Use of Biotechnology to increase the content of bioactive compounds in fermented foods of plant origin

The objectives of this PhD research were: i) to evaluate the use of bread making process to increase the content of β-glucans, resistant starch, fructans, dietary fibers and phenolic compounds of kamut khorasan and wheat breads made with flours obtained from kernels at different maturation stage (at milky stage and fully ripe) and ii) to study the impact of whole grains consumption in the human gut. The fermentation and the stages of kernel development or maturation had a great impact on the amount of resistant starch, fructans and β-glucans as well as their interactions resulted highly statistically significant. The amount of fructans was high in kamut bread (2.1g/100g) at the fully ripe stage compared to wheat during industrial fermentation (baker’s yeast). The sourdough increases the content of polyphenols more than industrial fermentation especially in bread made by flour at milky stage. From the analysis of volatile compounds it resulted that the sensors of electronic nose perceived more aromatic compound in kamut products, as well as the SPME-GC-MS, thus we can assume that kamut is more aromatic than wheat, so using it in sourdough process can be a successful approach to improve the bread taste and flavor. The determination of whole grain biormakers such as alkylresorcinols and others using FIE-MS AND GC-tof-MS is a valuable alternative for further metabolic investigations. The decrease of N-acetyl-glucosamine and 3-methyl-hexanedioic acid in kamut faecal samples suggests that kamut can have a role in modulating mucus production/degradation or even gut inflammation. This work gives a new approach to the innovation strategies in bakery functional foods, that can help to choose the right or best combination between stages of kernel maturation-fermentation process and baking temperature.

By-products of the dairy farming as raw material for the biotechnology production of polyhydroxyalkanoates

I Poliidrossialcanoati (PHA) sono poliesteri completamente biodegradabili, prodotti da microrganismi come fonte di energia e di carbonio per la sintesi di nuovo materiale cellulare, utilizzando come substrato materie prime rinnovabili. Questi poliesteri sono considerati potenziali candidati per la sostituzione delle materie plastiche convenzionali. Tuttavia, i più alti costi di produzione dei PHA in confronto a quelli delle materie plastiche derivanti dal petrolio, rappresentano il principale ostacolo per la parziale sostituzione di questi ultimi con i biopolimeri. Gli alti costi sono principalmente dovuti all'utilizzo di colture microbiche pure (in cui sia presente un solo ceppo batterico) e substrati puri e costosi. Nell'ultimo decennio è stato sviluppato un processo di produzione a tre stadi alternativo e potenzialmente a minor costo, basato sull'utilizzo di colture microbiche miste (Mixed Microbials Culture, MMC) e una varietà di substrati organici a costo contenuto o nullo, quali alcuni rifiuti dell’industria agro-alimentare. Il presente studio si è concentrato sulla prima fase del processo di produzione dei PHA da colture miste, la fermentazione acidogenica, utilizzando siero di latte come fonte di carbonio per produrre acidi organici. In particolare questo lavoro ha avuto come obiettivo quello di studiare come diverse condizioni operative utilizzate nella fase di fermentazione acidogenica possono influenzare la concentrazione e il profilo degli acidi organici prodotti. Sono stati valutati anche gli effetti dei diversi profili degli acidi organici sulla fase di selezione della coltura microbica, in termini di capacità di stoccaggio di PHA e composizione polimerica.

Country-of-Origin Labeling for Processed Foods

The country-of-origin is the “nationality” of a food when it goes through customs in a foreign country, and is a “brand” when the food is for sale in a foreign market. My research on country-of-origin labeling (COOL) started from a case study on the extra virgin olive oil exported from Italy to China; the result shows that asymmetric and imperfect origin information may lead to market inefficiency, even market failure in emerging countries. Then, I used the Delphi method to conduct qualitative and systematic research on COOL; the panel of experts in food labeling and food policy was composed of 19 members in 13 countries; the most important consensus is that multiple countries of origin marking can provide accurate information about the origin of a food produced by two or more countries, avoiding misinformation for consumers. Moreover, I enhanced the research on COOL by analyzing the rules of origin and drafting a guideline for the standardization of origin marking. Finally, from the perspective of information economics I estimated the potential effect of the multiple countries of origin labeling on the business models of international trade, and analyzed the regulatory options for mandatory or voluntary COOL of main ingredients. This research provides valuable insights for the formulation of COOL policy.

Identification of CNV and QTL for productive and functional traits in dairy cattle using dense SNP chips

The thesis identify CNV structural variants as possible markers for genomic selection and identify QTL regions for Fatty Acid Content in the Italian Brown Swiss population. Additionally it maps the QTL for mastitis resistance in the Valdostana Red Pied cattle.

