Oral Administration of Lactobacillus Strains Isolated from Breast Milk as an Alternative for the Treatment of Infectious Mastitis during Lactation

In this study, 20 women with staphylococcal mastitis were randomly divided in two groups. Those in the probiotic group daily ingested 10 log₁₀ CFU of Lactobacillus salivarius CECT5713 and the same quantity of Lactobacillus gasseri CECT5714 for 4 weeks, while those in the control one only ingested the excipient. Both lactobacillus strains were originally isolated from breast milk. On day 0, the mean staphylococcal counts in the probiotic and control groups were similar (4.74 and 4.81 log₁₀ CFU/ml, respectively), but lactobacilli could not be detected. On day 30, the mean staphylococcal count in the probiotic group (2.96 log₁₀ CFU/ml) was lower than that of the control group (4.79 log₁₀ CFU/ml). L. salivarius CECT5713 and L. gasseri CECT5714 were isolated from the milk samples of 6 of the 10 women of the probiotic group. At day 14, no clinical signs of mastitis were observed in the women assigned to the probiotic group, but mastitis persisted throughout the study period in the control group women. In conclusion, L. salivarius CECT5713 and L. gasseri CECT5714 appear to be an efficient alternative for the treatment of lactational infectious mastitis during lactation.     

Effect of probiotic L. plantarum IS-10506 and zinc supplementation on humoral immune response and zinc status of Indonesian pre-school children

A 90-day randomized, double-blind, placebo-controlled, pre-post trial was conducted in four groups of Indonesian children aged 12–24 months: placebo, probiotic, zinc, and a combination of probiotic and zinc (n = 12 per group). Microencapsulated Lactobacillus plantarum IS-10506 of dadih origin was supplemented at a dose of 10¹⁰ CFU/day as a probiotic. Zinc was supplemented as 20 mg zinc sulfate monohydrate (8 mg zinc elemental). Blood and stool samples were collected at baseline and at the end of the study period. Fecal sIgA was assessed by ELISA and serum zinc concentrations by ICP-MS. Fecal sIgA increased significantly in the probiotic group (30.33 ± 3.32 μg/g; p < 0.01) and in the combination probiotic and zinc group (27.55 ± 2.28 μg/g; p < 0.027), as compared with the placebo group (13.58 ± 2.26 μg/g). Changes in serum zinc concentrations in the combination probiotic and zinc group showed the highest elevation at the end of the study period. A combination of probiotic L. plantarum IS-10506 at a dose of 10¹⁰ CFU/day and 8 mg of elemental zinc supplementation showed a potential ability to improve the zinc status of pre-school children. Taken together, supplementation with the probiotic L. plantarum IS-10506 and zinc for 90 days resulted in a significantly increased humoral immune response, as well as improved zinc status, in young children.     

Influences of short-term pre-slaughter dietary manipulation in sheep and goats on pH and microbial loads of gastrointestinal tract

Sheep (BW = 39.9 kg, n = 16) and goats (BW = 32.8 kg, n = 16) were used in a completely randomized design to determine the effect of short-term pre-slaughter diet and feed deprivation (FD) time on pH and microbial loads in the gastrointestinal tract (GIT) contents. In a 2 × 2 × 2 factorial treatment arrangement, the main effects of species, diet, and FD time prior to slaughter and their interactions were studied. Animals were fed either a hay or concentrate diet for 4 d and then feed deprived for either 12 or 24-h prior to slaughter. The pH of rumen and colon contents as well as weight of GIT was measured. The contents of rumen and rectum were also sampled for microbial analysis. The GIT of sheep (1.82 kg) was heavier (P < 0.05) than that of goats (1.46 kg). The 12-h FD group (1.74 kg) had a higher (P < 0.05) GIT weight than the 24-h FD group (1.53 kg). Hay-fed animals had higher (P < 0.05) rumen (7.08 vs. 6.43) and colon pH values (7.02 vs. 6.56) than those of the concentrate-fed animals. The 24-h FD group (3.39 ± 0.272 log₁₀CFU/g) contained more (P < 0.05) Escherichia coli in the rumen than did the 12-h FD (2.17 ± 0.272 log₁₀CFU/g) group. The concentrate-fed animals (3.49 ± 0.289 log₁₀CFU/g) had higher (P < 0.05) coliform counts in the rumen than the hay-fed animals (2.43 ± 0.289 log₁₀CFU/g). The 24-h FD group (3.42 ± 0.289 log₁₀CFU/g) had a higher (P < 0.05) concentration of coliform than did the 12-h FD group (2.50 ± 0.289 log₁₀CFU/g). The 24-h FD group (3.31 ± 0.259 log₁₀CFU/g) also had higher (P < 0.05) Enterobacteriaceae counts in the rumen than did in the 12-h FD group (2.47 ± 0.259 log₁₀CFU/g). Goats (5.71 ± 0.158 log₁₀CFU/g) had lower (P < 0.05) total plate counts in the rumen compared to sheep (6.27 ± 0.158 log₁₀CFU/g). The concentrate-fed animals had higher (P < 0.05) E. coli (6.44 vs. 4.01 ± 0.468 log₁₀CFU/g), total coliform (6.74 vs. 4.16 ± 0.469 log₁₀CFU/g), Enterobacteriaceae (6.93 vs. 3.83 ± 0.651 log₁₀CFU/g), and total plate counts (7.79 vs. 7.28 ± 0.170 log₁₀CFU/g) in the rectum than the hay-fed animals. The results indicate that microbial loads in the GIT of small ruminants may be reduced by either feeding hay for 4 d or depriving feed for 12-h prior to slaughter.     

