Acanthopanax senticosus extract as a dietary additive enhances the apparent ileal digestibility of amino acids in weaned piglets

The present study was conducted to determine the effects of Acanthopanax senticosus (AS) extract as a dietary additive on serum contents and apparent ileal digestibility (AID) of amino acids (AA) in weaned piglets. In Exp. 1, 60 piglets with an average body weight (BW) of 5.64 kg were randomly assigned into 3 treatment groups who received maize-soybean-based diets supplemented with 0 or 1 g/kg of AS extract or 0.2 g/kg of colistin (n = 20 in each group). Blood samples were randomly collected from 5 piglets per group on days 7, 14 and 28 after the initiation of supplementation to determine the serum contents of free AA. In Exp. 2, 12 barrows with an average initial BW of 7.64 kg were also randomly assigned into 3 dietary treatment groups after being surgically fitted with a simple T-cannula at the terminal ileum. Samples of terminal ileal digesta were collected on day 7 for analysis of the AID of AA. The data showed that the serum contents and AID of most AA in the AS extract-supplemented group gradually increased by 15.3–80.8% and 4.1–30.8%, respectively, as the experiment proceeded in comparison with the colistin-supplemented group and/or control group. In conclusion, these findings indicated that AS extract could enhance the digestion and absorption of AA, which may be a potential mechanism of growth promotion.     

Dietary supplementation with Chinese herbal ultra-fine powder enhances cellular and humoral immunity in early-weaned piglets

This study was conducted to determine effects of dietary supplementation with a Chinese herbal ultra-fine (CHU) powder on cellular and humoral immunity in early-weaned piglets. Sixty piglets weaned at 21 days of age were randomly assigned to one of 3 treatment groups representing supplementation with 0 or 2 g/kg of the powder, or 0.2 g/kg of colistin (an antibiotic) to maize- and soybean meal-based diets (n = 20). On days 7, 14 and 28 after initiation of the treatment, lymphocyte proliferating activity and serum levels of antibodies and cytokines were measured. Results indicated that the powder enhanced (P < 0.05) serum concentrations of IL-2, IL-6 and TNF- on days 7 and 28, and of IgG and IgM on day 28, as well as the proliferating activity of blood lymphocytes in vivo on all days relative to the none additive group. Addition of the powder (7.82 to 31.25 mg/ml) to culture medium in vitro also increased (P < 0.05) the proliferating activity of blood lymphocytes compared with the none additive group. These findings indicate a beneficial effect of the CHU powder on cellular and humoral immune responses in piglets.     

Dietary apple polyphenols supplementation enhances antioxidant capacity and improves lipid metabolism in weaned piglets

Apple polyphenols (APPs) are biologically active flavonoids that have antioxidant, anti‐inflammatory, improving insulin sensitivity, hypocholesterolaemic effect and antiviral properties. This study was conducted to explore effects of dietary APPs supplementation on antioxidant activities and lipid metabolism in weaned piglets. Fifty‐four weaned piglets (half male and female) were randomly divided into three groups with six replicates in each group and three piglets in each repetition. Piglets were fed control diet (basal diet) or a control diet supplemented with 400 mg/kg or 800 mg/kg APPs for 6 weeks. Blood and liver samples were collected to determine biochemical, antioxidant and lipid metabolism parameters. Here we showed that dietary APPs supplementation increased HDL‐C and decreased T‐CHO, TG and LDL‐C concentrations. Dietary APPs supplementation increased antioxidative capacity in serum and CAT activity in liver, and significantly increased the mRNA expressions of CAT, GST and SOD1 in liver. ACC mRNA level and LPL activity were tended to decrease by APPs. HMG‐CoAR, CTP7A1, CD36 and FATP1 mRNA levels were decreased by APPs, while LDL‐R, PGC‐1α, Sirt1 and CPT1b mRNA levels were increased by 400 mg/kg APPs. No alterations in growth performance were found in all treatments. This study firstly provided the evidence that dietary APPs supplementation could enhance systemic antioxidant capacity and improve lipid metabolism in weaned piglets. The mechanism by which APPs improve lipid metabolism might be through regulating hepatic cholesterol metabolism and increasing fatty acid oxidation, and decreasing fatty acid uptake and de novo synthesis.     

