Nutritional and nutraceutical quality of donkey milk

In the last few decades, there has been a renewed interest in donkeys by the scientific community involved in the recovery of biodiversity, in the rescue of some donkey breeds that have become almost extinct, and in the rediscovery of donkey milk. In addition, due to the increase in food allergies, attention has been focused on the need for a “natural” milk with a good taste, which could be used in some childhood illnesses such as allergy to cow's milk proteins allergy (CMPA). Donkey milk is very similar to human milk, especially in terms of its protein profile and lactose content, which increases palatability, stimulates intestinal absorption of calcium, and thus provides an excellent substrate for the development of enteric flora. The high content of lysozyme in this milk favours selective action against pathogenic microorganisms. In addition, the mineral content (such as calcium) and liposoluble vitamins make it an excellent nutraceutical product. A project aimed to create a donkey milk supply chain from the Amiata native breed has recently been developed in Central Italy (Tuscany). In this paper, the nutritional and nutraceutical characteristics of donkey milk are reviewed in detail. In addition, some of the potential uses of donkey milk have been briefly described: in the diet of children with CMPA, in the diets of the elderly and of people who need to lose weight by virtue of the low fat content and the good contribution of omega 3

Social Preference Tests in Zebrafish: A Systematic Review

The use of animal models in biology research continues to be necessary for the development of new technologies and medicines, and therefore crucial for enhancing human and animal health. In this context, the need to ensure the compliance of research with the principles Replacement, Reduction and Refinement (the 3 Rs), which underpin the ethical and human approach to husbandry and experimental design, has become a central issue. The zebrafish (Danio rerio) is becoming a widely used model in the field of behavioral neuroscience. In particular, studying zebrafish social preference, by observing how an individual fish interacts with conspecifics, may offer insights into several neuropsychiatric and neurodevelopmental disorders. The main aim of this review is to summarize principal factors affecting zebrafish behavior during social preference tests. We identified three categories of social research using zebrafish: studies carried out in untreated wild-type zebrafish, in pharmacologically treated wild-type zebrafish, and in genetically engineered fish. We suggest guidelines for standardizing social preference testing in the zebrafish model. The main advances gleaned from zebrafish social behavior testing are discussed, together with the relevance of this method to scientific research, including the study of behavioral disorders in humans. The authors stress the importance of adopting an ethical approach that considers the welfare of animals involved in experimental procedures. Ensuring a high standard of animal welfare is not only good for the animals, but also enhances the quality of our science

Management Factors Influence AnimalWelfare and the Correlation to Infectious Diseases in Dairy Cows

Simple Summary: To investigate the relationship between some infectious diseases (Mycobacterium avium subsp. paratuberculosis, Chlamydiophila abortus, Neospora caninum, bovine viral diarrhea virus, and the bovine herpesvirus) and the dairy farms’ welfare scores, 36 dairy farms were monitored using the Italian National AnimalWelfare Reference Center (CreNBA) checklist. Farms and their animals were scored in five different areas, namely: Area A, “Farm management and personnel”; Area B, “Facilities and equipment”; Area C, “Animal-based measures”; Area D, “Inspection of microclimatic environmental conditions and alarm systems”; and Area E, “Biosecurity”. The recorded scores were compared between two farming conditions (access to pasture and indoor housing) and correlated with the serum data. Our results indicated that an accurate application of the checklist could be an instrument to prevent and control the spread of infections in dairy farms. Abstract: The present study assessed dairy cow welfare through the application of the Italian National Animal Welfare Reference Center (CReNBA) checklist in 36 dairy farms located in Ragusa (Italy) subjected to two different management conditions, housing with free access to pasture (Group 1, farms n = 17) and indoor housing (Group 2, farms n = 19). Five areas of investigation were considered: Area A, “Farm management and personnel”; Area B, “Facilities and equipment”; Area C, “Animal-based measures”; Area D, “Inspection of microclimatic environmental conditions and alarm systems”; and Area E, “Biosecurity”. Blood samples were collected by coccygeal venipuncture from all animals (4081 cows). The specific antibodies against Mycobacterium avium subsp. paratuberculosis, Chlamydiophila abortus, Neospora caninum, bovine viral diarrhea virus, and the bovine herpesvirus were assessed by enzyme linked immunosorbent assay (ELISA) serological test. Group 1 (access to pasture) showed a lower value of percentage score recorded in Area A (p = 0.02) and E (p = 0.01) than Group 2 (indoor housing). Herpesvirus (Infectious bovine rhinotracheitis - IBR - detection of gB antibodies/IBR-gB) blood concentrations were higher in the cows housed indoor versus those with access to pasture (p = 0.01). Farm management and personnel (score A) was correlated with the level of bovine viral diarrhea virus ( = 0.3754) and bovine-herpesvirus-specific antibodies (IBR-gB) ( = 0.4159). “Biosecurity” percentage score showed a significant correlation with Chlamydiophila abortus ( = 0.4621) in the cows with access to pasture and IBR-gB ( = 0.3435) in the cows housed fully indoors. Group 2 showed a significantly reduced level of antibodies against Neospora caninum. In conclusion, differences in the welfare assessment score were observed in the “Farm management and personnel” and “Biosecurity” between the two management conditions. It had an effect on the prevalence of herpesvirus, which occurred more in cattle with access to pasture. Therefore, an accurate application of the checklist could be an instrument to prevent and control the spread of infections in farms

