L. Pinotti

Comparative mammalian choline metabolism with emphasis on the high-yielding dairy cow.

The present review examines the importance of choline in dairy cow nutrition. Choline is an essential nutrient for mammals when excess methionine and folate are not available in the diet. The requirement for choline can be met by dietary choline and by transmethylation reactions. Two types of functions for choline are known: functions of choline per se; functions as a methyl donor. The two principal methyl donors in animal metabolism are betaine, a metabolite of choline, and S-adenosyl-methionine, a metabolite of methionine. Choline and methionine are interchangeable with regard to their methyl group-furnishing functions. In adult ruminants, choline is extensively degraded in the rumen; for this reason dietary choline contributes insignificantly to the choline body pool and methyl group metabolism is generally conservative with a relatively low rate of methyl catabolism and an elevated rate of de novo synthesis of methyl groups via the tetrahydrofolate system. In dairy ruminants, the dietary availability of choline is still low, but the output of methylated compounds in milk is high, and precursors from the tetrahydrofolate pathway are limiting, especially at the onset of lactation. Therefore choline may be a limiting nutrient for milk production in high-yielding dairy cows.

Mycotoxin Contamination in the EU Feed Supply Chain: A Focus on Cereal Byproducts

Mycotoxins represent a risk to the feed supply chain with an impact on economies and international trade. A high percentage of feed samples have been reported to be contaminated with more than one mycotoxin. In most cases, the concentrations were low enough to ensure compliance with the European Union (EU) guidance values or maximum admitted levels. However, mycotoxin co-contamination might still exert adverse effects on animals due to additive/synergistic interactions. Studies on the fate of mycotoxins during cereal processing, such as milling, production of ethanol fuels, and beer brewing, have shown that mycotoxins are concentrated into fractions that are commonly used as animal feed. Published data show a high variability in mycotoxin repartitioning, mainly due to the type of mycotoxins, the level and extent of fungal contamination, and a failure to understand the complexity of food processing technologies. Precise knowledge of mycotoxin repartitioning during technological processes is critical and may provide a sound technical basis for feed managers to conform to legislation requirements and reduce the risk of severe adverse market and trade repercussions. Regular, economical and straightforward feed testing is critical to reach a quick and accurate diagnosis of feed quality. The use of rapid methods represents a future challenge.

Use of the electronic nose as a screening tool for the recognition of durum wheat naturally contaminated by deoxynivalenol: a preliminary approach.

Fungal contamination and the presence of related toxins is a widespread problem. Mycotoxin contamination has prompted many countries to establish appropriate tolerance levels. For instance, with the Commission Regulation (EC) N. 1881/2006, the European Commission fixed the limits for the main mycotoxins (and other contaminants) in food. Although valid analytical methods are being developed for regulatory purposes, a need exists for alternative screening methods that can detect mould and mycotoxin contamination of cereal grains with high sample throughput. In this study, a commercial electronic nose (EN) equipped with metal-oxide-semiconductor (MOS) sensors was used in combination with a trap and the thermal desorption technique, with the adoption of Tenax TA as an adsorbent material to discriminate between durum wheat whole-grain samples naturally contaminated with deoxynivalenol (DON) and non-contaminated samples. Each wheat sample was analysed with the EN at four different desorption temperatures (i.e., 180 °C, 200 °C, 220 °C, and 240 °C) and without a desorption pre-treatment. A 20-sample and a 122-sample dataset were processed by means of principal component analysis (PCA) and classified via classification and regression trees (CART). Results, validated with two different methods, showed that it was possible to classify wheat samples into three clusters based on the DON content proposed by the European legislation: (a) non-contaminated; (b) contaminated below the limit (DON < 1,750 μg/kg); (c) contaminated above the limit (DON > 1,750 μg/kg), with a classification error rate in prediction of 0% (for the 20-sample dataset) and 3.28% (for the 122-sample dataset).