Dietary chenodeoxycholic acid improves growth performance and intestinal health by altering serum metabolic profiles and gut bacteria in weaned piglets

Nutritional diarrhea and subsequent performance degradation in weaned piglets are major challenges for the pig industry. Bile acids (BA) can be added to the diet as emulsifiers. This experiment was conducted to investigate the effects of chenodeoxycholic acid (CDCA), a major primary BA, on growth performance, serum metabolic profiles and gut health in weaned piglets. A total of 72 healthy weaned piglets were randomly assigned to the control (CON) and the CDCA groups, which were feed a basal diet and the basal diet supplemented with 200 mg/kg CDCA for 30 d, respectively. Our results demonstrated that CDCA significantly increased final BW and average daily gain (ADG), decreased feed-to-gain (F:G) ratio and tended to reduce diarrhea incidence. In addition, CDCA increased the villus height-to-crypt depth (V:C) ratio, elevated goblet cell numbers and the expression of tight junction proteins, suggesting the enhancement of intestinal barrier function. As an emulsifier, CDCA increased jejunal lipase activity and the mRNA expression of pancreatic lipases. CDCA supplementation also altered the serum metabolic profiles, including increasing the levels of indole 3-acetic acid, N′-formylkynurenine and theobromine that were beneficial for gut health. Moreover, the relative abundance of 2 beneficial gut bacteria, Prevotella9 and PrevotellaceaeTCG-001, were increased, whereas the relative abundance of a harmful bacteria, Dorea, was decreased in the gut of weaned piglets supplemented with CDCA. Importantly, the altered serum metabolic profiles showed a strong correlation with the changed gut bacteria. In conclusion, CDCA improved the growth performance of weaned piglets by improving intestinal morphology and barrier function, and enhancing lipid digestion, accompanied by alterations of serum metabolic profiles, and changes in relative abundance of certain gut bacteria.     

Dietary glutamine supplementation enhances expression of ZO-1 and occludin and promotes intestinal development in Min piglets

Forty-eight Min piglets at 7 days of age were randomly divided into two groups which were fed with 0% and 0.8% glutamine, respectively. Supplementation with 0.8% l-glutamine, the average daily gain (ADG) were significantly increased, and the concentrations of glutamine, glutamate, arginine and citrulline in plasma were significantly improved (P?P?P?P?相似文献   

Expression of gastric ghrelin and H(+)-K(+)-ATPase mRNA in weanling piglets and effect of ghrelin on H(+)-K(+)-ATPase expression and activity in gastric mucosal cells in vitro

This study was designed to investigate the effect of ghrelin on gastric acid secretion in weaning piglets both in vivo and in vitro. Thirty newborn piglets were selected from six litters and on 28, 35 (weaning), 38, 42 and 45d of age, respectively, one piglet from each litter was killed and the mucosal tissue from gastric fundus was collected for detecting ghrelin mRNA as well as H(+)-K(+)-ATPase mRNA expression and activity. Primary cultures of gastric mucosal cells from 5-week-old weaning piglets were challenged with 3x10(-5), 3x10(-4), 3x10(-3), 3x10(-2) and 3x10(-1)nmol/ml h-ghrelin, respectively, for 4h in order to further clarify the effect of ghrelin on gastric H(+)-K(+)-ATPase mRNA expression and activity. Ghrelin mRNA expression in gastric fundus kept stable from 28d to 42d, followed by a sudden increase on 45d, exhibiting a peak that was significantly higher than any other age groups investigated. H(+)-K(+)-ATPase activity and mRNA expression showed similar trends of increase with slightly different timing. H(+)-K(+)-ATPase mRNA expression tended to increase on 42d, while H(+)-K(+)-ATPase activity started to rise from 35d, but neither of them reached significantly higher levels until 45d. In vitro, ghrelin significantly increased H(+)-K(+)-ATPase activity of gastric mucosal cells at 3x10(-4), 3x10(-3), and 3x10(-2)nmol/ml, but augmented H(+)-K(+)-ATPase mRNA expression only at 3x10(-4)nmol/ml. The results indicate that ghrelin mRNA expression is up-regulated 10 days after weaning in the gastric fundus of piglets, coinciding with the increase of H(+)-K(+)-ATPase mRNA expression and activity. Ghrelin acts on gastric mucosal cells to stimulate both mRNA expression and activity of H(+)-K(+)-ATPase in vitro.     