Short-Term Effects of Human versus Bovine Sialylated Milk Oligosaccharide Microinjection on Zebrafish Larvae Survival, Locomotor Behavior and Gene Expression

Milk oligosaccharides are a complex class of carbohydrates that act as bioactive factors in numerous defensive and physiological functions, including brain development. Early nutrition can modulate nervous system development and can lead to epigenetic imprinting. We attempted to increase the sialylated oligosaccharide content of zebrafish yolk reserves, with the aim of evaluating any short-term effects of the treatment on mortality, locomotor behavior, and gene expression. Wild-type embryos were microinjected with saline solution or solutions containing sialylated milk oligosaccharides extracted from human and bovine milk. The results suggest that burst activity and larval survival rates were unaffected by the treatments. Locomotion parameters were found to be similar during the light phase between control and treated larvae; in the dark, however, milk oligosaccharide-treated larvae showed increased test plate exploration. Thigmotaxis results did not reveal significant differences in either the light or the dark conditions. The RNA-seq analysis indicated that both treatments exert an antioxidant effect in developing fish. Moreover, sialylated human milk oligosaccharides seemed to increase the expression of genes related to cell cycle control and chromosomal replication, while bovine-derived oligosaccharides caused an increase in the expression of genes involved in synaptogenesis and neuronal signaling. These data shed some light on this poorly explored research field, showing that both human and bovine oligosaccharides support brain proliferation and maturation

Chemical genetics strategies for identification of molecular targets

Chemical genetics is an emerging field that can be used to study the interactions of chemical compounds, including natural products, with proteins. Usually, the identification of molecular targets is the starting point for studying a drug’s mechanism of action and this has been a crucial step in understanding many biological processes. While a great variety of target identification methods have been developed over the last several years, there are still many bioactive compounds whose target proteins have not yet been revealed because no routine protocols can be adopted. This review contains information concerning the most relevant principles of chemical genetics with special emphasis on the different genomic and proteomic approaches used in forward chemical genetics to identify the molecular targets of the bioactive compounds, the advantages and disadvantages of each and a detailed list of successful examples of molecular targets identified with these approaches

Studio sull'impiego di farina di insetto in sostituzione della farina di pesce, utilizzando lo zebrafish come modello animale

