Effects of Supplementation of a Mycotoxin Mitigation Feed Additive in Lactating Dairy Cows Fed Fusarium Mycotoxin-Contaminated Diet for an Extended Period.

Fusarium mycotoxins are inactivated by rumen flora; however, a certain amount can pass the rumen and reticulum or be converted into biological active metabolites. Limited scientific evidence is available on the impact and mitigation of Fusarium mycotoxins on dairy cows' performance and health, particularly when cows are exposed for an extended period (more than 2 months). The available information related to these mycotoxin effects on milk cheese-making parameters is also very poor. The objective of this study was to evaluate a commercially available mycotoxin mitigation product (MMP, i.e., TOXO® HP-R, Selko, Tilburg, The Netherlands) in lactating dairy cows fed a Fusarium mycotoxin-contaminated diet, and the repercussions on the dry matter intake, milk yield, milk quality, cheese-making traits and health status of cows. The MMP contains smectite clays, yeast cell walls and antioxidants. In the study, 36 lactating Holstein cows were grouped based on the number of days of producing milk, milk yield, body condition score and those randomly assigned to specific treatments. The study ran over 2 periods (March/May-May/July 2022). In each period, six animals/treatment were considered. The experimental periods consisted of 9 days of adaptation and 54 days of exposure. The physical activity, rumination time, daily milk production and milk quality were measured. The cows were fed once daily with the same total mixed ration (TMR) composition. The experimental groups consisted of a control (CTR) diet, with a TMR with low contamination, high moisture corn (HMC), and beet pulp; a mycotoxins (MTX) diet, with a TMR with highly contaminated HMC, and beet pulp; and an MTX diet supplemented with 100 g/cow/day of the mycotoxin mitigation product (MMP). The trial has shown that the use of MMP reduced the mycotoxin's negative effects on the milk yield and quality (protein, casein and lactose). The MTX diet had a lower milk yield and feed efficiency than the CTR and MMP HP-R diets. The MMP limited the negative effect of mycotoxin contamination on clotting parameters, mitigating the variations on some coagulation properties; however, the MMP inclusion tended to decrease the protein and apparent starch digestibility of the diet. These results provide a better understanding of mycotoxin risk on dairy cows' performances and milk quality. The inclusion of an MMP product mitigated some negative effects of the Fusarium mycotoxin contamination during this trial. The major effects were on the milk yield and quality in both the experimental periods. These results provide better insight on the effects of mycotoxins on the performance and quality of milk, as well as the cheese-making traits. Further analyses should be carried out to evaluate MMP's outcome on immune-metabolic responses and diet digestibility.

Blood calcium dynamics in cows receiving an aqueous calcium suspension for voluntary consumption or a calcium bolus following parturition.

The form of oral calcium (Ca) supplement and the Ca source influence Ca absorption dynamics resulting in different postpartum calcemia. The objective of this study was to investigate whether an oral Ca supplement (mainly CaCO3) offered for voluntary consumption would maintain or increase postpartum blood Ca to the same degree as a Ca bolus (mainly CaCl2) providing an equivalent dose of a Ca. A total of 72 Holstein cows were blocked by expected parturition date and parity. Within each block of 3 animals, cows were randomly assigned to one of three treatments, including an oral Ca supplement offered for voluntary consumption (Ca-drink, n = 23), an oral Ca bolus (Ca-bolus, n = 24), or an untreated group (CON, n = 25). Treatments were administered once within 15 min postpartum. The Ca-drink provided 45 g of Ca (CaCO3 source) and was mixed in 20 L of lukewarm water and offered to cows for 30 min. The Ca-bolus provided 43 g of Ca (71% from CaCl2 and 29% from CaSO4) and was administered once. Both Ca-bolus and CON cows received 20-l of lukewarm water at parturition to standardize the volume of fluids (Ca-drink or 20-l lukewarm water) offered at parturition. Dairy cows offered Ca-drink had a 28% higher fluid consumption than Ca-bolus and CON cows. Milk yield and milk composition expressed in percentage protein, fat, lactose, and urea did not differ, whilst there was a small but significant increase in DMI in cows receiving the Ca-drink compared to CON, while Ca-bolus did not differ from other groups. This was consistent with reduced BW losses between week 1 and 3 in cows receiving the Ca-drink suspension. Treatment by time interactions were present for blood Ca, glucose, and urea concentrations. Blood Ca was relatively stable in Ca-drink cows, while higher fluctuations were observed in Ca-bolus cows. In Ca-bolus cows, blood Ca increased from 15 min to 6 h, decreased from 6 to 24 h, and finally increased again from 24 to 48 h. At 24 h post administration, blood Ca was greater in cows receiving the Ca-drink than cows receiving the Ca-bolus. Blood glucose was greater in Ca-bolus cows at 15 min after treatment administration compared with Ca-bolus and CON, while blood urea was higher in CON than Ca-drink and Ca-bolus throughout the sampling period. These results indicate that voluntary oral Ca resulted in a relatively stable calcemia, whereas higher fluctuations were observed in cows receiving the Ca-bolus. Due to a lack of differences between Ca-drink and Ca-bolus compared with CON, it is not possible to conclude regarding the efficacy in maintaining postpartum blood Ca.