Lowering of oxidative stress improves endothelial function in healthy subjects with habitually low intake of fruit and vegetables: A randomized controlled trial of antioxidant- and polyphenol-rich blackcurrant juice

Inadequate intake of the recommended five-a-day fruit and vegetable portions might contribute to increased cardiovascular disease risk. We assessed the effects of dietary intake of a blackcurrant juice drink, rich in vitamin C and polyphenols, on oxidative stress and vascular function. This was a double-blind, placebo-controlled, parallel group study of 66 healthy adults who habitually consume <2 portions of fruit and vegetables per day. Participants were randomly allocated to consume 250 ml of placebo (flavored water) or low or high blackcurrant juice drink four times a day for 6 weeks. Flow-mediated dilation (FMD) and plasma concentrations of F₂-isoprostanes and vitamin C were measured. In the high blackcurrant juice drink group FMD increased significantly (5.8±3.1 to 6.9±3.1%, P=0.022) compared with the placebo group (6.0±2.2 to 5.1±2.4%). Plasma vitamin C concentration increased significantly in the low (38.6±17.6 to 49.4±21.0 µmol/L, P<0.001) and high (34.6±20.4 to 73.8±23.3 µmol/L, P<0.001) blackcurrant juice drink groups compared with the placebo group (38.1±21.0 to 29.0±17.6 µmol/L). F₂-isoprostane concentrations were significantly lower in the high blackcurrant juice drink group (225±64 pg/ml) compared with the low blackcurrant juice drink (257±69 pg/ml, P=0.002) and placebo group (254±59 pg/ml, P=0.003). At follow-up, changes in plasma vitamin C correlated significantly with changes in FMD (r=0.308, P=0.044). Consumption of blackcurrant juice drink high in vitamin C and polyphenols can decrease oxidative stress and improve vascular health in individuals with habitually low dietary fruit and vegetable intake.     

Complete Genome Sequence of Lactobacillus salivarius CECT 5713, a Probiotic Strain Isolated from Human Milk and Infant Feces

Lactobacillus salivarius is a homofermentative lactic acid bacterium and is frequently isolated from mucosal surfaces of healthy humans. L. salivarius CECT 5713, a strain isolated simultaneously from breast milk and infant feces of a healthy mother-infant pair, has immunomodulatory, anti-inflammatory, and anti-infectious properties, as revealed by several in vitro and in vivo assays. Here, we report its complete and annotated genome sequence.In the last years, culture-dependent and -independent analyses of the bacterial diversity of human milk and colostrum have revealed that these biological fluids are a source of live staphylococci, streptococci, lactic acid bacteria, and bifidobacteria in the infant gut (5, 6, 8, 9, 11, 13), where they play a key role in the initiation and development of the gut microbiota (12). In a previous study, we isolated L. salivarius CECT 5713 from human milk and infant feces of a mother-child pair (10). Subsequent studies revealed that this strain was a good probiotic candidate since it achieved high survival rates when exposed to the gastrointestinal tract conditions, showed a strong adherence to intestinal cells, stimulated the expression of mucin-encoding genes, produced antimicrobial compounds (lactate, acetate, and hydrogen peroxide), and displayed in vivo and in vitro immunomodulatory, anti-inflammatory, and antibacterial properties against pathogenic bacteria (2, 10, 15). Moreover, oral administration of L. salivarius CECT 5713 appears to be an efficient alternative for the treatment of infectious mastitis in lactating women (7). Similarly, studies with other L. salivarius strains in animal models and clinical trials have demonstrated their probiotic function and, particularly, their anti-inflammatory effects (3, 14, 16).In order to interrogate the genome sequence of L. salivarius CECT 5713 with regard to its probiotic properties, the complete genome sequence was determined by a whole-genome shotgun strategy using pyrosequencing technology (454 Life Sciences, Banford, CT). The initial draft assembly provided by 454 Life Sciences was based on 444,604 high-quality pyrosequencing reads, which assembled into 59 contigs. The genome sequence of L. salivarius UCC118 (1), a well-characterized probiotic strain, was used to order these contigs into large scaffolds.The genome of L. salivarius CECT 5713 consists of a circular chromosome of 1,828,169 bp, two plasmids (pHN1, 44,581 bp; pHN2, 20,426 bp), and a megaplasmid (pHN3, 242,962 bp). The overall GC content of the chromosome is 32.93%, similar to that of the megaplasmid but lower than those of the plasmids (>38%). The entire genome of CECT 5713 contains 1,558 protein-, 87 tRNA-, and 51 rRNA-encoding genes. A comparison between the genomes of L. salivarius CECT 5713 and UCC118 revealed the presence of 52 protein-encoding genes that are exclusive for CECT 5713, including genes encoding a 6-phospho-β-glucosidase and three collagen-binding proteins, which may explain the high potential for competitive exclusion of pathogens displayed by this strain. The genes responsible for the bacteriocin activity of L. salivarius CECT 5713 are located in pHN3. This megaplasmid contains six open reading frames (ORFs) closely related, but not identical, to the genes responsible for the biosynthesis of salivaricin ABP-118, a two-component class II bacteriocin (4), in L. salivarius UCC118. Globally, several features of the L. salivarius CECT 5713 genome suggest a strong probiotic potential in humans.     

Reference database of the gait cycle for young healthy Tunisian adults

BackgroundQuantified gait analysis is a rising technology used increasingly to assess motor disorders. