Search

Highlights • Low serum 25-hydroxyvitamin D levels are detected in Japanese Akita dogs. • Healthy Akitas show a higher level of vitamin D than pathological ones. • Depigmented skin areas appearance is associated with low levels of vitamin D. • Sebaceous adenitis is the prevalent disease recorded among investigated Akitas., Low serum level of vitamin D has been linked to numerous diseases. The Akita breed of dog shows a high incidence of immune-mediated diseases such as uveodermatologic (UVD) syndrome and sebaceous adenitis (SA). Furthermore, in Japanese Akita dogs, skin depigmentation often appears after birth at different ages, in healthy dogs, and without any other symptoms. In humans poliosis is associated to low vitamin D level. The aim of this survey was to evaluate serum 25-hydroxyvitamin D (25(OH)D) levels in Japanese Akita dogs. One hundred and three Akita blood samples were collected for the 25(OH)D evaluation. Age, sex, current disease, presence and location of skin depigmentation, and diet were reported on a medical record. Based on history and clinical examination, dogs were classified according their health status as healthy (HE) or pathological (PA). The overall mean of 25(OH)D concentration was 82.42 ± 26.14 ng/mL, whereas the concentrations in the healthy dogs were significantly higher (84.90 ± 3.36 ng/mL) in comparison with the dogs with diseases (72.13 ± 4.09 ng/mL) (P

Lactobacillus rhamnosus GG is a probiotic for humans and is normally not found in pigs; however, it has been shown to protect the human-derived intestinal Caco-2 cells against the damage induced by an important intestinal pathogen, enterotoxigenic Escherichia coli F4 (ETEC). An experiment was conducted to test whether the dietary addition of Lactobacillus rhamnosus GG improves the growth and health of weaned pigs when orally challenged by E. coli F4. Thirty-six pigs were weaned at 21 days and assigned to a standard weaning diet with or without 1010 CFU Lactobacillus rhamnosus GG (ATCC 53103) (LGG) per day. The pigs, individually penned, were orally challenged with 1.5 ml of a 1010 CFU E. coli F4 suspension on day 7 and slaughtered on day 12 or day 14. With the addition of LGG, the average daily gain and the average daily feed intake were reduced after the challenge with ETEC and for the entire trial (P

The study addresses the first two weeks of piglet weaning and is aimed at i) identifying new probiotic strains of Bifidobacterium spp. and non-digestible oligosaccharides (NDO) able to increase the level of autochthonous bifidobacteria in the gut, and ii) finding the best probiotic doses, and probiotic-NDO combinations for piglet growth and gut health. Initially, on a total of 60 pigs we administered 12 strains of 4 bifidobacteria species taken from our collection of strains of different ecological origin. We found the most promising probiotic candidates to be two strains of B. animalis subsp. lactis (M 354 and Ra 18) and one strain of B. choerinum (Su 891), as these led to an increase in the number of viable bifidobacteria in the caecum. In a second trial, a different set of 64 piglets were fed a diet containing a galacto-oligosaccharide (1%) from milk whey, and two fructo-oligosaccharides (4%) from chicory inulin and sugar beet (SbFOS). There was virtually no effect on the hindgut microbiota, except for SbFOS at 4% which tended to increase the endogenous bifidobacteria. Then, in a third trial employing 128 weaning pigs, the probiotic strains Ra 18 and Su 891 were tested individually, at different doses (0; 10 7 ; 10 9 ; 10 11 per pig per day) and combined with 0% or 2% SbFOS supplementation. It was found that Ra 18 improved the live weight growth linearly, while the Su 891 did not. The FOS supplementation showed no interaction with the Ra 18 or Su 891 probiotics for growth performance and no effect on this response criterion. Our selection procedure has highlighted that B. animalis subsp. lactis Ra 18, at 10 11 CFU per pig per day, appears to be the best probiotic choice for improved growth performance in weaning piglets.