Strategies of inorganic and organic trace mineral supplementation in gestating hyperprolific sow diets: effects on the offspring performance and fetal programming.

The aim of the present study was to evaluate the effect of trace mineral nutrition on sow performance, mineral content, and intestinal gene expression of neonate piglets when inorganic mineral sources (ITM) were partially replaced by their organic mineral (OTM) counterparts. At 35 d postmating, under commercial conditions, a total of 240 hyperprolific multiparous sows were allocated into three experimental diets: 1) ITM: with Zn, Cu, and Mn at 80, 15, and 60 mg/kg, respectively; 2) partial replacement trace mineral source (Replace): with a 30 % replacement of ITM by OTM, resulting in ITM + OTM supplementation of Zn (56 + 24 mg/kg), Cu (10.5 + 4.5 mg/kg), and Mn (42 + 18 mg/kg); and 3) Reduce and replace mineral source (R&R): reducing a 50% of the ITM source of Zn (40 + 24 mg/kg), Cu (7.5 + 4.5 mg/kg), and Mn (30 + 18 mg/kg). At farrowing, 40 piglets were selected, based on birth weight (light: <800 g, and average: >1,200 g), for sampling. Since the present study aimed to reflect results under commercial conditions, it was difficult to get an equal parity number between the experimental diets. Overall, no differences between experimental diets on sow reproductive performance were observed. Light piglets had a lower mineral content (P < 0.05) and a downregulation of several genes (P < 0.10) involved in physiological functions compared with their average littermates. Neonate piglets born from Replace sows had an upregulation of genes involved in functions like immunity and gut barrier, compared with those born from ITM sows (P < 0.10), particularly in light piglets. In conclusion, the partial replacement of ITM by their OTM counterparts represents an alternative to the totally inorganic supplementation with improvements on neonate piglet gene expression, particularly in the smallest piglets of the litter. The lower trace mineral storage together with the greater downregulation of gut health genes exposed the immaturity and vulnerability of small piglets.

Effects of two zinc supplementation levels and two zinc and copper sources with different solubility characteristics on the growth performance, carcass characteristics and digestibility of growing-finishing pigs.

The present study was conducted to evaluate the effect of two Zn supplemented levels and two Zn and Cu sources (sulphate and hydroxychloride) on growing-finishing pigs. An in vitro study and an in vivo study were conducted. In the in vitro study, Zn solubility from each source at different Zn supplementation levels was evaluated, as well as the phytic phosphorus (PP) solubility derived from the interaction or not with phytic acid at similar conditions to those found in digestive tract. The most critical interaction of Zn with phytic acid was at pH 6.5 and with Zn sulphate, resulting in the reduction in PP solubility. In the in vivo experiment, a total of 444 pigs ([Duroc × Landrace]×Pietrain; initial BW: 18.7 ± 0.20 kg) were allotted to 36 pens in a randomized complete block design (2 × 2) factorial arrangement with two Zn and Cu sources and two Zn supplemental levels (20 and 80 mg/kg). The Cu supplementation was fixed at 15 mg/kg for all diets. There was no effect of the interaction between mineral source × Zn level or Zn level on growth performance or carcass characteristics (p > .10). Apparent total digestibility of Zn and Cu along with carcass yield was higher for pigs fed hydroxychloride than pigs fed the sulphate counterparts (p < .05). Feeding low levels of Zn decreased Zn (45.5%; p < .0001) and Cu(18.5%; p = .018) faecal excretion. In conclusion, under commercial conditions, feeding growing-finishing pigs with Zn levels below those established by the European Union regulation did not affect growth performance and carcass characteristics. Reducing dietary mineral (Zn and Cu) diet content resulted in a lower faecal mineral excretion. Pigs fed sulphate minerals had an improved performance during grower period, while pigs fed hydroxychloride minerals showed an improved performance during finishing period and a greater carcass yield and mineral digestibility than those fed sulphates.