Water extract of Artemisia ordosica enhances antioxidant capability and immune response without affecting growth performance in weanling piglets

The present experiment was conducted to investigate the effects of water extract of Artemisia ordosica (WEAO) on growth performance, antioxidant capability and immune response in weanling piglets. Seventy‐two 28‐day‐old weanling piglets were randomly allocated into four treatments with six replicate pens per treatment and three piglets per pen (n = 18). These four treatment diets were formulated by adding 0, 250, 500 and 750 mg/kg WEAO to the basal diet. The experiment lasted for 28 days. Body weight and feed intake were measured. Blood samples were collected to determine immune and antioxidative parameters. The experimental results showed that WEAO supplementation improved the apparent nutrient digestibility of piglets in a linear or quadratic dose‐dependent manner. In addition, dietary WEAO quadratically increased serum concentrations of cytokines interleukin (IL)‐1, IL‐4, tumour necrosis factor (TNF)‐α, soluble surface antigen CD8 (sCD8), immunoglobulins (Ig)‐A and linearly increased serum concentrations of IL‐2, IL‐6, IgG, IgM. Furthermore, dietary WEAO linearly or quadratically decreased serum concentrations of malondialdehyde but quadratically increased activities of antioxidant enzymes and total antioxidative capacity. These results suggested that WEAO may prove useful as a natural phytogenic feed additive with antioxidative potential and could be incorporated into diets of piglets.     

Dietary glucose oxidase and/or catalase supplementation alleviates intestinal oxidative stress induced by diquat in weaned piglets

This study investigated the effects of dietary exogenous glucose oxidase (GOD) and/or catalase (CAT) on the intestinal antioxidant capacity and barrier function in piglets under oxidative stress. Sixty pigs assigned randomly to five treatment groups—CON: basal diet; DIQ: basal diet; GOD: basal diet + 40-U GOD/kg diet; CAT: basal diet + 50-U CAT/kg diet; and GC: basal diet + 40-U GOD/kg diet + 50-U CAT/kg diet—were analyzed. On Day 14, the CON group was injected with saline, and the others were treated with diquat. The results showed that in diquat-treated piglets, supplementation of dietary GOD and CAT elevated the superoxide dismutase and CAT activities and attenuated the malondialdehyde level in plasma and intestinal mucosa, enhanced the duodenal villus height and villus height/crypt depth ratio, upregulated ZO-1 mRNA level, and attenuated the apoptosis of the epithelial cells and caspase-3 mRNA level in the intestine. Additionally, the supplementation upregulated mRNA expression of the intestinal NF-E2-related factor 2-regulated genes in diquat-treated piglets. However, GOD combined with CAT could not alleviate oxidative damage better than supplementation of CAT or GOD alone under oxidative stress. Overall, the study provides a potential alternative that could relieve the weaning stress in piglets and help formulate antibiotic-free diets.     

Effect of antisecretory drugs on experimentally induced weanling diarrhoea in piglets

In 45 newly-weaned 3 to 4-week-old piglets, diarrhoea was induced by a combined infection with transmissible gastroenteritis (TGE) virus and enterotoxigenic E. coli (ETEC) strains. In untreated control animals this dual inoculation resulted in profuse diarrhoea, vomiting, hypovolaemic shock and death of 77% of the animals within five days of TGE virus inoculation. Antisecretory drugs were administered intramuscularly for three consecutive days after experimental infection. The neurolepticum chlorpromazine, at 2 mg/kg/24 h, resulted in a significant inhibition of diarrhoea and vomiting, and in an increase in weight gain and survival. Sedation and hypothermia, however, were serious side-effects. The ₂ agonist clonidine, at 80 g/kg/12 h, induced a significant antidiarrhoeal effect and a reduction in mortality. The drug, however, provoked decreased activity of ₂-adrenergic excitation and incoordination. The -adrenergic antagonist propranolol, at 0.33 mg/kg/8 h, and the calcium channel blocker verapamil, at 2 mg/kg/8 h, had no beneficial effect on the experimentally induced diarrhoea.     