Our aim was to evaluate the ability of different strains of Bifidobacteria to cross the gut wall and reach the liver of weaning pigs, and to study the influence of different doses of a probiotic on this parameter. The ability of bacteria to pass from the intestinal lumen to mesenteric lymph nodes and other tissues is called translocation. Several factors promote bacterial translocation, including intestinal bacterial overgrowth, deficiencies in host immune response, and gut barrier damage. Translocation can also be used as a parameter to evaluate a candidate probiotic. Few data are available on the probiotic translocation in weaning pig. We analysed the liver samples from two trials performed with 32 and 64 pigs, respectively, reared from 21 to 35 days of age, fed seven different diets: control diet (C), or C supplemented with one of six different Bifidobacteria strains (1010/CFU/d) (trial 1); or supplemented with four different levels of B. animalis (0, 107, 109, 1011) crossed with 0% or 2% sugar beet fructo-oligosaccharides (trial 2). We found Bifidobacterium spp. genus-specific DNA in liver of subjects from all dietary treatments (control fed pigs included), and no relationship between oral dose and strain appeared. On average 53.1% (trial 1) and 48.4% (trial 2) of the pigs showed the presence of Bifidobacterium spp. genus-specific DNA. We conclude that both endogen and exogenous bacteria can translocate to the liver.

The aim of this study was to determine the effects of 3 antibiotics used for pulmonary pathologies added in the feed of weaned pigs on growth performance, commensal microbiota, and immune response. At weaning, a total of 72 pigs were randomly assigned by BW and litter to 1 of the following diets: control (typical weaning diet), control + 400 mg of tilmicosin/kg, control + 600 mg of amoxicillin/kg, and control + 300 mg of doxycycline/kg. Individually penned pigs were slaughtered after 3 wk (12 pigs/treatment) or 4 wk (6 pigs/treatment). During the fourth week, all pigs received the control diet to test the residual effect of the antimicrobial supplementation. The antibiotic supplementation increased growth and feed intake during the first week (P < 0.01) and over the first 3 wk combined (P < 0.05). Gain-to-feed ratio tended to improve during the first week (P = 0.076) by the antibiotics compared with the control. Among the antibiotic treatments, no difference was observed in ADG and feed intake, which were also unchanged by the diet in the fourth week. The fecal enterobacteria counts were increased by amoxicillin on d 14 and 21 (P < 0.05 and 0.01, respectively) and were decreased by tilmicosin (P < 0.001) compared with the control. Amoxicillin decreased lactic acid bacteria (P < 0.01) counts compared with the control. The antibiotic supplementation tended to decrease total bacteria variability in the jejunum (Shannon index, P = 0.091) compared with the control. The antibiotic treatment decreased the mean total serum IgM concentration (P = 0.016) after 3 wk and did not change the mucosal histomorphometry of the small intestine. For tilmicosin, the observed positive action on piglet performance and feed intake can originate by the decreased costs of immune activation determined by the action on intestinal microbiota. For amoxicillin and doxycycline, the observation on intestinal and fecal microbiota seems to be not sufficient to explain their growth-promoting effect.

Ingested nitrate is absorbed in the small intestine, recirculated into the saliva and reduced to nitrite by oral bacteria. In pigs receiving a moderate dietary addition of nitrate, the recirculation into the saliva is modest, so we aimed to assess the effect of higher nitrate doses to find out how the animal reacts to this new situation and to evaluate if a higher nitrate level could enhance the nitrate reduction process, improving the nitrite production Trial 1. Six piglets received 100 g of a commercial diet with 2.45% KNO(3) . In relation to baseline values, nitrate in blood serum and saliva increased 15 times, and declined after 6 h vs. 2 h. Salivary nitrite increased seven times after the addition and declined after 6 h vs. 2 h. Trial 2. Six piglets were fed a diet with or without 1.22% KNO(3) for 2 weeks. Salivary nitrate and nitrite increased with the addition of KNO3: nitrate increased from d0 to the end of the trial, nitrite increased 15 times after 1 week, but decreased after 2 weeks to 4.5-fold the control. After 2 weeks, nitrate reduced Shan diversity index of salivary microbiota. The present results indicate that the long exposure to high quantities of nitrates impairs the oral reduction of nitrate to nitrite and engenders a reduction of the mouth's microbiota diversity.