La farina di pesce è un importante ingrediente dei mangimi destinati alle specie ittiche carnivore. Purtroppo, a causa della riduzione degli stock ittici selvatici, negli ultimi decenni è stato osservato un drastico aumento del suo prezzo di mercato. Di conseguenza, l’industria mangimistica e la comunità scientifica, stanno cercando valide alternative e ad oggi a tale scopo vengono principalmente impiegate materie prime di origine vegetale (soia, estratti di soia, glutine di mais e altre granelle), caratterizzate da profili amminoacidici non adeguati, dalla presenza di fattori anti-nutrizionali e dalla ridotta digeribilità. In tale contesto, la farina di insetto può rappresentare una valida fonte proteica alternativa ed il basso “carbon footprint” la rende interessante anche nell'ottica di migliorare la “sostenibilità delle produzioni”. L’obiettivo di questo studio è stato quello di valutare l’effetto della farina di insetto, in sostituzione alla farina di pesce, nelle diete per le specie ittiche. La farina di insetto utilizzata è stata prodotta con larve di Hermetia illucens (Insect protein meal®, Protix©, Dongen - NL). Lo studio è stato condotto presso la “zebrafish facility” dello stabulario centralizzato situato presso il Dipartimento di Scienze Veterinarie dell’Università di Pisa, utilizzando lo zebrafish (Danio rerio) come modello animale. A questo scopo, 320 giovanili di zebrafish (Wild-Type; Linea AB; peso vivo 85±25 mg (m±ds); 60 giorni di età;), suddivisi in maniera casuale in 4 gruppi e 4 repliche, sono stati alimentati con diete che differivano per il rapporto di sostituzione della farina di pesce con la farina di insetto; in particolare: 0% (gruppo controllo alimentato con il 20% di farina di pesce), 25% (HI25), 50% (HI50) e 100% (HI100). Le diete sperimentali erano iso-energetiche (21.8±0.2 MJ kg−1 di s.s.), iso-proteiche (60.1±0.3% proteina grezza/s.s.) ed estruse. Queste diete sono state prodotte dalla Sparos© (Olhão - P), un’azienda spin-off dell’Università di Algarve. I pesci sono stati alimentati 4 volte al giorno per un periodo di 49 giorni. L’effetto delle diete sui pesci è stato investigato mediante le performance di accrescimento, la mortalità, il consumo di mangime e l’indice di conversione alimentare. Al termine della prova gli zebrafish hanno quasi triplicato il loro peso vivo. Per tutti i parametri considerati, non sono state osservate differenze significative (p>0.05) sia tra i gruppi sperimentali, che tra questi ed il gruppo di controllo; il peso vivo medio finale raggiunto dai ogni gruppo è stato pari a: controllo 229 mg; HI25 237 mg; HI50 242 mg; HI100 229 mg, mentre l’indice di conversione alimentare è stato pari a: controllo 2.11; HI25 2.05; HI50 1.94; HI100 2.08. Le uniche differenze statisticamente significative (p<0.05) sono state osservate tra maschi e femmine, quindi indipendentemente dal trattamento. In conclusione, i risultati suggeriscono che la farina di larve di Hermetia illucens rappresenta una valida alternativa alla farina di pesce nei mangimi per le specie ittiche allevate, anche se ulteriori studi dovranno essere condotti su altre specie ittiche

Studio di alcuni fattori che influenzano le performance produttive dell'asina: management degli animali, componenti nutraceutici del latte ed applicazioni pratiche del prodotto

