Design and validation of an orally administrated active L. fermentum-L. acidophilus probiotic formulation using colorectal cancer Apc Min/+ mouse model

Probiotics have been shown to have beneficial properties in attenuating the risk of colorectal cancer (CRC) development. However, functional evidence to support such effects for some probiotic bacteria are relatively unknown. Here, we document a significant antioxidant, anti-proliferative and pro-apoptotic activities of Lactobacillus acidophilus ATCC 314 and Lactobacillus fermentum NCIMB 5221 on CRC cells, particularly when used in combination (La-Lf). Furthermore, a superior synergistic activity on the inhibition of tumor growth and modulation of cell proliferation and epithelial markers in the Apc ^Min/+ CRC mouse model was explored, based on the expression levels of Ki-67, E-cadherin, β-catenin, and cleaved caspase-3 (CC3) proteins. The anti-cancer activity of La-Lf co-culture was significantly enhanced in vitro with significant reduced proliferation (38.8 ± 6.9 %, P = 0.009) and increased apoptosis (413 RUL, P < 0.001) towards cancer cells, as well as significant protection of normal colon cell growth from toxic treatment (18.6 ± 9.8 %, P = 0.001). La-Lf formulation (10¹⁰cfu/animal/day) altered aspects of intestinal tumorigenesis by significantly reducing intestinal tumor multiplicity (1.7-fold, P = 0.016) and downregulating cellular proliferation markers, including β-catenin (P = 0.041) and Ki-67 (P = 0.008). In conclusion, La-Lf showed greater protection against intestinal tumorigenesis supporting a potential use as a biotherapeutic for the prevention of CRC.

     

Influence of <Emphasis Type="Italic">Lactobacillus fermentum</Emphasis> I5007 on the intestinal and systemic immune responses of healthy and <Emphasis Type="Italic">E. coli</Emphasis> challenged piglets

The effect of feeding Lactobacillus fermentum I5007 on the immune system of weaned pigs with or without E. coli challenge was determined. Twenty-four weaned barrows (6.07 ± 0.63 kg BW) were randomly assigned to one of four treatments (N = 6) in a factorial design experiment. The first two treatments consisted of healthy piglets with half of the pigs receiving no treatment while the other half was orally administered with L. fermentum I5007 (10⁸ CFU/ml) at a daily dose of 20 ml. Pigs in the second two treatments were challenged on the first day with 20 ml of E. coli K88ac (10⁸ CFU/ml). Half of these pigs were not treated while the remaining pigs were treated with 20 ml of L. fermentum I5007 (10⁸ CFU/ml). Peripheral blood lymphocytes subsets were determined using flow cytometry. The intestinal mucosal immunity of the pigs was monitored by real time polymerase chain reaction. The cytokine content of the pig’s serum was also analyzed. Oral administration of L. fermentum I5007 increased blood CD4⁺ lymphocyte subset percentage as well as tumor necrosis factor-α and interferon-γ expression in the ileum. Pigs challenged with E. coli had elevated jejunal tumor necrosis factor-α while interferon-γ expression was increased throughout the small intestine. There was no difference in the concentration of the cytokines interleukin-2, interleukin-6, tumor necrosis factor-α and interferon-γ in the serum. CD8⁺ and CD4⁺/CD8⁺ in peripheral blood were not affected by treatment. In conclusion, L. fermentum I5007 can enhance T cell differentiation and induce ileum cytokine expression suggesting that this probiotic strain could modulate immune function in piglets.     

Effect of Lactobacillus fermentum alone, and in combination with zinc(II) propionate on Salmonella enterica serovar Düsseldorf in Japanese quails