We tested the effect of Trp addition to a standard weaning diet and oral challenge with enterotoxigenic Escherichia coli K88 (ETEC) on growth and health of piglets susceptible or nonsusceptible to the intestinal adhesion of ETEC. Sixty-four pigs weaned at 21 d of age were divided into 3 groups based on their ancestry and BW: a control group of 8 pigs fed a basal diet (B), the first challenged group of 28 pigs fed B diet (BCh), and the second challenged group of 28 pigs fed a diet with Trp (TrpCh). The Trp diet was produced by the addition of 1 g of l-Trp/kg to the basal diet. On d 5, pigs were orally challenged with 1.5 mL suspension containing 10(10) cfu ETEC/mL or placebo, and killed on d 9 or 23. Based on in vitro villus adhesion assay, the pigs (except the B group) were classified as susceptible (s(+)) or nonsusceptible (s(-)) to the intestinal ETEC adhesion. Thus, after the challenge, treatments were B, BChs(-), BChs(+), TrpChs(-), and TrpChs(+). Pigs susceptible to ETEC were 50.0% in the BChs(+) group (3 pigs lost included) and 46.4% in the TrpChs (+) group (1 pig lost included). During the first 4 d after challenge, the challenge reduced ADG (P < 0.05), and this reduction was greater in susceptible pigs (P < 0.05) than nonsusceptible ones. Tryptophan increased ADG and feed intake in susceptible pigs (P < 0.05) from challenge to d 4, but not thereafter. Tryptophan supplementation did not improve the fecal consistency and did not reduce the number of pigs positive for ETEC in feces on d 4 after the challenge. The K88-specific immunoglobulin A activity in blood serum tended to be greater in challenged pigs (P = 0.102) and was not affected by the addition of Trp. Villous height was affected by the addition of Trp and challenge in different ways, depending on the site of small intestine. The need to consider the phenotype for the adhesion of the ETEC in studies with different supply of Trp was clearly evident. When compared with practical weaning standard diets, Trp supplementation allowed susceptible pigs to partially compensate for the effects of ETEC challenge by increasing feed intake and maintaining an adequate BW growth. This is of practical importance for the formulation of diets for pigs selected for lean growth because of the presence of an association between this trait and the susceptibility to the intestinal adhesion of ETEC.

The microbial community in the guts of mammals is often seen as an important potential target in therapeutic and preventive interventions. The aim of the present study was to determine whether enterotoxigenic Escherichia coli (ETEC) F4 infection in young animals might be counteracted by a probiotic treatment with Lactobacillus sobrius DSM 16698. The experiment was conducted in three randomized consecutive replications, each consisting of 16 piglets, and including a control group and an L. sobrius fed group, both experimentally challenged with ETEC. During the entire trial, the animals' health status, body weight, and microbial parameters were monitored periodically. Probiotic supplementation containing L. sobrius significantly reduced the levels of ETEC in the ileum when fed directly to piglets after weaning. In contrast, the number of days when the piglets had an increased faecal water content was significantly higher in the probiotic group. Nevertheless, an improved daily weight gain was also observed in the animals that received probiotic L. sobrius relative to the control fed group. The data indicate that L. sobrius may be effective in the reduction of the E. coli F4 colonization and may improve the weight gain of infected piglets.