Sampling feed for mycotoxins: acquiring knowledge from food

Abstract The occurrence and control of mycotoxins in feed and food are items of great interest to researchers, producers, manufacturers and regulatory agencies. In order to implement knowledge of control measures for mycotoxins in the entire food production chain, coordinated inspection programmes aimed to check the presence and concentration of mycotoxins in feedingstuffs are recommended by the Commission of the European Communities. Reliability of measured levels of mycotoxins in feed and food is greatly affected by the collection of representative samples. Because of the heterogeneous distribution of mycotoxins, the variability associated with a mycotoxin test procedure usually depends heavily on the sampling plan. European legislation dealing with sampling plans for mycotoxins in foodstuffs has been recently revised. The aim of the following overview is to discuss the role of sampling in mycotoxin-contaminated feed by considering the evolution of legislation dealing with sampling plans for food. A sampling procedure is a multistage process and consists of three distinct phases: sampling, sample preparation and analysis. The variability associated with each step of a sampling procedure and the aspects related to feedstuffs, matrix/mycotoxin combination and level of contamination are discussed.

Alpha-Tocopherol Counteracts the Cytotoxicity Induced by Ochratoxin A in Primary Porcine Fibroblasts

The aims of the current study were to determine the half-lethal concentration of ochratoxin A (OTA) as well as the levels of lactate dehydrogenase release and DNA fragmentation induced by OTA in primary porcine fibroblasts, and to examine the role of α-tocopherol in counteracting its toxicity. Cells showed a dose-, time- and origin-dependent (ear vs. embryo) sensitivity to ochratoxin A. Pre-incubation for 3 h with 1 nM α-tocopherol significantly (P < 0.01) reduced OTA cytotoxicity, lactate dehydrogenase release and DNA damage in both fibroblast cultures. These findings indicate that α-tocopherol supplementation may counteract short-term OTA toxicity, supporting its defensive role in the cell membrane.

Implementation of the electronic nose for the identification of mycotoxins in durum wheat (Triticum durum)

The on-line evaluation of the quality of durum wheat (Triticum durum) represents, in terms of health and safety, one of the leading challenges of the milling industry. Knowledge of the content and distribution of mycotoxins on wheat kernel is very economically relevant and has great health implications due to its high impact on human health and on the safety of the use of decortication products in animal nutrition. The production of mycotoxins by particular mould strains is generally associated with the production of volatile substances such as alcohols, aldehydes, ketones and esters (Magan and Evans, 2000). On this basis we intended to develop the innovative method of the sensorial analysis (based on the use of an “electronic nose”), with the aim of evaluating its potential use as a rapid diagnostic test for determination of the levels of mycotoxin contamination in durum wheat.

Rumen-protected choline and vitamin E supplementation in periparturient dairy goats: effects on milk production and folate, vitamin B 12 and vitamin E status

We investigated the effects of rumen-protected choline (RPC) and vitamin E (VITE) administration on milk production and status of folate, vitamin B12 and vitamin E during the periparturient period of dairy goats. Forty-eight Saanen multiparous goats were selected for the 72-day experiment, being moved to a maternity pen 30 days before expected parturition and assigned to one of the four experimental groups: control (CTR), no choline or vitamin E supplementation; choline (RPC), supplemented with 4 g/day choline chloride in rumen-protected form; vitamin E (VITE), supplemented with 200 IU/day vitamin E in rumen-protected form; and choline and vitamin E (RPCE), supplemented with 4 g/day RPC chloride and 200 IU/day vitamin E. Supplements were administered individually before the morning feed to ensure complete consumption, starting 30 days before kidding and continuing for 35 days after. During the experiment, milk yield and 4% fat-corrected milk (FCM) yield were, respectively, 210 and 350 g/day higher in RPC-supplemented goats than in non-supplemented goats. Milk fat concentration and fat yield were also increased by RPC treatment. Milk yield and composition were unaffected by vitamin E supplementation. There were no significant interactions between RPC and VITE for any of the variables measured. Plasma metabolites did not differ between treatments before and after kidding except that plasma folate at parturition was higher in RPC-supplemented goats. Neither choline nor vitamin E affected vitamin B12 plasma concentrations, while a time effect was evident after the second week of lactation, when B12 levels in each treatment group started to increase. Vitamin E administration resulted in plasma α-tocopherol levels that were 2 to 2.5 times higher than in non-supplemented goats. Overall, these results suggest that greater choline availability can improve milk production and methyl group metabolism in transition dairy goats.