The influence of zinc(II) propionate on the efficacy of Lactobacillus fermentum against Salmonella enterica serovar Düsseldorf was tested in Japanese quails. Twenty one 3-day old Japanese quails were divided into 3 groups each consisting of 7 birds and inoculated orally: (i) group A (control) with Rogosa broth; (ii) group B with rifampicin-resistant L. fermentum (2.5 × 10⁷ CFU/animal); and (iii) group C with rifampicin-resistant L. fermentum cultured in Rogosa broth supplemented with zinc(II) propionate (1 g Zn²⁺ /L of broth; i.e. 2.5 × 10⁷ CFU/animal and 0.1 mg Zn²⁺ /animal). After 16 hours all birds were infected with a single dose of S. enterica serovar Düsseldorf (5.8 × 10⁷ CFU /animal). During the next 6 days the chicks received the same inoculations as they had received earlier through the medium of drinking water. The viable counts of Salmonella, rifampicin-resistant lactobacilli and total aerobes in the feces and cecal content, zinc concentration in the cecal content and growth performance were evaluated. L. fermentum alone, and in combination with Zn(II) propionate significantly reduced shedding of Salmonella in the feces and also the amount of salmonellae present in cecal content as compared to control. The numbers of Salmonella in group C in all collections were lower than in group B, but a significant difference (p < 0.05) was noted only 24 h after infection. The viable counts of rifampicin-resistant lactobacilli were similar in groups B and C and numbers of total aerobes were reduced in these groups compared with control. L. fermentum and its combination with zinc(II) propionate increased daily weight gains in the chicks in comparison with control. Zinc concentration in the control and C group was 34.9 ± 6.2 mg · kg⁻¹ and 676.3 ± 106.6 mg · kg⁻¹ of cecal content, respectively. Presented at the Second Probiotic Conference, Košice, 15–19 September 2004, Slovakia.     

Efficacy of CIM 1166, a combination of compounds derived from Mentha spp. in alleviating experimental vulvovaginal candidiasis in mice

Candida albicans is yeast that is most often associated with serious fungal infections and can cause fungal diseases in immuno-compromised patients especially patients suffering from AIDS, cancer and cases of organ transplant. Amongst women, candidal vaginitis is predominantly caused by strains of Candida albicans and also remains to be a common problem in immuno-competent or healthy women. A study was undertaken to assess the efficacy of a compound CIM 1166 obtained from plant source which was found to possess promising antimicrobial property under in vitro conditions especially against C. albicans. Taking the lead further, a small animal model utilizing aged Swiss albino females that had parturated at least three times were taken up for model development. Infection (7 × 10⁶ cfu/ml) was instilled into the vagina in a volume of 20 μl for 3 days. Vaginal washings were aseptically collected on day 4th to confirm the establishment of infection following which the treatment was started which continued for the next 5 days through vaginal route. Vaginal washings were collected on 6th day and the colony forming units were enumerated on chloramphenicol incorporated SDA plates. The results indicated that there was a significant decrease in the colony forming units in vaginal washings (8.0 × 10² cfu/ml) of the treated animals as compared to blank control group (6.0 × 10⁴ cfu/ml). The positive control group administered with clotrimazole also showed a recovery from infection with a fungal load of 8.78 × 10² cfu/ml. The study proves the efficacy of CIM 1166 in curing vaginal candidiasis in mice, which can be taken up for formulation development and further studies.     

Administration of Lactobacillus fermentum CRL1446 increases intestinal feruloyl esterase activity in mice

Aims: To evaluate the effect of oral administration of Lactobacillus fermentum CRL1446 on the intestinal feruloyl esterase (FE) activity and oxidative status of mice. Methods and Results: Adult Swiss albino mice received Lact. fermentum CRL1446 at the doses 10⁷ and 10⁹ cells per day per mouse for 2, 5, 7 and 10 days. Intestinal FE activity, intestinal microbiota counts, plasmatic thiobarbituric acid‐reactive substances (TBARS) percentage and glutathione reductase (GR) activity were determined. Mice that received Lact. fermentum CRL1446 at the dose 10⁷ cells per day for 7 days showed a twofold increase in total intestinal FE activity, compared to the nontreated group. In large intestine content, FE activity increased up to 6·4 times. No major quantitative changes in colonic microbiota were observed in treated animals. Administration of this strain produced an approx. 30–40% decrease in the basal levels of plasmatic TBARS and an approx. twofold increase in GR activity from day 5 of feeding with both doses. Conclusions: Oral administration of Lact. fermentum CRL1446 to mice increases total intestinal FE activity, decreases the basal percentage of plasmatic lipoperoxides and increases GR activity. Significance and Impact of the Study: Lactobacillus fermentum CRL1446 could be orally administered as a dietary supplement or functional food for increasing the intestinal FE activity to enhance the bioavailability of ferulic acid, thus improving oxidative status.     