Baby formula acidification can be used to reduce diarrhea. Calcium formate is a dietary acidifier frequently used in animal weaning diets; it is also a source of available calcium. Gastric acidification reduces gastrin release and hydrochloric acid (HCl) secretion. To study the medium-term effects on fundic gastric mucosa, we fed weaning pigs control diets or diets supplemented with free or fat-protected calcium formate. We evaluated the following: 1) the number of HCl-secreting parietal cells, by immunohistochemistry using an antibody against H(+)/K(+)-ATPase; 2) the number of enteroendocrine cells immunohistochemically stained with chromogranin A (CGA), somatostatin, and histamine (HIS); and 3) the expression of the H(+)/K(+)-ATPase gene, by real-time RT-PCR in the oxyntic mucosa. Cells co-staining for CGA and HIS were defined as enterochromaffin-like (ECL) cells. Pigs fed calcium formate had fewer parietal cells and a lower expression of the H(+)/K(+)-ATPase gene than the controls (P < 0.05). This reduction did not occur in pigs fed fat-protected calcium formate. Somatostatin immune-reactive cells were also more numerous in pigs fed free calcium formate than in controls (P < 0.05). The number of ECL cells was not affected. Using covariance analysis, the number of parietal cells explained part of the differences in the expression of H(+)/K(+)-ATPase gene (positive correlation, r = 0.385, P < 0.01), and excluded the statistical significance of the diet. In the future, the effects on the oxyntic mucosa should be checked when the diet supplemented with calcium formate is discontinued. Furthermore, a reduction in the number of parietal cells could impair the absorption of vitamin B-12 due to a reduced secretion of the intrinsic factor by these cells.

We investigated whether spray-dried plasma (SDP) improved growth and health of piglets challenged with enterotoxigenic Escherichia coli K88 (ETEC). Forty-eight pigs weaned at 21 d (BW = 4.88 ± 0.43 kg) received one of four diets containing 6% SDP or fish proteins (as-fed basis) either nonmedicated (SDP-NM and FP-NM diets) or medicated with 0 or 250 mg/kg of colistine + 500 mg/kg of amoxycycline (SDP- M and FP-M diets), for 15 d. On d 4, pigs were orally challenged with ETEC. On d 15, eight pigs per dietary group were killed, blood and saliva were collected for analysis of K88 fimbriae-specific immunoglobulin (Ig)- A, and jejunum was removed for villi preparation, histo- logical analysis, and cytokine expression. The presence or absence of K88 receptors (K88 + and K88 − pigs respec- tively) was determined by villous adhesion assay. Ef- fects of protein source on ADG (P = 0.04) and ADFI (P < 0.01), as well of medication on ADFI (P < 0.02), of all pigs were observed. In sacrified pigs, there was an effect of protein source on ADG (P = 0.03) and ADFI (P < 0.001), as well an interaction between medication and presence of K88 receptor (P = 0.02) for feed:gain ratio. Plasma K88 specific IgA were low in all K88 − pigs and higher in K88 + pigs fed FP-NM compared with all the

After challenging piglets with enterotoxigenic Escherichia coli K88 (ETEC), L-tryptophan addition limits the decrease in feed intake and weight gain in ETEC-susceptible subjects. Here we verified whether the susceptibility to ETEC affects the requirement of tryptophan in piglets reared in optimal sanitary conditions and without E. coli K88 challenge. The MUC4 gene was used as a marker to verify the susceptibility, to grouping 36 pigs after weaning, into homozygote for the negative and positive alleles (2 −/− and 2 +/+ pigs per litter). For 21 days pigs were fed using two different diets within each genotype per litter, varied for Trp to Lys ratio (17% or 22%, standardized ileal digestible basis— SID), by a supplementation of 0.5 g/kg L-Trp. In the 1st week feed intake increased with Trp in MUC4+/+ pigs (P

Not nocuous bacteria are important for the maturation and the modulating activity of the gut immune system. However, the humoral immune response against commensal and probiotic bacteria is less documented, particularly in farmhouse animals. Blood serum and saliva were collected in two trials where probiotics, Lactobacillus rhamnosus GG (LbR), well-defined human isolate (Trial A), and a novel and abundant porcine commensal, Lactobacillus sobrius strain 001 T (LbS) (Trial B), were supplemented to weaning pigs. Anti-LbR IgA were present in serum of pigs before treatment with LbR, but also after 1 or 2 weeks in control pigs, notwithstanding the absence of DNA from LbR in colon. Pigs fed or not LbS for 1 or 2 weeks had LbS-specific IgA, in saliva and in serum. Colon contents of control subjects were positive for DNA from LbS. We hypothesized that part of this IgA strain-specific activity is partially related to immune cross-reactivity between different Lactobacillus -species. With the procedure of Shu et al. [Shu, Q., Bird, S.H., Gill, H.S., Rowe, J.B., 1999. Immunological cross-reactivity between the vaccine and other isolates of Streptococcus bovis and Lactobacillus . FEMS Immunol. Med. Microbiol. 26, 153–158], after ELISA test on blood serum or saliva pre-incubated with LbR or LbS, each strain blocked a relevant part of IgA specific for the other. So bacteria with different affinity for the pig present reciprocal crossed immune activity. When probiotics are supplied to weaning pigs, the possible action of already present multi-effective IgA should be considered. The mechanism of IgA induction by certain probiotics needs to be addressed in further studies.