Per millenni, la specie asinina è stata utilizzata dall’uomo per la sua natura paziente come animale da lavoro e per la sua rusticità. Dall’inizio della sua domesticazione comunque, le asine sono state utilizzate anche per la produzione di latte, dalle leggendarie proprietà nutrizionali e terapeutiche. Negli ultimi secoli, la meccanizzazione agricola e dei trasporti ha causato una forte diminuzione della popolazione asinina mondiale, ma attualmente, il numero degli allevamenti asinini e la consistenza delle popolazioni autoctone sono in crescita a livello Europeo. Il latte d’asina viene oggi adoperato primariamente nell’alimentazione dei bambini e nella cosmesi. La ricerca scientifica sul contenuto e la variabilità di particolari nutrienti bio-attivi del latte e sulle tecniche di allevamento delle asine, è tuttavia assai limitata. L’obiettivo del lavoro è stato quello di studiare alcuni fattori che influenzano le performance produttive dell’asina da latte e ampliare le conoscenze sul contenuto e la variabilità di particolari composti nutraceutici, al fine di fornire indicazioni utili ai futuri piani di selezione della specie. Scopo del lavoro è stato anche quello di valutare dettagliatamente, la qualità nutrizionale del latte nei primi 10 giorni di lattazione, la curva di accrescimento dei puledri e le variazioni di peso vivo e ingestione alimentare delle asine in diverse condizioni fisiologiche. Infine, tramite un test in vitro è stata valutata l’attività antimicotica del latte d’asina e tramite un trial clinico è stata valutata l’allergenicità del latte d’asina nell’alimentazione dei bambini allergici alle proteine del latte vaccino. Tutte le ricerche effettuate in questo lavoro, sono state condotte su asini appartenenti ad una popolazione autoctona Toscana: l’asino dell’Amiata. Gli animali utilizzati sono stati allevati in purezza e con sistema semi-intensivo presso l’allevamento “Bandite di Scarlino”, situato in provincia di Grosseto. La composizione chimico-fisica del latte e degli alimenti è stata analizzata utilizzando i metodi dell’Assocazione Ufficiale di Chimica Analitica (AOAC, 2000) e metodi validati. L’attività di ricerca ha permesso di ampliare le conoscenze sul contenuto/attività e la variabilità dei seguenti composti nutraceutici del latte d’asina: oligosaccaridi sialici, vitamina D e lisozima. I risultati dimostrano che il latte d’asina appare essere una potenziale fonte di oligosaccaridi sialici, ed in particolare di 6’-SL (media circa 19 mg/L), e di vitamina D (media 1.68 μg/100 mL nel latte pastorizzato. Inoltre, il contenuto totale di vitamina D non diminuisce a seguito del trattamento di pastorizzazione o nelle diverse stagioni, mentre la quantità di vitamina D3 si riduce della metà a seguito del trattamento termico, e durante la stagione primaverile-estiva il suo contenuto è maggiore rispetto al periodo autunnale-invernale. In relazione al lisozima, i risultati dimostrano che l’attività di quest’enzima è maggiore nel latte delle asine a inizio lattazione e in quelle di età superiore ai 15 anni. Inoltre, il contenuto di calcio, ceneri e di alcuni acidi grassi è correlato positivamente ad una maggiore attività del lisozima a inizio lattazione. È stato analizzato inoltre il polimorfismo delle proteine del latte e quello dei geni del lisozima e dell’ossitocina, fornendo indicazioni utili su particolari genotipi capaci di produrre un latte con determinate caratteristiche nutrizionali. Infatti, i risultati di questo studio mostrano che ai diversi pattern genetici identificati, sia associata una differente composizione del profilo proteico. Tramite le tecniche di elettroforesi utilizzate, è stata quantificata per la prima volta in questo studio il contenuto di κ-caseina del latte d’asina. Sono state approfondite le conoscenze sulle caratteristiche nutrizionali e citologiche del latte d’asina e l’effetto del trattamento di pastorizzazione sulla qualità nutrizionale. In relazione alla composizione del latte nei primi 10 giorni di lattazione, per quasi tutti i parametri chimico-fisici analizzati, delle differenze significative sono state osservate. Sono state monitorate le variazioni di peso vivo, Body Condition Score, ingestione alimentare e i parametri ematologici della specie nelle diverse condizioni fisiologiche. I risultati hanno evidenziato che le asine Amiatine in mantenimento dovrebbero essere alimentate con solo fieno ad libitum (con energia digeribile leggermente superiore a 8 MJ/kg di foraggio secco), in quantità pari al 2.5% del loro peso vivo. L’incremento ponderale giornaliero medio dei puledri, nei primi 10 giorni di vita, è stato pari a 0.962 kg e a 2 mesi, i puledri si sono accresciuti del 126% rispetto al peso alla nascita. Uno studio preliminare sul miglioramento dell’efficienza nella mungitura è stato effettuato ricorrendo a 4 mungiture giornaliere. I risultati osservati suggeriscono che, all’aumentare del numero di mungiture giornaliere, sia la quantità di latte ottenibile per singola mungitura, che il tenore lipidico aumenta. Infine, un test in vitro ha dimostrato che il latte d’asina è capace di inibire la crescita di alcuni dermatofiti zoonotici (T. mentagrophytes, M. canis e M. gypseum), e un trial clinico ha evidenziato che il latte d’asina è stato tollerato dal 98.7% dei bambini arruolati nello studio, confermando l’ipoallergenicità e la buona palatabilità di questo latte nell’alimentazione umana