Microbiological condition and shelf life of meat from hunted game birds

Hunted game birds (eight partridges, nine wood pigeons, 25 quails, 16 chilled and 16 frozen–thawed pheasants) were processed according to “Good Manufacturing Practice” rules. Microflora of skin, intestinal content and meat cuts (breast and thigh, both fresh and stored in vacuum package) was analysed. Listeria monocytogenes, Salmonella sp. and Campylobacter sp. were not recovered from any sample. Log microbial numbers on skin or in intestines were not significantly related to those on meat cuts. With the exception of pigeons, microbial numbers of the two meat cuts did not differ significantly (p > 0.05), and no significant increase in microbial numbers in vacuum-stored meat was found; the same applied to frozen–thawed compared to chilled pheasants. On meat, average total viable counts were <4.00 log cfu/cm² with a maximum of 6.48 log cfu/cm². Median Escherichia coli numbers were <2.00 log cfu/cm², maximum was 4.48 log cfu/cm². Meat cuts obtained from partridges, quails and pheasants demonstrated a shelf life of 1 week, provided they were kept vacuum-packaged at 0°C to 1°C.     

Lactobacillus gasseri LF221 and K7 — from isolation to application

The article presents research findings on two human strains with probiotic activity. On the basis of API 50 CHL fermentation pattern, PCR by species-specific primers and sequencing of the V2–V3 region of 16S rRNA both strains designated as LF221 and K7 were identified as members of the Lactobacillus gasseri species. Two LF221 bacteriocins, acidocin LF221 A and B were purified and sequenced. They were classified as members of the two-component class II bacteriocins. Among basic probiotic properties, the survival under conditions in gastro-intestinal tract, ability to adhere to cultured intestinal enterocytes and pig’s mucosa and stimulation of the immune response were demonstrated. In in vivo study of 24 weaned piglets, the survival rate of K7 Rif^r and LF221 Rif^r was quantified by selective enumeration on MRS agar with rifampicin. The survival of both strains was good (2.9 × 10⁵ cfu of K7 Rif^r /g faeces; 4.8 × 10⁵ cfu of LF221 Rif^r /g) and the LF221 Rif^r /K7 Rif^r viable cells were found either in the mucosa of duodenum, jejunum or in the ileum. The possible effect of K7 to inhibit adhesion of E. coli O8:K88 to enterocytes was studied on Caco-2 cultured cells, on tissue obtained from small intestines of pigs and in vivo on gnotobiotic piglets. Lactobacilli were found to be effective in reducing E. coli adhesion to enterocytes in Caco-2 model, but not on mucosa of pig’s jejunum under ex vivo conditions. Competitive exclusion, production of organic acids and stimulation of immune response, were involved in inhibition of E. coli by K7 strain in gnotobiotic piglets. Any inflammatory change in intestines of piglets treated with K7 was observed, which confirmed its safe use. Among the technological parameters the survival and activity of the strains during cheese-making are presented. Presented at the Second Probiotic Conference, Košice, 15–19 September 2004, Slovakia.     

Multi-species probiotics improve growth,intestinal microbiota and morphology of Indian major carp mrigal Cirrhinus cirrhosus

This study aimed to examine the effects of multi-species probiotic on growth, hematological status, intestinal microbes, and intestinal morphology of mrigal (Cirrhinus cirrhosus). The mrigal fries (average weight 0.51 g) were reared for 60 days by supplementing multi-species probiotics containing Bacillus spp. (1 × 10⁹ cfu/mL) and Lactobacillus spp. (1 × 10¹¹ cfu/mL) in the raising water at doses of 0 (control), 0.5, and 1.0 mL/L. The results indicated that fish reared with multi-species probiotics showed significantly higher growth performance and feed efficiency where the survival rate was similar in all cases. Accordingly, significant higher red blood cell (RBC) and white blood cell (WBC) were counted from the fish reared with multi-species probiotic. There was a considerable difference in bacterial microbiota between the experimental and control group. Multi-species probiotics significantly enhanced the length, width, and villus area. Several immune response indicators like fattening of intestinal mucosal fold, width of lamina propria, the width of enterocytes, and abundance of goblet cells were also increased significantly in fish that received multi-species probiotics. This study revealed that multi-species probiotics can significantly contribute to the growth of mrigal through upgrading intestinal microbiota and morphology, which can be suggested as an eco-friendly growth stimulator in mrigal farming.     