Prevotella-rich enterotype may benefit gut health in finishing pigs fed diet with a high amylose-to-amylopectin ratio

To investigate the influence of baseline enterotypes and dietary starch type on the concentration of short-chain fatty acids (SCFA), numbers of butyrate producing bacteria and the expression of genes related to intestinal barrier and inflammatory response in the colon of finishing pigs, a 60-d in vivo trial was conducted. A 2-wk pre-trial with 102 crossbred (Duroc × [Landrace × Yorkshire]) finishing barrows (90 d old) was conducted to screen enterotypes. Then, a total of 32 pigs (87.40 ± 2.76 kg) with high (HPBR, ≥ 14) and low (LPBR, ≤ 2) Prevotella-to-Bacteroides ratios (PBR) in equal measure were selected and randomly divided into 4 groups with 8 replicates per group and 1 pig per replicate. The trial was designed following a 2 (PBR) × 2 (amylose-to-amylopectin ratio, AMR) factorial arrangement. Pigs with different PBR were fed diets based on corn-soybean meal with high AMR (HAMR, 1.24) or low AMR (LAMR, 0.23), respectively. Results showed that neither PBR nor AMR influenced the growth performance of pigs. HPBR pigs fed HAMR diet had a higher number of colonic Clostridium cluster XIVa and higher gene expression of butyrate kinase compared to the LPBR pigs (P < 0.05). The HPBR pigs fed HAMR diets also had increased colonic concentrations of total SCFA and propionate compared to the LPBR pigs (P < 0.05). Comparing with other pigs, HPBR pigs fed HAMR diets showed a lower (P < 0.05) expression of histone deacetylases (HDAC) gene and higher (P < 0.05) expression of G protein-coupled receptor 43 gene (GPR 43) in the colonic mucosa. The interaction (P < 0.05) of HPBR and HAMR was also found to decrease the gene expression of interleukin (IL)-6, IL-12, IL-1β and tumor necrosis factor-α (TNF-α) in colonic mucosa. These findings show that HAMR diet increased the abundance and activity of butyrate-producing bacteria and the concentration and absorption of SCFA, which may be associated with the decreased gene expression of inflammatory cytokines in the colonic mucosa of pigs with Prevotella-rich enterotype. All these alterations are likely to have a positive effect on the intestinal health of finishing pigs.     

Molecular screening and risk factors of enterotoxigenic Escherichia coli and Salmonella spp. in diarrheic neonatal calves in Egypt

The aim of the present study was to carry out molecular epidemiological investigation on enterotoxigenic Escherichia coli (ETEC) K99 and Salmonella spp. in diarrheic neonatal calves. Fecal samples were obtained from 220 diarrheic calves at 9 farms related to four governorates in central and northern Egypt. E. coli and Salmonella spp. isolates were examined for E. coli K99 and Salmonella spp. using PCR. ETEC K99 was recovered from 20 (10.36 %) out of 193 isolates, whereas Salmonella spp. was recovered from nine calves (4.09%).Multivariable logistic regression was used to evaluate the risk factors associated with both infections. ETEC K99 was significantly affected by age (P < 0.01; OR: 1.812; CI 95%: 0.566–1.769), colostrum feeding practice (P < 0.01; OR: 5.525; CI 95%: 2.025–15.076), rotavirus infection (P < 0.001; OR: 2.220; CI 95%: 0.273–1.251), vaccination of pregnant dams with combined vaccine against rotavirus, coronavirus and E. coli (K99) (P < 0.001; OR: 4.753; CI 95%: 2.124–10.641), and vitamin E and selenium administration to the pregnant dam (P < 0.01; OR: 3.933; CI 95%: 0.703–1.248).Infection with Salmonella spp. was found to be significantly affected by the animal age (P < 0.05; OR: 0.376; CI 95%: 0.511–1.369), Hygiene (P < 0.05; OR: 0.628; CI 95%: 1.729–5.612), and region (P < 0. 01; OR: 0.970; CI 95%: 0.841–1.624).The results of the present study indicate the importance of PCR as rapid, effective and reliable tool for screening of ETEC and Salmonella spp. when confronted with cases of undifferentiated calf diarrhea. Moreover, identification of the risk factors associated with the spreading of bacteria causing diarrhea may be helpful for construction of suitable methods for prevention and control.     

