Ester Grilli

Vaccination of Lactating Dairy Cows for the Prevention of Aflatoxin B1 Carry Over in Milk

The potential of anaflatoxin B1 (AnAFB1) conjugated to keyhole limpet hemocyanin (KLH) as a vaccine (AnAFB1-KLH) in controlling the carry over of the aflatoxin B1 (AFB1) metabolite aflatoxin M1 (AFM1) in cow milk is reported. AFB1 is the most carcinogenic compound in food and foodstuffs amongst aflatoxins (AFs). AnAFB1 is AFB1 chemically modified as AFB1-1(O-carboxymethyl) oxime. In comparison to AFB1, AnAFB1 has proven to be non-toxic in vitro to human hepatocarcinoma cells and non mutagenic to Salmonella typhimurium strains. AnAFB1-KLH was used for immunization of cows proving to induce a long lasting titer of anti-AFB1 IgG antibodies (Abs) which were cross reactive with AFB1, AFG1, and AFG2. The elicited anti-AFB1 Abs were able to hinder the secretion of AFM1 into the milk of cows continuously fed with AFB1. Vaccination of lactating animals with conjugated AnAFB1 may represent a solution to the public hazard constituted by milk and cheese contaminated with AFs.

Pediocin A improves growth performance of broilers challenged with Clostridium perfringens

The aim of this study was to investigate the efficacy of the anticlostridial pediocin A from Pediococcus pentosaceus FBB61 to contain negative effects associated to Clostridium proliferation in broilers, through 2 subsequent investigations. In the first study, 36 Ross 508 broilers were divided into 3 groups and fed for 21 d as follows: the control diet (CTR), the control diet supplemented with supernatant filtrate of a culture of P. pentosaceus FBB61-2 (Bac-, isogenic mutant nonproducing pediocin A), and the control diet supplemented with supernatant filtrate of a culture of P. pentosaceus FBB61 (Bac+) containing pediocin A. Birds were challenged with 10(6) cells of Clostridium perfringens. In the second study, 216 Ross 508 broilers were allocated in 18 pens and divided into 3 groups fed the same diet for 42 d: a control group (CTR), a group challenged with 10(8) cells of C. perfringens (CP), and a group challenged with 10(8) cells of C. perfringens and receiving the control diet supplemented with P. pentosaceus FBB61 and pediocin A (PA). Broiler BW, ADG, ADFI, and feed conversion rate were measured throughout the studies. At the end of both experiments, an appropriate number of birds was killed and analyzed for necrotic enteritis lesions and microbiological examinations. In the first study, on d 9, ADG and BW were 20% higher for Bac+ compared with CTR and Bac-; on d 14, ADG was higher for Bac+ (+23%, P<0.05), whereas BW was higher for Bac+ and Bac- compared with CTR (+23 and +14%, respectively; P<0.05). In the second study, on d 14, ADG and BW were higher for PA compared with CTR and CP (+15% on average; P<0.05), whereas between 15 and 42 d, there was only a tendency toward a higher ADG for PA when compared with the CP group (+4%, P=0.08). Diet supplementation with pediocin A improved broiler growth performance during the challenge with C. perfringens and tended to restore the ADG depletion during the 42-d period.

Microencapsulated lysine and low-protein diets: Effects on performance, carcass characteristics and nitrogen excretion in heavy growing–finishing pigs

