetermined quantity of feed (feed supplied). The authors fed the fish according to the voluntary intake and then calculated feed intake (FI) in only 17 papers. From a quantitative point of view, FI was higher than when a fixed quantity (pre-defined) of feed was supplied. Also, the literature showed that many biotic and abiotic factors may affect zebrafish FI. Finally, based on the FI data gathered from the literature, a new feeding protocol is proposed. In summary, a daily feeding rate of 9-10% of body weight is proposed for larvae, whereas these values are equal to 6-8% for juveniles and 5% for adults when a dry feed with a proper protein and energy content is used.

Two separate feeding trials were undertaken to benchmark a series of commercial diets and determine the nutrient and energy apparent digestibility coefficients of a variety of protein-based feed ingredients when fed to sobaity seabream, Sparidentex hasta. In Experiment 1, triplicate groups of fish (initial body weight: 330.5 ± 2.6 g) were fed with one of three locally available diets containing crude protein (CP) levels ranging from 44 to 46% of dry matter (DM), each with ~12% crude fat. Fish grew at around 3.2 g day-1 with a specific growth rate (SGR) of 0.7% day-1. Both the feed conversion ratio (FCR) and protein efficiency ratio (PER) were significantly better in fish fed diets, which contained the highest (46.4%) crude protein level. Overall, the data from these preliminary studies suggest that the best performance by sobaity seabream was obtained with a diet containing 46% crude protein, 20 MJ/kg, and a protein-to-energy ratio of 23 mg/kJ. In Experiment 2, fish with an initial body weight of 319 ± 7 g were held in 11 tanks and fed reference (D1) and test diets (D2-D11) for 7 days before fecal collection. This process was repeated twice in a blocking arrangement to generate three replicates. Each of the ten test diets contained 30% of a test ingredient, with the remaining 70% proportionally identical to the D1 diet. Diet apparent digestibility coefficients (ADCs) were measured, and the diet ADCs were then used to derive the protein and energy ADCs for the individual test ingredients. Ingredient protein ADC ranged between 75.5 and 93.9%, while ingredient energy ADC ranged between 66.8 and 81.2%.

Magnetic molecularly imprinted polymers (MMIPs) have fused molecular imprinting technology with magnetic separation technology, emerging as an innovative material capable of recognizing specific molecules and efficiently separating target substances. Their application to the extraction and purification of mycotoxins has great potential, due to the toxicity and economic impact of these contaminants. In this work, MMIP has been proposed as a sample treatment for the determination of main four aflatoxins (B1, B2, G1 and G2) in pig feed. The MMIP was formed through the integration of magnetic material (Fe3O4) with commercial molecularly imprinted polymers, avoiding the synthesis step and, therefore, simplifying the process. The analyses were carried out by high-performance liquid chromatography with fluorescence detection and the method was validated and limits of quantification (LOQs) between 0.09 and 0.47 ng/g were obtained, below the allowed or recommended levels by the European Union. Repeatability and intermediate precision showed relative standard deviations lower than 10% in all cases and trueness ranged from 92 to 111%. Finally, the proposed method was applied to 31 real pig feed samples, detecting aflatoxins with concentrations between 0.2 and 3.2 ng/g.

Aflatoxins are one of the major factors that affect the quality and safety of feeds. They can be transferred into livestock through contaminated feed and then onto humans via animal sources of food such as milk, meat, and eggs. The objective of this study was to detect and quantify the level of aflatoxins (B1, B2, G1, G2, and total aflatoxin) in dairy feeds, poultry (layer and broiler) feeds, and feed ingredients produced in Addis Ababa. A total of 42 feeds and feed ingredients consisting of dairy feeds (n = 5), poultry broiler feeds (n = 6), layer feeds (n = 6), and feed ingredients (n = 25) were collected from feed factories in the city and analyzed in fresh weigh basis. The aflatoxins were analyzed using high-performance liquid chromatography after clean-up with immunoaffinity columns. Aflatoxin B1 levels in feeds ranged from 51.66 to 370.51 µg/kg in dairy cattle feed, from 1.45 to 139.51 µg/kg in poultry layer feed, and from 16.49 to 148.86 µg/kg in broiler feed. Aflatoxin B1 levels in maize ranged from 2.64 to 46.74 µg/kg and in Niger seed cake from 110.93 to 438.86 µg/kg. Aflatoxin B1 levels in wheat bran, wheat middling, and soybean were below 5 µg/kg. 100% of dairy feeds, 67% of poultry layer, 67% of broiler feeds, and 24% of ingredients contained aflatoxin in levels higher than the maximum tolerable limit set by the US Food and Drug Administration and Ethiopian Standard Agency. This shows the need for strong regulatory monitoring and better feed management practices to prevent consumers of animal-source foods from significant health impacts associated with aflatoxins.