Anaplasma marginale infection in a Japanese Black cow 13 years after eradication of Rhipicephalus (Boophilus) microplus in Okinawa, Japan

In October 2007, a 15-year-old Japanese Black cow on Ishigaki Island, Okinawa, Japan, was diagnosed with Anaplasma marginale infection based on clinical symptoms, blood examination, smear observation, 16S rRNA and groEL gene sequence analysis, and the result of a CF test. The cow was introduced into the farm from mainland Japan as a calf in 1993, one year before the eradication of Rhipicephalus (Boophilus) microplus, the main vector of A. marginale in Okinawa Prefecture. It is possible that the cow was first infected with A. marginale as a calf in Ishigaki Island and had been persistently infected since then. This is the first reported clinical case of A. marginale infection of cattle since the eradication of R. microplus in Okinawa Prefecture. Additional analysis of major surface protein 1α amino acid sequences revealed that the A. marginale Okinawa strain presented four new repeat forms which were not seen in other strains. This indicates that the Okinawa strain may be a unique geographical variant of A. marginale.     

Natural infection by Cryptosporidium sp., Giardia sp. and Eimeria leuckarti in three groups of equines with different handlings in Rio de Janeiro, Brazil

To detect Cryptosporidium sp., Giardia sp. and Eimeria leuckarti in horses, fecal samples were collected from three different handling horse groups from the state of Rio de Janeiro, Brazil. Group A was composed of “Mangalarga Marchador” pure breed horses, Group B was formed by horses of a Military Corporation and Group C by stray horses captured by the Center of Zoonosis Control Paulo Dacorso Filho. A total of 396 fecal samples were collected, 212 samples from Group A, 154 samples from Group B and 30 from Group C. The material was submitted to the centrifugation - flotation technique and staining by the safranin-methylene blue technique and analyzed. Oocysts of Cryptosporidium sp. were identified in 0.75% of the samples (n = 3); cysts of Giardia sp. in 0.5% (n = 2) and oocysts of E. leuckarti in 0.5% (n = 2). One case of E. leuckarti in group A and one of Cryptosporidium sp. in group B were observed. In group C were observed two cases of Cryptosporidium, two of Giardia and one of E. leuckarti,. Horses of group C were more parasitized by the three protozoans than animals from the other groups (p < 0.01). It was possible to verify that factors related to the animals, like host individual susceptibility and sanitary factors may influence the occurrence of natural infections by gastrointestinal protozoans, although the age did not have influence. This study reports, for the first time, the occurrence of Cryptosporidium sp., Giardia sp. and E. leuckarti in equines of the State of Rio de Janeiro.     

Effect of Momordica charantia L. in the resistance to aminoglycosides in methicilin-resistant Staphylococcus aureus

In this study the ethanol extract (EEMC) of Momordica charantia L. (Cucurbitaceae) was tested for its modifying antibiotic activity against a MRSA strain. The growth of an MRSA (SA358) in the absence and presence of aminoglycosides was evaluated. A potentiating effect between this extract and all aminoglycosides was demonstrated. Similarly, the same effect was shown by chlorpromazine on kanamycin, gentamicin and neomycin, indicating the involvement of an efflux system in the resistance to these aminoglycosides. Extracts from M. charantia could be used as a source of plant-derived natural products with resistance-modifying activity. This is the first report about the modifying antibiotic activity of M. charantia, constituting a new weapon against multi-resistant bacteria such as MRSA.     

Seroprevalence of antibodies to Encephalitozoon cuniculi and Encephalitozoon intestinalis in humans and animals