A survey of the mycobiota associated with larvae of the black soldier fly (Hermetia illucens) reared for feed production

Feed security, feed quality and issues surrounding the safety of raw materials are always of interest to all livestock farmers, feed manufacturers and competent authorities. These concerns are even more important when alternative feed ingredients, new product developments and innovative feeding trends, like insect-meals, are considered. The black soldier fly (Hermetia illucens) is considered a good candidate to be used as feed ingredient for aquaculture and other farm animals, mainly as an alternative protein source. Data on transfer of contaminants from different substrates to the insects, as well as the possible occurrence of toxin-producing fungi in the gut of non-processed insects are very limited. Accordingly, we investigated the impact of the substrate/diet on the intestinal mycobiota of H. illucens larvae using culture-dependent approaches (microbiological analyses, molecular identification through the typing of isolates and the sequencing of the 26S rRNA D1/D2 domain) and amplicon-based next-generation sequencing (454 pyrosequencing). We fed five groups of H. illucens larvae at the third growing stage on two substrates: chicken feed and/or vegetable waste, provided at different timings. The obtained results indicated that Pichia was the most abundant genus associated with the larvae fed on vegetable waste, whereas Trichosporon, Rhodotorula and Geotrichum were the most abundant genera in the larvae fed on chicken feed only. Differences in the fungal communities were highlighted, suggesting that the type of substrate selects diverse yeast and mold genera, in particular vegetable waste is associated with a greater diversity of fungal species compared to chicken feed only. A further confirmation of the significant influence of diet on the mycobiota is the fact that no operational taxonomic unit common to all groups of larvae was detected. Finally, the killer phenotype of isolated yeasts was tested, showing the inhibitory activity of just one species against sensitive strains, out of the 11 tested species.

Computer image analysis: an additional tool for the identification of processed poultry and mammal protein containing bones.

The aims of this study were (1) to evaluate the potential of image analysis measurements, in combination with the official analytical methods for the detection of constituents of animal origin in feedstuffs, to distinguish between poultry versus mammals; and (2) to identify possible markers that can be used in routine analysis. For this purpose, 14 mammal and seven poultry samples and a total of 1081 bone fragment lacunae were analysed by combining the microscopic methods with computer image analysis. The distribution of 30 different measured size and shape bone lacunae variables were studied both within and between the two zoological classes. In all cases a considerable overlap between classes meant that classification of individual lacunae was problematic, though a clear separation in the means did allow successful classification of samples on the basis of averages. The variables most useful for classification were those related to size, lacuna area for example. The approach shows considerable promise but will need further study using a larger number of samples with a wider range.

Selection of new markers for animal by-products characterization by classical microscopy

Abstract The aim of this study was to identify possible markers to distinguish differences between land animals by using the microscopic method in association with computer image analysis. For this purpose bone fragments from poultry and mammals were obtained and analysed by microscopic method. Through a digital camera and an image analysis software 85 bone lacunae images have been processed and elaborated in order to obtain for each lacuna a monochrome mask on which several measurements were performed. Data were analysed by ANOVA and LDA. Results obtained in the present study indicated that of 32 descriptors processed by image analysis software, only 12 were significantly (P<0.001) different between mammalian and poultry. However, when morphometric measurements were analysed by LDA, 86% of lacunae were correctly classified into the animal class of origin (i.e. mammalian as mammalian and poultry as poultry). By contrast 14% of lacunae were incorrectly classified. In conclusion, data here presented indicate that some of descriptors used by image analysis appears promising not only for a reliable distinction between the different origins of animal meal at the level of vertebrate classes, but also for further characterisation and identification of processed animal proteins in animal feeds.