An experiment was conducted to test whether free or fat-protected dietary calcium formate improve the growth and health of weanling pigs which may or may not be susceptible to the intestinal adhesion of enterotoxigenic E. coli (ETEC), when orally challenged with ETEC. Sixty pigs, weaned at 21 days of age (day 0), were divided into three groups (20 pigs each), balanced for number, litter and live weight, and fed the same base diet but three different 30.5 g/kg supplementations: a control (CO – with dicalcium phosphate, calcium sulphate, lard and sepiolite,); a free calcium formate (FF), with this additive, monosodium phosphate and lard; a fat-protected calcium formate (PF), with this additive and monosodium phosphate only. The final diets did not differ for total calcium and phosphorus content. The pigs, individually penned, were orally challenged with 1.5 ml of a 1010 CFU Escherichia coli K88 suspension on day 2 and sacrificed on day 7 or 8. Data were analysed using analysis of variance with a 3-factor design, including diet, block, sensitivity of intestinal villus to ETEC adhesion, and 1st level interactions. The factor diet never interacted with the others. Compared to the control diet, the formate supplementations improved growth (P

64 weaned pigs (24d-old), were fed a control diet or a control plus 1% thymol diet, with or without Salmonella typhimurium challenge. Thymol reduced feed intake but did not affect significantly the final body weight after 29d of trial. Salmonella did not affect the performances. Body temperature was lower in the thymol group before, but also 1d after challenge, when Salmonella increased the values more in unsupplemented pigs. Fecal excretion of S. typhimurium in challenged pigs was not changed by the diet. The thymol group had a higher concentration of immunoglobulin (Ig) in serum before the challenge, but after there was an interaction between diet and challenge. The diet and the challenge did not affect saliva parameters, and ATPase gene expression in the fundic gastric mucosa. Thymol increased the number of parietal cells for 100 μm depth of oxintic gland. In conclusion the important supplementation with thymol reduced the intake, but not performance in the medium period. Some factors of the gut barrier were positively influenced. Salmonella challenge did not reduce significantly the performance of the weaned piglet.

A 50-year-old gardener with a 3-year history of pemphigus vulgaris and steroid-induced diabetes complained of various papulonodular lesions on the left elbow. The lesions had appeared 11 months previously. Skin biopsy showed chronic granulomatous inflammation of the dermis which contained septate hyphae and large spores mainly free. Fragments of the biopsy specimen, cultured on Sabouraud glucose agar without cycloheximide, for 4 days, produced soft white colonies which later turned blackish brown, especially on the underside. On the basis of the microscopic characters of the conidiophores and conidia, Alternaria tenuissima was identified. This phaeohyphomycete has only been isolated previously in two other cases in Italy.

Pentamers of the L1 major capsid protein of human papillomavirus (HPV type 11) were purified after expression in E. coli and analyzed for the kinetics of in vitro capsid self-assembly using multi-angle light scattering (MALS). Pentamers self-assembled into capsid-like structures at a rate that was a function of protein concentration. The kinetics of capsid formation were sigmoidal with a concentration-dependent lag phase, followed by a rapid increase in polymerization. Nucleation size and the rate order of subsequent subunit addition were calculated from the concentration dependence of the extent of capsid formation and the rate of the fast phase, respectively. Assembly was second order with a nucleation size of two pentamers. Thus, we suggest that dimers of pentamers are the nucleus for L1 assembly into capsid-like structures, with rapid sequential addition of single pentamers to the growing shell. Although studied in vitro without accessory factors that may be present in vivo, these data are in contrast with the "five-around-one" assembly nucleus previously proposed for polyomaviruses.