Dietary Preference of Newly Weaned Pigs and Nutrient Interactions According to Copper Levels and Sources with Different Solubility Characteristics.

Two feeding preference experiments and an in vitro assay were performed to assess the weaned pig preference for Cu doses and sources based on their sensorial perception and on the likely post-ingestive effects of Cu. At day 7 post-weaning, a total of 828 pigs were distributed into two different experiments. In Exp.1 (dose preference) a diet with a nutritional Cu level (15 mg/kg) of Cu sulfate (SF) was pair offered with higher Cu levels (150 mg/kg) of either SF or hydroxychloride (HCl). In Exp.2 (source preference), a diet supplemented with Cu-SF at 150 mg/kg was compared to a Cu-HCl (150 mg/kg) diet. At the short-term (day 7-9) and for the entire experimental week (day 7-14), pigs preferred diets with a high Cu level than with Cu at a nutritional dose (p < 0.05). Likewise, pigs preferred diets supplemented with a Cu-HCl source compared to diets with Cu-SF (p < 0.05). In vitro assay results showed a greater solubility and interaction of Cu-SF with phytic acid compared to Cu-HCl. In conclusion, pigs chose diets with higher levels of Cu probably to re-establish homeostasis after weaning. Pigs preferred diets with Cu-HCl compared to Cu-SF probably due to their solubilities and chemical differences.

Sulfate and hydroxychloride trace minerals in poultry diets - comparative effects on egg production and quality in laying hens, and growth performance and oxidative stress response in broilers.

Two experiments investigated the effect of sulfate and hydroxychloride trace minerals (TM), Zn, Cu, and Mn, in laying hens and broiler chickens. In Expt. 1, Lohmann Brown pullets (total of 1,344) at 21 wk of age were used for a 24-wk experiment. Each of the two treatments had 32 replicates with 21 hens per replicate. At 45 wk of age, three eggs per cage were randomly selected and used for internal quality assessment. In Expt. 2, Ross 308 broilers (total of 1,080) were allocated to two treatments. Each treatment had 30 replicates with 15 chicks per replicate. On day 28, after weighing, three birds were randomly selected from 15 randomly selected pens per treatment. The birds were euthanized and blood was collected for analysis for uric acid, C-reactive protein and methylmalonic acid. Samples were also taken from pectoralis muscle of each chicken and analyzed for mRNA expression of protein synthesis or hydrolysis genes. On day 35, 7 birds per pen were used for carcass evaluation. In Expt. 1, egg weight was greater (P < 0.01) in birds receiving sulfate TM from week 16 (of experiment) onwards whereas the percentage of cracked eggs was lower (P < 0.01) in hens receiving hydroxychloride TM. Percentage hen-day production tended to be greater (P < 0.10) in hens receiving hydroxychloride TM in weeks 4 to 8 only. In Expt. 2, birds receiving hydroxychloride TM had greater (P < 0.05) weight gain and tended to have greater (P < 0.10) feed intake on day 35. Expression of the gene, PSMA1, was lower (P < 0.05) whereas plasma level of uric acid and methyl malonic acid tended to be lower (P < 0.10) in birds receiving hydroxychloride TM. It was concluded that hydroxychloride TM reduced egg loss in hens at peak production and that improved growth performance response in broilers can be partly explained by reduction in proteolytic activities in the pectoralis muscle and greater resilience to oxidative stress.