Effect of bovine lactoferrin in <Emphasis Type="Italic">Salmonella</Emphasis> ser. Typhimurium infection in mice

Lactoferrin (LF) has in vitro antimicrobial activity against Gram-negative bacteria. Salmonella enterica subsp. enterica serovar Typhimurium causes systemic infection and acute diarrhea in humans, mainly in children younger than 2 years of age. The aim of the study was to determine the in vivo effect of bovine LF in Salmonella ser. Typhimurium infection in mice. 58 BALB/c mice were employed. Two hours before the infection with 300 μl of 10⁷ CFU of Salmonella ser. Typhimurium, 29 mice received LF (2 mg) and 29 placebo (buffer). After the infection, the mice received LF (10 mg/ml) ad libitum or buffer, respectively, for 7 days. Mortality, weight and clinical signs (piloerection, hunched position and reduced movement) were monitored daily. The degree of inflammation and necrosis in the intestine, liver, spleen and brain were studied with a blinded observer. The mortality in the control group (8/29) was higher than in the LF group (1/29) (Kapplan Meier P < 0.05). From the third day post-infection the control group were significantly more symptomatic (P < 0.05). The blood culture for Salmonella spp. was positive for all mice studied in the control group (17/17), but positive in the LF group in only 6/17 animals (P < 0.05). In the LF group, the pathologic studies show less inflammation and focal necrosis in the four organs studied, with the greatest difference found in the intestine. Bovine LF protects against Salmonella ser. Typhimurium infection in mice, reducing the severity, mortality and the degree of inflammation of this infection.     

Effects of Dietary Supplementation of Recombinant Plectasin on Growth Performance,Intestinal Health and Innate Immunity Response in Broilers

The present study was conducted to evaluate the effects of dietary supplementation of recombinant plectasin (Ple) on the growth performance, intestinal health, and serum immune parameters in broilers. A total of 288 1-day-old male broilers (Arbor Acres) were randomly allotted to four dietary treatments including the basal diet (NC) and basal diet supplemented with 10 mg enramycin/kg (PC), 100 mg Ple/kg (LPle), and 200 mg Ple/kg (HPle) diets. The results indicated Ple increased (P < 0.01) average daily gain and decreased (P ≤ 0.02) feed to gain ratio of broilers. In addition, the supplementation of Ple in the diets increased (P ≤ 0.01) duodenal lipase (day 21) and trypsin (day 42) activities compared with the NC group. Similar as the supplementation of enramycin, Ple also increased villus height and decreased crypt depth in jejunum (day 21), and thus the villus height to crypt depth ratio (P < 0.01) was increased compared to the NC group on day 42. The serum immunoglobulin M (days 21 and 42), immunoglobulin G (day 42), complement 3 (day 21), and complement 4 (days 21 and 42) were significantly increased (P ≤ 0.02) due to the supplementation of Ple and enramycin, while the concentration of malondialdehyde in jejunum was decreased (P < 0.01) in PC, LPle, and HPle groups on day 21 compared with those in the NC group. Furthermore, Ple reduced (P < 0.01) Escherichia coli and total aerobic bacteria population in ileum and cecum of birds on days 21 and 42. These results indicate that the recombinant plectasin has beneficial effects on growth performance, intestinal health, and innate immunity in broilers.

     

Effects of dietary zearalenone on the serum biochemistry,hepatic and intestinal histology,and intestinal microbiota of juvenile Dabry′s sturgeon (Acipenser dabryanus)