Effects of long-term feeding of genetically modified corn (event MON810) on the performance of lactating dairy cows

A long-term study over 25 months was conducted to evaluate the effects of genetically modified corn on performance of lactating dairy cows. Thirty-six dairy cows were assigned to two feeding groups and fed with diets based on whole-crop silage, kernels and whole-crop cobs from Bt-corn (Bt-MON810) or its isogenic not genetically modified counterpart (CON) as main components. The study included two consecutive lactations. There were no differences in the chemical composition and estimated net energy content of Bt-MON810 and CON corn components and diets. CON feed samples were negative for the presence of Cry1Ab protein, while in Bt-MON810 feed samples the Cry1Ab protein was detected. Cows fed Bt-MON810 corn had a daily Cry1Ab protein intake of 6.0 mg in the first lactation and 6.1 mg in the second lactation of the trial. Dry matter intake (DMI) was 18.8 and 20.7 kg/cow per day in the first and the second lactation of the trial, with no treatment differences. Similarly, milk yield (23.8 and 29.0 kg/cow per day in the first and the second lactation of the trial) was not affected by dietary treatment. There were no consistent effects of feeding MON810 or its isogenic CON on milk composition or body condition. Thus, the present long-term study demonstrated the compositional and nutritional equivalence of Bt-MON810 and its isogenic CON.

Socio-Spatial Relationships in Dairy Cows

Farm animals may serve as models for evaluating social networks in a controlled environment. We used an automated system to track, at fine temporal and spatial resolution (once per minute, +/- 50 cm) every individual in six herds of dairy cows (Bos taurus). We then analysed the data using social network analyses. Relationships were based on non-random attachment and avoidance relationships in respect to synchronous use and distances observed in three different functional areas (activity, feeding and lying). We found that neither synchrony nor distance between cows was strongly predictable among the three functional areas. The emerging social networks were tightly knit for attachment relationships and less dense for avoidance relationships. These networks loosened up from the feeding and lying area to the activity area, and were less dense for relationships based on synchronicity than on median distance with respect to node degree, relative size of the largest cluster, density and diameter of the network. In addition, synchronicity was higher in dyads of dairy cows that had grown up together and shared their last dry period. This last effect disappeared with increasing herd size. Dairy herds can be characterized by one strongly clustered network including most of the herd members with many non-random attachment and avoidance relationships. Closely synchronous dyads were composed of cows with more intense previous contact. The automatic tracking of a large number of individuals proved promising in acquiring the data necessary for tackling social network analyses.

The examination and identification of bite marks in foods using 3D scanning and 3D comparison methods

Bite mark analysis offers the opportunity to identify the biter based on the individual characteristics of the dentitions. Normally, the main focus is on analysing bite mark injuries on human bodies, but also, bite marks in food may play an important role in the forensic investigation of a crime. This study presents a comparison of simulated bite marks in different kinds of food with the dentitions of the presumed biter. Bite marks were produced by six adults in slices of buttered bread, apples, different kinds of Swiss chocolate and Swiss cheese. The time-lapse influence of the bite mark in food, under room temperature conditions, was also examined. For the documentation of the bite marks and the dentitions of the biters, 3D optical surface scanning technology was used. The comparison was performed using two different software packages: the ATOS modelling and analysing software and the 3D studio max animation software. The ATOS software enables an automatic computation of the deviation between the two meshes. In the present study, the bite marks and the dentitions were compared, as well as the meshes of each bite mark which were recorded in the different stages of time lapse. In the 3D studio max software, the act of biting was animated to compare the dentitions with the bite mark. The examined food recorded the individual characteristics of the dentitions very well. In all cases, the biter could be identified, and the dentitions of the other presumed biters could be excluded. The influence of the time lapse on the food depends on the kind of food and is shown on the diagrams. However, the identification of the biter could still be performed after a period of time, based on the recorded individual characteristics of the dentitions.

Comparison of the effect of a CIDR-Select synch versus a long-term CIDR based AI protocol on reproductive performance in multiparous dairy cows in Swiss dairy farms