The presence of antibodies against Encephalitozoon cuniculi (E. cuniculi) and Encephalitozoon intestinalis (E. intestinalis) was examined in 215 samples from humans and in 488 samples from five different species of domestic and companion animals in Slovakia. The 215 human samples and samples from 90 swine, 123 non-infected cattle (cattle), 24 cattle infected with bovine leukosis virus (BLV-positive cattle), 140 sheep and 111 dogs were examined by the enzyme-linked immunosorbent assay (ELISA). Samples with serum titres 1:200 or higher were considered as positive. Specific anti-E. cuniculi antibodies were found in humans (0.9%), swine (52%), cattle (2%), sheep (9%) and dogs (15%) except for the BLV-positive cattle at the titre of 1:200. The titre of 1:400 was detected only in humans (0.5%). The presence of specific anti-E. intestinalis antibodies at the titre of 1:200 was confirmed in humans (6%), swine (51%), cattle (11%), BLV-positive cattle (13%) and dogs (6%) but not in sheep. The anti-E. intestinalis antibodies reached the 1:400 in humans (1%), swine (4%) and BLV-positive cattle (17%). The presence of specific anti-E. intestinalis antibodies at the titre of 1:600 was observed only in one swine (1%). Significant differences were observed in animals at titres 1:200 and 1:400 (chi-squared test: p < 0.0001) for both pathogens and in humans only for E. cuniculi at the titre of 1:400 (chi-squared test: p < 0.0075).     

Protection against Toxoplasma gondii brain cyst formation in mice immunized with Toxoplasma gondii cytoskeleton proteins and Lactobacillus casei as adjuvant

The aim of the study was to evaluate the protection generated in mice against Toxoplasma gondii brain cyst burden by vaccination with T. gondii cytoskeleton proteins using Lactobacillus casei as adjuvant. One hundred and sixty-eight NIH mice were randomly allocated into eight groups of 21 mice each. Animals were immunized as follows: in group 1 with Toxoplasma lysate antigen (TLA) in Freund's modified adjuvant, containing L. casei (FMA), in group 2 with Toxoplasma cytoskeleton proteins (TCPs) in FMA, in group 3 with FMA, in group 4 with phosphate buffered saline (PBS), in group 5 with L. casei dead by heath (Lc), in group 6 with Freund's complete adjuvant (FCA), in group 7 with TLA in FCA, and in group 8 with TCP in FCA. Mean brain cyst burden (±S.E.M.) was assessed in mice 8 weeks after challenge with T. gondii Me49 strain (20 cysts per mouse). The percentages of reduction in cyst burden per brain (P < 0.01) as compared with the group 4 (control: mean 3181 ± 97.5) were 77.25% (724 ± 98) in group 1, 88.02% (381 ± 97.5) in group 2, 38.92% (1943 ± 130.3) in group 3, 44.31% (1771.4 ± 102) in group 5, 59.28% (1295.2 ± 99.1) in group 7 and 55.69% (1409.5 ± 89.9) in group 8. In order of importance, the best protection was obtained in groups 2, 1, 7, 8, 5 and 3. Noticeably the mice inoculated with L. casei alone showed a significant reduction in T. gondii brain cysts (P < 0.01), while those animals treated with FCA alone did not. Additionally, IgM anti-T. gondii antibody levels, as determined by ELISA 2 weeks after challenge, were highest in group 2 (P < 0.01) than in the other seven groups. Results suggest that T. gondii cytoskeleton proteins with L. casei as adjuvant constitute a good anti-toxoplasmosis vaccine candidate.     

Synergistic and antagonistic hemolytic activities of bacteria of genus Arcanobacterium and CAMP-like hemolysis of Arcanobacterium phocae and Arcanobacterium haemolyticum with Psychrobacter phenylpyruvicus

A total of 57 bacteria representing eight species of genus Arcanobacterium (A.) were investigated for hemolytic properties on blood agar containing sheep and rabbit blood and for CAMP-like reactions. An enhanced hemolysis on blood agar containing rabbit blood compared to sheep blood could be observed for A. haemolyticum, less pronounced for A. hippocoleae and A. pluranimalium.A synergistic hemolytic reaction with staphylococcal β-hemolysin appeared to be constantly visible for A. hippocoleae, A. pluranimalium and A. pyogenes, with Streptococcus agalactiae for A. phocae and A. haemolyticum, with Rhodococcus equi for A. phocae, A. haemolyticum, A. pluranimalium and A. pyogenes and with A. haemolyticum for A. hippocoleae, A. pluranimalium and A. pyogenes, respectively. A reverse CAMP-reaction in the zone of staphylococcal β-hemolysin could be observed for A. phocae and A. haemolyticum. In addition, a novel CAMP-like reaction could be noted between Psychrobacter phenylpyruvicus, identified by 16S rDNA sequencing, and A. phocae and A. haemolyticum. These synergistic or antagonistic hemolytic properties could possibly be used as additional criteria for identification of bacteria of genus Arcanobacterium.     