Zearalenone (ZEN) is a common contaminant in animal feed, but the adverse effects of ZEN on the intestinal microbiota of fish have not yet been investigated. To reveal the effects of ZEN on serum biochemistry, hepatic and intestinal histology, and intestinal microbiota of fish, Dabry′s sturgeon (Acipenser dabryanus) received feed containing 1030 μg kg⁻¹ ZEN (ZEN-treated group) for 7 weeks and were compared to a control feed that have not been fortified with ZEN (control group). The results showed that dietary supplementation with ZEN did not significantly affect the serum contents of alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, high-density lipoprotein cholesterol (HDLC) and estradiol (E₂; p > .05). The serum contents of total protein, albumin, triglycerides, total cholesterol and low-density lipoprotein cholesterol in the ZEN-treated group were significant lower than the control group (p < .05). The serum contents of glucose in the ZEN-treated group was significant higher than the control group (p < .05). Intestinal histology was normal in the ZEN-treated group with comparsion to the control group. Compared to the control group, the appearance of nuclear pyknosis and vacuoles in the hepatic cell in the ZEN-treated group. The α-diversity index (Chao 1, Faith pd and Shannon diversity index) significant increased in the ZEN-treated group compared to the control group (p < .05). Simpson diversity index was not affected by the dietary ZEN-treated (p > .05). Principal coordinates analysis (PCA) showed different clustering of prokaryotic communities in the ZEN-treated group compared to the control group. Further Anosim (Analysis of similarities) found that significnat differences in species community structure composition (R > .44) between the control group and the ZEN-treated group (p < .05). The number of operational taxonomic units (OTUs) was decreased after Dabry's sturgeon were fed diets supplemented with ZEN. At the genus level, the differences in the relative abundances of the first 20 main microbiota genera were considerable. In summary, this study suggests that dietary containing 1030 μg kg⁻¹ ZEN may be interfere with physiological parameters, and also affect the intestinal microbiota α-diversity, the numbers of OTUs and the microbiota compostion at the genus level of the juvenile Dabry′s sturgeon.     

Evaluation of Probiotic <Emphasis Type="Italic">Lactobacillus fermentum</Emphasis> CCM 7421 Administration with Alginite in Dogs

There are growing efforts to find applications for various naturally occurring organo-mineral rocks. They have so far been preferentially used in agriculture and forestry; however, medicine and nutrition may also be interesting areas for their application. This study investigates the effects of dietary supplementation with canine-derived probiotic strain Lactobacillus fermentum CCM 7421 in combination with alginite in dogs. Alginite is a loam-like material of volcanic origin composed of clay minerals and fossilised unicellular algae. The effects of these additives on faecal microbiota, faecal characteristics, short-chain fatty acid profile, haematology, serum biochemistry and cellular immunity parameters were monitored. Forty dogs were randomly divided into four treatment groups: control group (C), alginite-supplemented group (A; 1% diet), probiotic group (LF; L. fermentum CCM 7421 at a dose of 10⁹ cfu/day/dog) and combined group (A + LF group); 10 dogs in each group. The experiment lasted for 35 days with a 14-day treatment period (sample collection at days 0, 7, 14 and 35). The results of this straightforward experiment showed beneficial effects in the combined A + LF group. In detail, a decrease in faecal coliforms and clostridia and an increase in lactic acid bacteria, haemoglobin and serum magnesium levels compared to baseline were observed in the A + LF group (P?<?0.05). In contrast, sole application of alginite (A group) led to several unexpected effects such as an increase in clostridial population and serum alanine aminotrasferase and a decrease in haemoglobin concentration (P?<?0.05). The addition of alginite prevented a decrease in faecal pH and serum mineral content observed in the LF group. This indicates the possibility of applying alginite also in the nutrition of dogs as a combinative additive with probiotic bacteria for restoring optimal acid-alkali balance without affecting positive probiotic effects.     

Bacteriocin-producing strain of Enterococcus faecium EK 13 with probiotic character and its application in the digestive tract of rabbits