Two studies were conducted to assess the effects of dietary protein and Lys reduction on growth performance, carcass quality, N excretion, and plasma N profile in growing-finishing pigs from 35 to 180 kg. The growing trial was conducted with 72 gilts and 72 barrows with 6 pens per treatment and 6 gilts or 6 barrows per pen. Four diets with the same DE and NE were compared: 1) control diet, 2) diet with protein content reduced by 3% units compared with the control diet and supplemented with Lys HCl to match the requirements according to the 2012 NRC (FLys), 3) diet similar to FLys for protein level but supplemented with only 50% of the Lys HCl provided with FLys (-50% FLys), and 4) diet similar to FLys and -50% Flys for protein level with microencapsulated Lys added to supply 20% of the Lys provided by FLys (-80% MLys). Pigs fed the FLys diet performed in a similar manner and showed similar carcass characteristics to the control pigs. Pigs fed the -50% FLys diet had the lightest BW (P < 0.01) as well as the lightest carcass (P = 0.02) and trimmed thigh (P = 0.04) weights. Pigs fed the -80% MLys diet showed growth performance and carcass characteristics similar to the control pigs. The N balance study was conducted with 8 barrows arranged in a double replicated 4 × 4 Latin square design. The control pigs had greater (P < 0.01) N intake and urinary and total N excretion compared with pigs fed the low-protein diets. The reduction of the dietary protein concentration by 3% units decreased total N excretion by an average of 24.5% and the greatest reduction of total N excretion was obtained by the -80% MLys diet (26.12%) compared with the control diet throughout the trial. No effect of dietary treatment was observed on the plasma concentrations of ammonia, urea, and total AA in the 80-kg pigs. Decreased plasma ammonia (P < 0.01) and urea (P = 0.03) concentrations were detected in the 120-kg pigs fed the low-protein diets. The 160-kg pigs fed the -80% MLys pigs had less total AA, indispensable AA in particular (P < 0.01), than pigs fed the other diets and decreased ammonia (P < 0.01) and urea (P = 0.05) concentrations than the control pigs. The results showed that the use of microencapsulated Lys, compared with both Lys HCl and dietary protein-bound Lys, can save CP and synthetic AA in diet formulation and can reduce N excretion in manure without adversely affecting the growth performance and carcass quality of heavy growing-finishing pigs.

Influence of maturity on alfalfa hay nutritional fractions and indigestible fiber content

This study focused on changes in fibrous and protein fractions, changes in fiber digestibility and amount of indigestible neutral detergent fiber (NDF) as a consequence of increased maturity in alfalfa. A total area of 720 m(2) was divided in 18 blocks randomly assigned to 3 treatments, differing in cutting intervals. Treatment 1 was harvested with a 21-d cutting schedule, at a prebloom stage; treatment 2 with a 28-d schedule, at about first-bloom stage; whereas a full bloom was observed in treatment 3, harvested with a 35-d cutting schedule. Treatments were replicated 4 times through the spring-summer period for 2 subsequent years, 2011 and 2012. Statistical differences were observed for crude protein [treatment 1: 20.8%, treatment 2: 17.3%, and treatment 3: 17.0%; standard error of the mean (SEM)=0.83], soluble protein, and nonprotein nitrogen among treatments on a dry matter basis. Similar results were observed for acid detergent lignin (6.3, 6.9, and 7.3%, respectively; SEM=0.39), lower in treatment 1 compared with others, and in vitro NDF digestibility at 24 or 240 h. Indigestible NDF at 240 h resulted in lower values for treatment 1 compared with treatments 2 and 3 (15.5, 17.2, and 18.3%, respectively; SEM=1.54). Moreover, the indigestible NDF:acid detergent lignin ratio varied numerically but not statistically among treatments, being as much as 9% greater than the 2.4 fixed value applied for rate of digestion calculation and Cornell Net Carbohydrate Protein System (Cornell University, Ithaca, NY)-based model equations. Assuming the diet composition remained unchanged, treatment 3 (35-d cutting interval) would be expected to yield 1.4 kg less milk per day based on energy supply, and 2.8 kg less milk daily based on protein supply than treatment 1.

Pediocin A modulates intestinal microflora metabolism in swine in vitro intestinal fermentations.

The aim of this study was to investigate in vitro the effects of pediocin A [a bacteriocin produced by lactic acid bacteria (LAB) Pediococcus pentosaceus FBB61] on microbial metabolism in the small and large intestine of pigs. Pediocin A was partially purified by ion exchange chromatography and added to an in vitro fermentation system. The intestinal inoculum was collected from pigs immediately after slaughter, diluted with a buffer, and dispensed into fermentation syringes and vessels of the 2 experimental groups: 1) Bac+ = cecal liquor + predigested diet + pediocin A (final concentration 160 activity units/mL); 2) Bac- = cecal liquor + predigested diet + partially purified supernatant of P. pentosaceus FBB61-2. Intestinal microbial growth was monitored using the cumulative gas production technique; the kinetics of fermentation, bacterial counts, VFA, ammonia, polyamines, and p-cresol production were analyzed. Pediocin A had almost no effects on small intestine fermentation parameters, whereas in the cecum pediocin A decreased gas production (-16%; P < 0.05), ammonia, and VFA production (-52 and -21%, respectively, after 24 h; P < 0.001) compared with the control group. Significant inhibition of clostridia and LAB occurred in cecal fermentations: the Bac+ group yielded a decreased number of clostridia and LAB in cecal fermentations (8.19 and 7.80 cfu/mL, respectively) compared with Bac- (9.32 and 8.95 cfu/mL, respectively; P < 0.001). The low clostridia counts in the pediocin-treated group may also explain the reduced concentration of the carcinogenic compound p-cresol (-88%; P < 0.01). Our results suggest that pediocin A could be an alternative to replace antibiotic growth promoters for the prophylaxis of enteric diseases and to improve production of farm animals.