Technological and seasonal variations of vitamin D and other nutritional components in donkey milk

Vitamin D is an essential nutrient that plays a crucial role in calcium homeostasis and bone metabolism and also acts as a hormone. Although several studies on the content of vitamin D in bovine milk have been conducted, little information is available regarding donkey milk. In the context of the nutritional assessment of donkey milk, the aim of this study was to assess the vitamin D content in donkey milk and its chemical profile, with particular reference to seasonal and technological modifications after pasteurization. The study was conducted on a dairy farm that produces donkey milk for human consumption located in central Italy. At sampling time, an aliquot of total bulk milk production was sampled before and after pasteurization (63°C for 30 min without homogenization) with a total of 20 raw and 20 pasteurized milk samples. The samples were collected for 10 mo, every 15 d, from May to February 2017. All the samples were analyzed for the chemical composition and vitamin D2 and D3 content by HPLC after saponification. The donkey milk analyzed showed a higher average vitamin D content (raw milk: vitamin D2 = 1.68, vitamin D3 = 0.60 μg/100 mL; pasteurized milk: vitamin D2 = 1.38, vitamin D3 = 0.30 μg/100 mL) than reported for bovine and human milk. The results of the effect of pasteurization on milk did not highlight significant differences in the total content of vitamin D. However, vitamin D3 has a poor thermal stability, which led to a significant reduction in content in pasteurized milk compared with raw milk. The total vitamin D content of donkey milk did not show significant variations between seasons; however, a higher concentration of vitamin D3 was found in spring and summer. In conclusion, raw and pasteurized donkey milk showed a high content of vitamin D, which could be useful in meeting the deficiencies of this vitamin in humans. Further investigations are needed to improve the vitamin D content in donkey milk by increasing its endogenous synthesis or its transfer in milk and to clarify other variability factors. © 2018 American Dairy Science Associatio

Nutritional and nutraceutical Amiatina donkey milk quality.

Recently, a project aimed to the creation of a donkey milk chain from Amiatina native breed has been developed in Tuscany. In a second step of the project, the use of donkey milk in the diet of children with allergy to the cow's milk proteins (CMA) has been evaluated. At the same time, nutritional, nutraceutical, hygienic and safety milk characteristics were tested. Furthermore, another topic of the project was to promote the biodiversity and the economy of Tuscany productions and to provide products having a constant quality. Every month bulk milk samples underwent physical and chemical analysis. The results showed dry matter content of 10 g/100 ml of milk and total protein and fat content of 1.6 and 0.5 g/100 ml, respectively. In donkey milk the protein fraction was more similar to human milk than ruminant milk, with low amounts of casein (0.8 g/100 ml). Whereas whey protein content was about a half of the total proteins (51.1%). The main whey protein fractions were beta-lactoglobulin and alpha-lactalbumin (0.35 and 0.30 g/100 ml, respectively). In addition, the lysozyme content was high (0.15 g/100 ml) and represented about the 9% of the total proteins; this enzyme has antimicrobial activity [1]. Fat content significantly differed from human milk, thus it would require an energetic integration when is used in infant feeding. By contrast, the milk showed a fatty acid composition of nutritional interest, especially for the unsaturated fatty acid content (45 g/100 g of fatty acids). In particular, the unsaturated/saturated fatty acid ratio and the ω6/ω3 ratio were 0.82 and 1.65 respectively. Oleic, linoleic, alpha linolenic and eicosapentanoic acids were respectively 17%, 14%, 8% and 0.13% of the total fatty acids. These polyunsaturated fatty acids are known for their beneficial properties on human health. In addition, milk fat globules showed a mean diameter of 1.92 micron, smaller than the diameter reported for other milks. The smaller globule diameter may promote the digestibility of donkey milk fat [2]. The high lactose content (7.1 g/100 ml) is also of nutritional interest because of its contribution on milk palatability that makes the milk acceptable by children. Ash content was lower than cow milk and was about 0.36 g/100 ml. With regard to individual minerals, the amount of calcium (70 mg/100 ml), phosphorus (60 mg/100 ml), potassium (60 mg/100 ml) and sodium (20 mg/100 ml) were more similar to human milk. The Amiatina donkey milk had also a high content of vitamin D (2.3 µg/100 ml; 92 IU/100 ml on the average), higher in summer than in winter. In conclusion, the similarities with human milk were high, except for the fat content. This finding highlights the need for energy integrations especially when the milk is used in early childhood. Moreover, donkey milk use could be extended to obese people and to the elderly