Enterococcus faecium EK 13 is a bacteriocin-enterocin A producing strain with probiotic properties. In this study its colonization, stability and effect on microflora in rabbits was studied as well as its influence on zootechnical parameters. Fifty rabbits of both sexes (HYPLUS, 30-day old; after weaning) were divided into control (CG) and experimental (EG) groups. They were fed a standard diet. Moreover, 25 rabbits in EG were fed daily (for 4 weeks) 15 g (separate doses ∼1.6 g) of lyophilized EK13 strain (rifampicin resistant variant — rif^R; 10⁹ cfu/g) dissolved in drinking water. After cessation of EK13 (rif^R) strain application, the rabbits in both groups were fed a standard diet for the next 2 weeks. Sampling was performed in double on day 0 (at the beginning of experiment), weekly during EK13 (rif^R) strain application as well as on week 1 and 2 after cessation of EK13 (rif^R) strain application. The counts of EK13 (rif^R) strain reached 7.1 ± 2.6 log₁₀ cfu/g after 4 weeks and even on week 2 after its cessation the counts 5.6 ± 2.3 log₁₀ cfu/g were determined. The total counts of enterococci in the rabbits were already increased in EG comparing with CG (p < 0.05); even 2 weeks after EK13 (rif^R) strain cessation, their counts in EG were 7.2 ± 2.6 log₁₀ cfu/g (p < 0.001). Enterococci in CG reached at the same time the value 3.7 ± 2.6 log₁₀ cfu/g. The counts of E. coli were significantly reduced in EG during 4 weeks (p < 0.05, p < 0.001). Even 2 weeks after EK13 (rif^R) strain cessation significant difference in E. coli counts between CG and EG was detected (p < 0.001). Enterobacteria in EG were significantly reduced (p < 0.001). Average daily gain in EG was 41.0 ± 3.83 in comparison to CG (40.6 ± 3.72); it means almost the same; although rabbits in EG showed higher feed intake per kg of gain than rabbits in CG. Preliminary results demonstrated that EK13 is a perspective probiotic candidate for rabbits. Presented at the Second Probiotic Conference, Košice, 15–19 September 2004, Slovakia.     

Biological control of crown rot of bananas with Pichia anomala strain K and Candida oleophila strain O

The antagonistic activity of two yeast strains (Pichia anomala (E.C. Hansen) Kurtzman, strain K and Candida oleophila Montrocher, strain O) against the parasitic complex responsible for banana crown rot was evaluated. The strains were applied at three different concentrations (10⁶, 10⁷, 10⁸ cfu/ml) and their efficacy tested in vivo on three separate fungi (Colletotrichum musae (Berk. & Curt.) Arx, Fusarium moniliforme Sheldon, and Cephalosporium sp.) and on a parasitic complex formed by association of these three fungi. At the concentrations used C. musae appeared to be the most pathogenic. The complex showed intermediate aggressiveness between C. musae and both other fungi.Statistically significant antagonistic effects were observed on C. musae, F. moniliforme, and the fungal complex. The highest protection level (54.4%) was observed with strain O added at 10⁸ cfu/ml on crowns previously inoculated with the fungal complex. The level was lower when the fungi were inoculated separately.Furthermore, the antagonistic effect was strongly reinforced when strain O at 10⁸ cfu/ml was applied 24 h before fungal complex inoculation (59.9%), as compared to its application 15 min (24.3%) or 3 h (27.3%) after fungal complex inoculation. Bananas showed increased susceptibility to the fungal complex from March to June, and this influenced the level of protection by yeast, which decreased over the same period. A strict negative correlation (R² = 0.83) was highlighted between susceptibility of banana to crown rot and protection provided by yeast.     

Lactobacillus fermentum ZYL0401 Attenuates Lipopolysaccharide-Induced Hepatic TNF-α Expression and Liver Injury via an IL-10- and PGE2-EP4-Dependent Mechanism