Background Synchronization programs have become standard in the dairy industry in many countries. In Switzerland, these programs are not routinely used for groups of cows, but predominantly as a therapy for individual problem cows. The objective of this study was to compare the effect of a CIDR-Select Synch and a 12-d CIDR protocol on the pregnancy rate in healthy, multiparous dairy cows in Swiss dairy farms. Methods Cows (N = 508) were randomly assigned to CIDR-Select Synch (N = 262) or 12-d CIDR (N = 246) protocols. Cows in the CIDR-Select Synch group received a CIDR and 2.5 ml of buserelin i.m. on d 0. On d 7, the CIDR insert was removed and 5 ml of dinoprost was administered i.m.. Cows in the 12-d CIDR group received the CIDR on d 0 and it was removed on d 12 (the routine CIDR protocol in Swiss dairies). On d 0 a milk sample for progesterone analysis was taken. Cows were inseminated upon observed estrus. Pregnancy was determined at or more than 35 days after artificial insemination. As a first step, the two groups were compared as to indication for treatment, breed, stud book, stall, pasture, and farmer's business using chi square tests or Fisher's exact test. Furthermore, groups were compared as to age, DIM, number of AI's, number of cows per farm, and yearly milk yield per cow using nonparametric ANOVA. A multiple logistic model was used to relate the success of the protocols to all of the available factors; in particular treatment (CIDR-Select Synch/12-d CIDR), milk progesterone value, age, DIM, previous treatment of the uterus, previous gynecological treatment, and number of preceding inseminations. Results The pregnancy rate was higher in cows following the CIDR-Select Synch compared to the 12-d CIDR protocol (50.4% vs. 22.4%; P < 0.0001). Conclusion The CIDR-Select Synch protocol may be highly recommended for multiparous dairy cows. The reduced time span of the progesterone insert decreased the number of days open, improved the pregnancy rate compared to the 12-d CIDR protocol and the cows did not to have to be handled more often.

The NutriChip project--translating technology into nutritional knowledge

Advances in food transformation have dramatically increased the diversity of products on the market and, consequently, exposed consumers to a complex spectrum of bioactive nutrients whose potential risks and benefits have mostly not been confidently demonstrated. Therefore, tools are needed to efficiently screen products for selected physiological properties before they enter the market. NutriChip is an interdisciplinary modular project funded by the Swiss programme Nano-Tera, which groups scientists from several areas of research with the aim of developing analytical strategies that will enable functional screening of foods. The project focuses on postprandial inflammatory stress, which potentially contributes to the development of chronic inflammatory diseases. The first module of the NutriChip project is composed of three in vitro biochemical steps that mimic the digestion process, intestinal absorption, and subsequent modulation of immune cells by the bioavailable nutrients. The second module is a miniaturised form of the first module (gut-on-a-chip) that integrates a microfluidic-based cell co-culture system and super-resolution imaging technologies to provide a physiologically relevant fluid flow environment and allows sensitive real-time analysis of the products screened in vitro. The third module aims at validating the in vitro screening model by assessing the nutritional properties of selected food products in humans. Because of the immunomodulatory properties of milk as well as its amenability to technological transformation, dairy products have been selected as model foods. The NutriChip project reflects the opening of food and nutrition sciences to state-of-the-art technologies, a key step in the translation of transdisciplinary knowledge into nutritional advice.

Quantitative assessment of physiological and behavioural parameters in healthy dairy cows evoked by transcutaneous electrical nerve stimulation of the udder

The pain and distress associated with transcutaneous electrical nerve stimulation (TENS) of the udder was evaluated by treating 20 healthy dairy cows with an electrical udder stimulator. This generated a sequence of pulses (frequency: 160+/-10% impulses per second, duration 250 mus) and provided voltage ranges from 0 to 10 volts (+/-10%). Trials took place on three consecutive days, twice daily after morning and evening milking. Daily sessions were divided into two periods: (1) control (sham treatment) and (2) treatment (real treatment). Physiological (heart rate, respiratory rate, and plasma cortisol concentration) as well as ethological parameters (kicking, weight shifting, and looking backwards to udder) were defined as pain-indicating parameters and observed. Evaluation of data showed that only one parameter (kicking) was significantly increased during real treatment compared to sham treatment. It is concluded that the TENS therapy tested in this study can evoke changes in behaviour (increased kicking) consistent with an experience of pain in some cows.

[Systematic sanitation of dairy herds with mastitis caused by Staphylococcus aureus]

Several strategies are known for sanitizing dairy herd problems caused by Staphylococcus (S.) aureus. They mostly consist of general management measures but specific decision-making at an individual animal level has not been described. A sanitation program in the form of a process chart developed by the Bern Clinic for Ruminants was undertaken in 10 dairy herds with this problem. In an affected herd the cows were divided into 3 groups: healthy, suspect, infected. Three milk samples (MS), taken at two-week intervals were cultivated. The cows were grouped according to the culture results. To measure the success of the sanitation program, the key figures <> (target < 150,000 SCC/ml) and <> (limit: 150'00 SCC/ml, target < 20 %) were used. These were compared with the corresponding key figures from dairy herds, which were followed-up by the Bern Clinic for Ruminants (control herds). The problem herd sanitation program lasted between 2 and 21 months. A total of 1598 MS were analyzed, of which 241 (15 %) were S. aureus positive (15 %). At the end of the sanitation the key figures between problem herds and control herds were similar. The sanitation program has proved to be practical. The detection of S. aureus positive cows proved to be reliable and the udder health of the herd could be significantly improved.