Intestinal metabolism of weaned piglets fed a typical United States or European diet with or without supplementation of tributyrin and lactitol

The aim of the study was to investigate the effects of supplementation of a microencapsulated blend of tributyrin and lactitol (TL) to a standard European (EU) diet without antibiotic growth promoters on intestinal metabolism and mucosa development of weaned piglets and to compare it with a standard US diet containing animal proteins, zinc oxide, copper sulfate, and carbadox. Ninety piglets weaned at 21 d were divided into 3 dietary groups consisting of 5 replicates each: 1) US diet supplemented with 55 mg/kg of carbadox, and 2.5% each of plasma proteins and spray-dried blood cells in the first phase, 3,055 mg/kg of Zn in the first and second phases, and 180 mg/kg of Cu in the third phase; 2) EU diet based on vegetable proteins and no antibiotics; and 3) the same EU diet supplemented with 3,000 mg/kg of microencapsulated TL. The study was divided into 3 phases: 0 to 7, 8 to 21, and 22 to 35 d. On d 7, 21, and 35, animals were weighed, and feed consumption and efficiency were determined. On d 14 and 35, one pig per pen was killed, and the intestinal contents and mucosa from the proximal, middle, distal jejunum and the ileum were sampled. Intestinal wall sections were fixed for histological analysis, and intestinal content was used for VFA, ammonia, and polyamine analysis. Throughout the study (d 0 to 35), the US diet had greater ADG and ADFI than the EU diet (P < 0.05). The EU diet supplemented with TL tended to have 11% greater ADG (P = 0.17). Feeding the EU diet caused a reduction in proximal and middle jejunum villi length by 10% (P < 0.05) and an increase in crypt size in proximal jejunum (P < 0.05) compared with the US diet, probably due to an increased rate of cell loss and crypt cell production. The TL supplementation resulted in longer villi along the jejunum and less deep crypts in the proximal jejunum (+15.9 and -8.9%, respectively; P < 0.05) than the unsupplemented EU diet. The TL diet increased the concentrations of cadaverine and putrescine in the small intestine (P < 0.05) and seemed to increase cadaverine, histamine, putrescine, and spermine in the large intestine by 1.5- to 10-fold compared with the US or EU diet. In conclusion, although the US diet had a greater effect on growth performance and mucosal trophic status than the EU diets, the supplementation with slowly released TL seemed to be an effective tool to partially overcome the adverse effects of vegetable protein diets.

Arcobacter butzleri, Arcobacter cryaerophilus, and Arcobacter skirrowii Circulation in a Dairy Farm and Sources of Milk Contamination.

ABSTRACT Even though dairy cows are known carriers of Arcobacter species and raw or minimally processed foods are recognized as the main sources of human Arcobacter infections in industrialized countries, data on Arcobacter excretion patterns in cows and in milk are scant. This study aimed to identify potentially pathogenic Arcobacter species in a dairy herd and to investigate the routes of Arcobacter transmission among animals and the potential sources of cattle infection and milk contamination. A strategy of sampling the same 50 dairy animals, feed, water, and milk every month for a 10-month period, as well as the sampling of quarter milk, animal teats, the milking environment, and animals living on the farm (pigeons and cats), was used to evaluate, by pulsed-field gel electrophoresis (PFGE), the characteristic patterns in animals, their living environment, and the raw milk they produced. Of the 463 samples collected, 105 (22.6%) were positive for Arcobacter spp. by culture examination. All the matrices except quarter milk and pigeon gut samples were positive, with prevalences ranging from 15 to 83% depending on the sample. Only three Arcobacter species, Arcobacter cryaerophilus (54.2%), A. butzleri (34.2%), and A. skirrowii (32.3%), were detected. PFGE analysis of 370 isolates from positive samples provided strong evidence of Arcobacter circulation in the herd: cattle likely acquire the microorganisms by orofecal transmission, either by direct contact or from the environment, or both. Water appears to be a major source of animal infection. Raw milk produced by the farm and collected from a bulk tank was frequently contaminated (80%) by A. butzleri; our PFGE findings excluded primary contamination of milk, whereas teats and milking machine surfaces could be sources of Arcobacter milk contamination.