Investigating fishmeal replacement with insect meal in "aquafeed" using zebrafish (Danio rerio) as animal model

The world population is continuously growing and it is expected to reach 9 billions of people by 2050. In this context, to secure food for everyone it is necessary to double the world food production and in the last decades, Aquaculture is already significantly increasing its production worldwide. Unfortunately, fish nutrition strictly relies on the use of fishmeal (FM) as source of protein and the intensive catch of herrings, anchovies and sardines have strongly reduced the consistence of the fish wild stocks; hence, the production of supplementary quantities of FM is considered not realistic and, in particular, not “sustainable”. As a consequence, the market price of FM have been dramatically increasing in the last decades and the feed industry and research Institutions are now strongly engaged in seeking valid alternative to this important protein source (FAO, 2014). Currently, the majority of FM replacement have been done with vegetal ingredients such as soybean, soybean extracts, corn gluten and other grains, all of them somehow characterized by an unbalanced amino acid profiles, presence of anti-nutritional factors and limited digestibility rates. In this context, thanks to its low “carbon footprint” and production sustainable, insect meal (IM) may represent a valid alternative to FM (FAO, 2013). Hence, the aim of this study was to evaluate the effects of replacing FM with IM, in fish feed. The IM used was made by Hermetia illucens larvae (Insect protein meal®, Protix©, Dongen - NL); since several Authors refer that Zebrafish (Danio rerio) can be considered a suitable animal model for aquaculture nutrition research (Dahm e Geisler, 2006; Ulloa, 2011; Ulloa 2014), it was used to carry out the present study at the zebrafish facility of the Department of Veterinary Science of the University of Pisa. In particular, 320 juveniles (Wild-Type; AB Line; body weight 85±25 mg (m±ds); 60 days old) were randomly divided into four groups and four replicates, and fed 4 diets (iso-energetic and iso-proteic) each differing for the IM/FM replacing rate, as follow: Control, no IM and 20% FM (0% FM replacement); HI25%, 5 % IM and 15% FM (25% FM replacement); HI50% 10 % IM and 10% FM 50% FM replacement); HI 100%, 20% IM, no FM (100% FM replacement). The feed, extruded, was produced by Sparos© (Olhão - P), spin-off company of the University of Algarve, and fed four time per day for a 49 days total period. The diet’s effects on fish were investigated through the growth performances, mortality rate, feed consumption and feed conversion rate; also, at the end of the experimental period, the fish intestine was sampled for performing histological analysis (villi length and diameter, globlet cells counts). All the performances and parameters observed were consistent to those normally observed on other freshwater and marine fish, this meaning that zebrafish can be a suitable animal model for studies on fish growth performances and nutrition in aquaculture. In fact, at the end of the trial, zebrafish have almost tripled their body weight. For all the considered parameters, the groups fed IM did not show differences (p&gt;0.05) within experimental groups, as well as in comparison to the control group. The final mean body weights reached by each group were: control group 229 mg; 0% 237 mg; HI50 242 mg; HI100 229 mg; feed conversion ratios were: control group 2.11; HI25 2.05; HI50 1.94; HI100 2.08. Only significant differences (p&lt;0.05) were observed between males and females fish, hence independently from the considered treatments. In conclusion, results suggest that when insect meal from Hermetia illucens is included up to 20% in Zebrafish feed, it is a valid alternative for replacing fishmeal in Zebrafish and, verisimilarly, in other fish aquacultured fish species