Lipopolysaccharide (LPS) has essential role in the pathogenesis of D-galactosamine-sensitized animal models and alcoholic liver diseases of humans, by stimulating release of pro-inflammatory mediators that cause hepatic damage and intestinal barrier impairment. Oral pretreatment of probiotics has been shown to attenuate LPS-induced hepatic injury, but it is unclear whether the effect is direct or due to improvement in the intestinal barrier. The present study tested the hypothesis that pretreatment with probiotics enables the liver to withstand directly LPS-induced hepatic injury and inflammation. In a mouse model of LPS-induced hepatic injury, the levels of hepatic tumor necrosis factor-alpha (TNF-α) and serum alanine aminotransferase (ALT) of mice with depleted intestinal commensal bacteria were not significantly different from that of the control models. Pre-feeding mice for 10 days with Lactobacillus fermentum ZYL0401 (LF41), significantly alleviated LPS-induced hepatic TNF-α expression and liver damage. After LF41 pretreatment, mice had dramatically more L.fermentum-specific DNA in the ileum, significantly higher levels of ileal cyclooxygenase (COX)-2 and interleukin 10 (IL-10) and hepatic prostaglandin E2 (PGE₂). However, hepatic COX-1, COX-2, and IL-10 protein levels were not changed after the pretreatment. There were also higher hepatic IL-10 protein levels after LPS challenge in LF41-pretreaed mice than in the control mice. Attenuation of hepatic TNF-α was mediated via the PGE₂/E prostanoid 4 (EP4) pathway, and serum ALT levels were attenuated in an IL-10-dependent manner. A COX-2 blockade abolished the increase in hepatic PGE₂ and IL-10 associated with LF41. In LF41-pretreated mice, a blockade of IL-10 caused COX-2-dependent promotion of hepatic PGE₂, without affecting hepatic COX-2levels. In LF41-pretreated mice, COX2 prevented enhancing TNF-α expression in both hepatic mononuclear cells and the ileum, and averted TNF-α-mediated increase in intestinal permeability. Together, we demonstrated that LF41 pre-feeding enabled the liver to alleviate LPS-induced hepatic TNF-α expression and injury via a PGE₂-EP4- and IL-10-dependent mechanism.     

A Study of Air Microbe Levels in Different Areas of a Hospital

Airborne transmission is an important route for many microbial pathogens in outdoor and indoor environments, including hospitals. A 2-year-long survey of bioaerosol quality in operating theatres (OT), hospital rooms (HR) and maternity wards (MW) at a hospital in Murcia, Spain, was performed. Total aerobic counts (TAC) and fungal load (FL) were assessed using a microbiological air sampler (MAS-100 single-stage impactor). While fungal levels were below 1 cfu/m³ (0–7.33 cfu/m³) in OT, they were higher in MW (mean, 6.9 cfu/m³; range 0.44–44.67 cfu/m³) and in HR (mean, 10.6 cfu/m³; range, 0–266 cfu/m³). In OT the aerobic counts were considerably higher, with a mean of 25.6 cfu/m³ (range, 1.67–157 cfu/m³). MW and HR also showed higher means for total aerobic counts compared to OT. Seasonal changes were not detected in mould and bacteria levels in OT. Hospital renovation occurred during this study and OT adjacent to renovated areas were closed. A survey of TAC and FL in OT resumed when renovation was completed. We observed an outstanding increase in FL (more than 100 cfu/m³), particularly Aspergillus spp., during this period, but no significant changes in TAC were observed after renovation.     

Positive legacy effect of previous legume proportion in a ley on the performance of a following crop of Lolium multiflorum

Aims

We investigated the legacy effects of a previous ley’s legume proportion on the performance of a following grass crop in a rotation.

Methods

In April 2015, a pure Lolium multiflorum L. crop was sown after the removal of legume containing swards (0–100% legumes), and was harvested four times over the following one-year period (3 times in 2015 and once the following April 2016). Labeled ¹⁵N fertilizer (50 kg N ha⁻¹) was applied during the 2nd and 3rd re-growth periods to determine N fluxes.

Results

Across the one-year period, a significant legume-legacy induced increase in biomass yield of L. multiflorum was observed over the entire range of previous legume proportions when compared against the non-legume ley, the effect being 2.15 and 1.73 t ha⁻¹ (P ≤ 0.001 each) in swards with 50% and 100% previous legume proportion, respectively, or up to +31%. The legume-legacy effect on biomass yield was most pronounced at the 1st harvest (June) and persisted into the 2nd harvest in August (P ≤ 0.05 both, over the entire range of previous legume proportion), though was no longer evident at the 3rd harvest (September). Importantly, the legume-legacy effect returned in the 4th harvest in April (P ≤ 0.05). Examining the source of N contributing to N yield confirmed that more N was derived from the soil at harvest 1 and 2 for previous legume containing leys (P ≤ 0.001) compared to those which contained no legumes, with a significant increase still seen for legume mixtures at harvest 3 (P ≤ 0.01).

Conclusions

The results demonstrate a sustained soil-transferred performance-enhancing legacy effect on a following crop in a rotation, with previous legume proportions of 50% having a comparable effect compared with that of a previous legume monoculture.