Dimethylarginine Dimethylaminohydrolase/Nitric Oxide Synthase Pathway in Liver and Kidney: Protective Effect of Cyanidin 3-O-β-D-Glucoside on Ochratoxin-A Toxicity

The aim of the present study was to evaluate the effect of long-term cyanidin 3-O-β-D-glucoside (C3G) and/or Ochratoxin A (OTA)-exposure on dimethylarginine dimethylamino hydrolase/nitric oxide synthase (DDAH/NOS) pathway in rats. The experiments were performed in rats supplemented with C3G (1 g/kg feed), OTA (200 ppb), and OTA + C3G. After 4 weeks of daily treatment, liver and kidneys were processed for eNOS, iNOS and DDAH-1 Western blotting, nitrite levels evaluation and DDAH activity determination. Results show that OTA is able to induce iNOS both in kidney and liver, whereas OTA is able to induce eNOS and DDAH-1 overexpression and DDAH activation only in kidney, resulting in increased nitrite levels. In kidney of OTA + C3G fed rats, iNOS, eNOS and DDAH-1 expression were less pronounced compared with those observed in the OTA-treated group. Coherent with the decreased iNOS, eNOS and DDAH-1 expression a decrease in nitrite levels and DDAH activity was observed in the OTA + C3G group. Results demonstrate that C3G is able to counteract the deleterious effects of chronic consumption of OTA and also suggest a possible involvement of iNOS-eNOS-DDAH impairment in OTA nephrocarcinogenity.

Microencapsulated sorbic acid and nature-identical compounds reduced Salmonella Hadar and Salmonella Enteritidis colonization in experimentally infected chickens

The reduction of Salmonella prevalence in broilers is a priority in European Union agricultural policies because treatment with antibiotics is forbidden by Regulation (EC) 2160/2003. Two trials were conducted to evaluate the efficacy of a microencapsulated blend of sorbic acid and nature-identical compounds (i.e., chemically synthesized botanicals; SAB) on the reduction of the cecal prevalence and contents of Salmonella enterica serovars Hadar and Enteritidis in experimentally infected chickens. In the first trial, 125 one-day-old Lohmann specific-pathogen-free chickens were assigned to one of the following treatments: negative control (not challenged and not treated), positive control (challenged and not treated), SAB0.3, SAB1, or SAB5 (challenged and treated with the microencapsulated blend included in the feed at 0.03, 0.1, or 0.5%, respectively). At 30 d of age, birds were infected with 10(6) cfu of Salmonella Hadar, and after 5, 10, or 20 d postinfection, 5, 10, and 10 birds per treatment, respectively, were killed and the cecal contents and liver and spleen samples were analyzed for Salmonella Hadar. In the second trial, 100 one-day-old Ross 708 chickens were assigned to 1 of 5 treatments: control (not treated), SAB0.3, SAB1, SAB2, or SAB5 (treated with the blend included in the feed at 0.03, 0.1, 0.2, or 0.5%, respectively). At 7 d of age, the birds were challenged with 10(5) cfu of Salmonella Enteritidis, and after 7, 14, or 24 d after challenge, 5, 5, and 10 birds per treatment, respectively, were killed and cecal contents were analyzed for Salmonella Enteritidis. Results showed that in the early stage of infection Salmonella prevalence was high in both studies, whereas at the end of the observation periods, the blends at 0.03, 0.1, and 0.5 in the challenge with Salmonella Hadar and at 0.2 and 0.5% in the challenge with Salmonella Enteritidis significantly reduced (by 2 log(10) cfu) the cecal content of Salmonella. This study showed that intestinal delivery of microencapsulated sorbic acid and nature-identical compounds can result in a 100-fold reduction of Salmonella at the intestinal level in broilers at slaughter age.

Role of Benzoic, Lactic and Sorbic Acid in In Vitro Swine Cecal Fermentation

Piva, A. and Grilli, E., 2007. Role of benzoic, lactic and sorbic acid in in vitro swine cecal fermentation. Veterinary Research Communications, 31(Suppl. 1), 401–404