(1986). Vertebrate ecology and ethology. Bollettino di zoologia: Vol. 53, No. sup001, pp. 83-91.

60 suini, svezzati a 21 giorni (d 0), sono stati divisi in 3 gruppi, bilanciati per numero, peso e nidiata ed alimentati con: dieta standard (C), C+1,2% calcio formiato, C+1,2% calcio formiato protetto con grasso. Al giorno 2, i suini sono stati stimolati oralmente con 1,5 ml di una sospensione con 1010 CFU Escherichia coli K88 O148 e sacrificati ai giorni 7 e 8. L’impiego di formiato libero ha migliorato la crescita e l’ingestione, il formiato protetto ho prodotto solo una tendenza al miglioramento. Le due fonti di formiato hanno ridotto l’incidenza di diarrea, l’escrezione fecale di E. coli, le IgA anti E.coli K88 nella saliva e aumentato l’altezza dei villi del piccolo intestino. L’acido formico ha una funzione di promotore di crescita anche in suini stressati con E.coli enterotossigenico. La sua azione è stata però legata principalmente all’effetto sulla persistenza di E.coli totali. Non emerge una particolare convenienza dall’uso di formiato protetto

Importanza della risposta immunitaria nei confronti di commensali residenti e non, per la definizione di strategie d’impiego di probiotici nella dieta del suino in svezzamento. Con l’impiego di probiotici spesso si osserva una maggiore risposta umorale, ma non è documentato se l’animale d’allevamento reagisca anche contro lo stesso probiotico. Sono stati analizzati campioni di sangue e saliva raccolti in due prove di somministrazione di probiotici a suini in svezzamento:A) Lactobacillus rhamnosus GG, non tipico del suino; B) Lactobacillus sobrius 001T, isolato dal suino. Anticorpi probiotico specifici sono stati riscontrati nel siero dei soggetti prima del trattamento (prova A) e nei gruppi di controllo, nonostante l’assenza di DNA ceppo-specifico nel loro contenuto intestinale (prova A e B). Utilizzando un test ELISA su siero o saliva preincubati con uno o l’altro dei lattobacilli, si è visto che ciascuno di questi lega parte delle IgA specifiche per l’altro. Quindi batteri con differente affinità per il suino possono presentare attività immunologia crociata reciproca. Nella definizione dell’impiego di un probiotico nella dieta del suinetto si deve valutare con attenzione la presenza di anticorpi già attivi prima della sua somministrazione.

In a full factorial design (4 diets X challenge, Yes/No), 72 weaning pigs were assigned to one of the diets: Control; experimental diets, obtained with the addition of 2 g/kg free sodium butyrate (fNaB), or 0.6 g/kg fat-protected sodium butyrate (pNaB), or 2 g/kg INVE-NutriAd commercial mixture (Mix, based on 75 g/kg protected butyrate). Oral challenge with Escherichia coli K88 was done on 2/3 of pigs on d 7. Pigs were slaughtered on d 13. The mortality in challenged pigs, tended to be higher in control group (50.0 %) than in the three supplemented groups (23.5%). Growth tended to be increased averagely by the supplements (P = 0.100) after the challenge, that also significantly reduced growth. In general the diet did not affect the fecal shedding of Escherichia coli and Lactobacilli, the K88-specific IgA activity in blood, the morphology of oxyntic mucosa and the expression of H+/K+-ATPase gene. The supplementations tended to increase villous length of jejunum (P = 0.101). On the whole, growth, villous height and surviving rate can be positively affected either when the supplementation is done by free butyrate, by protected butyrate or by the special Inve Nutri-Ad product and these effects are distributed both on pigs infected or not with Escherichia coli K88.