     

Lactobacillus fermentum BCS87 expresses mucus‐ and mucin‐binding proteins on the cell surface

Aims: To identify and characterize adhesion‐associated proteins in the potential probiotic Lactobacillus fermentum BCS87. Methods and Results: Protein suspensions obtained from the treatment of Lact. fermentum BCS87 with 1 mol 1^?1 LiCl were analysed by Western blotting using HRP‐labelled porcine mucus and mucin. Two adhesion‐associated proteins with relative molecular weight of 29 and 32 kDa were identified. The N‐terminal and internal peptides of the 32 kDa protein (32‐Mmubp) were sequenced, and the corresponding gene (32‐mmub) was found by inverse polymerase chain reaction. The complete nucleotide sequence of 32‐mmub revealed an open reading frame of 903 bp encoding a primary protein of 300 amino acids and a mature protein of 272 residues. A basic local alignment search showed 47–99% identity to solute‐binding components of ATP binding cassette transporter proteins in Lactobacillus, Streptococcus and Clostridium. An OpuAC‐conserved domain was identified and phylogenetic relationship analysis confirmed that 32‐Mmubp belongs to the OpuAC family. Conclusions: Adhesion of Lact. fermentum BCS87 appeared to be mediated by two surface‐associated proteins. 32‐Mmubp is a component of ABC transporter system that also functions as an adhesin. Significance and Impact of the Study: Characterization of 32‐Mmubp and 32‐mmub will contribute to understanding the host–bacteria interactions of Lact. fermentum with the intestinal tract of pigs.     

Evaluation of bio-aerosols at an animal feed manufacturing industry: A case study

Both total and biological particles (totalculturable bacteria, Gram negative bacteria,mold and actinomycetes) were measured at ananimal feed manufacturing industry. Suspendedparticle concentration ranged from 1.72 to2.3 mg m^–3 with a mean value of1.97 mg m^–3. Airborne microorganisms weredetected in lower concentrations than thoseassociated with suspended dust andfeed-materials. Bacterial concentrations weretwo to three times higher than concentrationsof mold and actinomycetes. Bacterialconcentrations averaged4.86 × 10³ cfu m^–3; 2.6 × 10⁴cfu m^–3 and 3.96 × 10⁷ cfu g^–1 inair, associated with suspended dust andfeed-materials, respectively, whereas moldconcentrations averaged 7.33 × 10²cfu m^–3; 1.97 × 10³ cfu m^–3 and7 × 10⁵ cfu g^–1 of the correspondingenvironments, respectively. Enterobacterspp and Klebsiella spp were the mostabundant Gram negative bacteria, whereas Bacillus species. were the most dominant Grampositive bacteria. Aspergllius niger,other Aspergillus species and Penicillium were the dominant mold isolates.Acremonium was only detected in feedmaterials, whereas Aspergillus fumigatuswas only detected in air. The animal feedindustry environment has a significantbio-contamination and many of microorganismsimplicated in respiratory problems weredetected in this environment.     

Biofunctional Attributes of Surface Layer Protein and Cell-Bound Exopolysaccharide from Probiotic Limosilactobacillus fermentum (MTCC 5898)

The probiotic extracellular matrix components (ECM) have been considered as an important factor in eliciting the beneficial roles of the bacteria. The study involved the growth phase–dependent extraction of the surface layer protein (SLP) and cell-bound exopolysaccharide (EPS-b) from novel Limosilactobacillus fermentum (MTCC 5898). Both SLP and EPS-b were optimally extracted at the late logarithmic phase of the bacteria upon 8 h of incubation. Furthermore, the adhesive, immunomodulatory, and anti-oxidative potential of the extracted components were evaluated using in vitro models. The major role of SLP was evidenced on bacterial adhesion to mucin and was related to its hydrophobic character. Under in vitro conditions, no effect of SLP and EPS-b was observed on the proliferation of murine splenocytes; however, both the components stimulated the phagocytosis of murine peritoneal macrophages at varying concentrations. Furthermore, all the components exhibited strong radical scavenging, chelating, and reducing potential with increasing concentration. Therefore, the ECM components of L. fermentum exhibited a variable biofunctional effect, providing crucial information to enable its further use as functional foods and overcome the challenges posed by probiotics.