It remains unclear how preoperative nutrition fortification impacts postoperative growth trajectories and nutritional status among infants with congenital heart disease. A single center retrospective cohort study was performed to evaluate measures of growth among patients who underwent cardiac repair at 0-18 months of age for atrial septal defect, ventricular septal defect, atrioventricular septal defect, or tetralogy of Fallot. Cohorts were analyzed at 0-30 and 31-60 days post-repair as well as at 2, 5, and 10 years of age. Records of 24 patients who received fortified nutrition and 60 patients who received unfortified nutrition preoperatively were reviewed. Those with fortified nutrition had higher growth velocities in the first 30 days post-repair compared to those with unfortified nutrition: 28.4 (23.8-83.3) grams per day versus 16.7 (7.1-21.4) grams per day, p = 0.004. Weight percentile for age was higher in the unfortified group at 2, 5, and 10 years of age (p = 0.02, p = 0.045, and p = 0.01). Body mass index (BMI) percentile for age was higher in the unfortified group at 5 and 10 years of age (p = 0.045 and p = 0.02) with a trend toward higher prevalence of either overweight or obesity compared to the fortified group (p = 0.13). reoperative nutrition fortification among infants with congenital heart disease is associated with higher growth velocity in the first 30 days post-repair and lower BMI percentile for age at 10 years. Further studies are needed to evaluate the association between preoperative nutrition fortification and postoperative outcomes, nutritional status, and prevalence of obesity in adolescence and adulthood.

Accurate dosing of feed additives is often required to evaluate their effects on rumen fermentation. This can be done using soluble but nonfermentable hydroxypropyl methylcellulose (HPMC) hard capsules.
The aim of the study was to evaluate the effect of HPMC hard capsules on the extent and rate of in vitro gas production in eleven feeds.
Six high-fibre feeds and five concentrates, were weighed into syringes either directly or into HPMC capsules and incubated anaerobically in 30 mL buffered rumen fluid at 39°C. Data obtained from gas production measurements were fitted using the Gompertz model to obtain kinetic parameters for gas production.
HPMC hard capsules had no effect on the gas production of the blank sample and concentrate feeds. In contrast, high-fibre feeds weighed in HPMC showed a significant decrease (p < 0.05) in total gas production and gas produced within 24 h of incubation.
The use of HPMC hard capsules was found to be inappropriate for determining gas production kinetics because fermentation subsides at a certain point when peak fermentation is reached (at TMFR), resulting in a decrease in both total potential gas production and gas production within 24 h of incubation. This is particularly evident when high-fibre feeds are incubated.

Dioxins (PCDD/PCDF) are toxic compounds that are ubiquitous in the environment; although present in low concentrations, they are persistent and highly toxic and they bioaccumulate in food chains. Therefore, it is very important that feed is free of these types of contaminants, because otherwise they can become a source that can negatively affect animal health and the safety of food of animal origin. The aim of the study was to comprehensively assess the concentrations of dioxins and polychlorinated biphenyls (PCBs) in a variety of feed materials available on the Polish market. In addition, characteristic profiles of congeners for given categories of feeds were investigated and defined. Approximately 95 % of the 523 samples of various feed materials tested over seven years (2013-2018 and 2022) met the requirements of European Union feed law (Commission Regulation 277/2012/EU). The highest average PCDD/PCDF/dl-PCB concentrations were found in fish oils and meal and were respectively 1.17 ± 0.78 and 5.51 ± 4.51 ng WHO-TEQ/kg of feed at 12 % moisture. Median and background level concentrations of PCDD/PCDFs, dl-PCBs, PCDD/PCDF/dl-PCBs, and ndl-PCBs were significantly lower than their average concentrations for each individual feed material category. The WHO-TEQ profiles enabled the identification of three different characteristic profiles in feed materials.

The gut microbiome plays an important role in the metabolism, nutrient absorption and immunocompetency of animals. The dynamics of the microbiota can be influenced by modulatory factors that involve nutrition, environment, health, diseases, etc. Few reports have been documented regarding the effects of different feeds on the fungi microbiome of Suffolk crossed with Tibetan sheep. A total of 30 Suffolk crossed with Tibetan sheep (ST sheep) were selected for the study and randomly divided into five equal groups (n = 6): AZ, BZ, CZ, DZ and EZ. Group AZ was fed with alfalfa and oat grass, whereas group BZ was fed with mixture of concentrated feed, alfalfa and oat grass. Groups CZ, DZ and EZ were fed with concentrated feed #1, #2 and #3, respectively. All experimental animals were fed twice a day for four months, and rectum samples were collected for microbiota analysis. Results revealed that 2,781,461 raw reads and 2,333,239 clean reads were achieved in the ST sheep. When compared with the sheep of groups AZ and BZ (164), the shared amplicon sequence variants (ASVs) between AZ and CZ (109), AZ (113) and DZ (118) as well as AZ along with EZ were fewer. Conspicuous different phyla (8) and genera (56) were examined and compared with free-range sheep in AZ. Genera including Xeromyces, Kazachstania, Cordyceps, Rhodotorula, Pichia, Spor, etc. were found higher in animals in the CZ, DZ and EZ groups. The results of this study provide new insights regarding the effects of different feeds on the fungi microbiome of sheep farmed on the plateau. We concluded that the differences in feed in Suffolk crossed with Tibetan sheep altered their gut microbiota.