Copper and zinc sources and levels of zinc inclusion influence growth performance, tissue trace mineral content, and carcass yield of broiler chickens.

A 35-d experiment was conducted in broilers to study the effect of supplementation of sulfate or hydroxychloride forms of Zn and Cu at 2 supplemental Zn levels on growth performance, meat yield, and tissue levels of Zn. On day 0, 900 male Ross 308 broiler chicks (45 ± 1.10 g) were allocated to 4 treatments in a randomized complete block design and 2 × 2 factorial arrangement of treatments. The factors were 2 sources (sulfate or hydroxychloride) of Zn and Cu and 2 levels (low or high) of Zn. The Zn sources were zinc sulfate monohydrate (ZSM) or hydroxychloride Zn. Copper sources were copper (II) sulfate pentahydrate or hydroxychloride Cu. Each of the 4 treatments had 15 replicates and 15 birds per replicate. Birds were weighed on days 0, 21, and 35 for growth performance. On day 35, left tibia bone, liver, and blood were collected from 4 randomly selected birds per pen. In addition, 7 birds per pen were used for carcass evaluation. There was no significant source × level interaction on any of the growth performance response. Broiler chickens receiving hydroxychloride Zn and Cu had greater (P < 0.05) gain: feed, whereas broiler chickens receiving lower Zn level had greater (P < 0.01) weight gain. There was no source × level interaction on meat yield. Broiler chickens receiving hydroxychloride Zn and Cu had greater (P < 0.05) % breast yield than those receiving sulfate Zn and Cu. Higher level of Zn, irrespective of source, produced greater (P < 0.01) tibia and plasma Zn levels, whereas liver Cu was greater (P < 0.05) in broiler chickens receiving hydroxychloride Zn and Cu. It was concluded that hydroxychloride Zn and Cu were more efficacious than sulfate Zn and Cu in promoting growth performance and enhancing meat yield in the current study.

A combined approach of vesicle formulations and microneedle arrays for transcutaneous immunization against hepatitis B virus.

In the search for an optimal approach for the transcutaneous immunization (TCI) of hepatitis B surface antigen (HBsAg), two vesicle formulations, L595 vesicles (composed of sucrose-laurate ester and octaoxyethylene-laurate ester) and sPC vesicles (composed of soybean-phosphatidylcholine and Span-80) were prepared and characterized in vitro and in vivo. HBsAg was associated to the vesicles, resulting in sPC-HBsAg vesicles (±170nm) with 79% HBsAg association and L595-HBsAg vesicles (±75nm) with only 29% HBsAg association. The vesicles induced in mice via TCI an antibody response only when the skin was pretreated with microneedles. This response was improved by the adjuvant cholera toxin. The sPC-HBsAg vesicle formulations showed to be the most immunogenic for TCI, which was related to the higher degree of HBsAg association.

Bioassay based screening of steroid derivatives in animal feed and supplements.

Receptor binding transcription activation bioassays are valuable tools for the screening of steroid hormones in animal feed and supplements. However, steroid derivatives often lack affinity for their cognate receptor and do not show any direct hormonal activity by themselves. These compounds are thus not detected by these kinds of bioassays and need a bioactivation step in order to become active, both in vivo and in vitro. In this study a comparison was made between different in vitro activation methods for hormone esters and hormone glycosides. Testosterone acetate and testosterone decanoate were chosen as model compounds for the hormone esters, representing the broad range of steroid esters of varying polarities, while genistin was used as a substitute model for the steroid-glycosides. Concerning bioactivation of the steroids esters, the efficiency for alkaline hydrolysis was 90-100% and much better as compared to enzymatic deconjugation by esterase. As a result 1 µg testosterone ester per gram of animal feed could easily be detected by a yeast androgen bioassay. When comparing different enzyme fractions for deglycosilation, genistin was shown to be deconjugated most efficiently by ß-glucuronidase/aryl sulfatase from Helix pomatia, resulting in a significant increase of estrogenic activity as determined by a yeast estrogen bioassay. In conclusion, chemical and enzymatic deconjugation procedures for ester and glycoside conjugates respectively, resulted in a significant increase in hormonal activity as shown by the bioassay readouts and allowed effective screening of these derivatives in animal feed and feed supplements.