Role of nitric oxide production in dairy cows naturally infected with Mycobacterium avium subsp. paratuberculosis

Nitric oxide (NO) is a crucial mediator in host defense and is one of the major killing mechanisms within macrophages. Its induction is highly affected by the types of cytokines and the infectious agents present. In the current study, NO production was evaluated after in vitro infection of unfractionated peripheral blood mononuclear cells (PBMCs) with Mycobacterium avium subsp. paratuberculosis (MAP) after 8 h, 3 and 6 days of culture for cows in different stages of disease. In addition, the effects of in vitro exposure to inhibitory cytokines such as interleukin-10 (IL-10) and transforming growth factor β (TGF-β) as well as the pro-inflammatory cytokine IFN-γ were correlated with the level of NO production. Nitric oxide production was consistently higher in cell cultures from subclinically infected animals at all time points. An upregulation of NO production was demonstrated in unfractionated cell cultures from healthy control cows after exposure to MAP infection as compared to noninfected cell cultures. A similar increase in NO due to the addition of MAP to cell cultures was also noted for clinically infected cows. NO level among subclinically infected cattle was greater at all time points tested and was further boosted with the combination of both in vitro MAP infection and IFN-γ stimulation. Alternatively, nonspecific stimulation with LPS from Escherichia coli O111:B4-W resulted in an upregulation of NO production in all infected groups at 3 and 6 days after in vitro infection. Finally, the in vitro exposure to inhibitory cytokines such as IL-10 and TGF-β prior to MAP infection or LPS stimulation resulted in the downregulation of this inflammatory mediator (NO) in all experimental groups at all time points. In summary, a higher level of NO production was associated with cows in the subclinical stage of MAP infection. As well, the results demonstrated an increase in NO production upon infection with MAP and in the presence of exogenous IFN-γ. Finally, the results suggest an important role of IL-10 and TGF-β on the profile of NO production which may explain the low NO production in MAP clinically infected cows.     

Effects of Saccharomyces cerevisiae on ruminal pH and microbial fermentation in dairy cows: Yeast supplementation on rumen fermentation

An experiment was conducted with eight ruminally-cannulated cows using a crossover design with 2 periods to determine the effects of yeast supplementation on rumen fermentation. Holstein dairy cows in late lactation were either supplemented with 0.5 g/hd/d of Saccharomyces cerevisiae, an active dry yeast (CNCM-1077, Levucell SC20 (r) SC, Lallemand Animal Nutrition) or not supplemented (control). A basal diet consisting of 60% forage and 40% concentrate (DM basis) was fed once daily to both groups of cows throughout the entire experiment. Ruminal pH was measured continuously every 22 min using a pH probe that was placed in the ventral rumen sac for 6 consecutive days. Volatile fatty acid and ammonia N concentrations in the rumen were measured on days 5 or 6 of the 12-d period for each cow and DM intake was monitored throughout the experiment. Data were analyzed using a mixed-effects model with repeated measures. There were no differences in dry matter intake between treatments. Mean ruminal pH was greater (P < 0.05) when yeast was supplemented (6.53 ± 0.07) compared with the control (6.32 ± 0.07). Average maximum and minimum ruminal pH were also greater (P < 0.05) when yeast was supplemented (7.01 ± 0.09 and 5.97 ± 0.08, respectively) compared with the control (6.80 ± 0.09 and 5.69 ± 0.09, respectively). Time spent under the subacute acidosis threshold, pH less than 5.6, was lower (P < 0.05) with yeast supplementation compared with control cows. No difference was observed for ruminal ammonia N concentrations (mean = 14.0 ± 1.2 mg/dL) between treatments. Total VFA concentration (mM) in the rumen tended to be lower (P = 0.10) in the yeast-supplemented cows (107.3 ± 6.35) than in the control cows (122.4 ± 6.35), which could be related to the greater pH observed with yeast supplementation. Supplementing dairy cows with active dry yeast in the current experiment increased the mean, minimum and maximum ruminal pH; decreased time spent in subacute rumen acidosis, and tended to decrease total VFA concentration in the rumen compared with control cows.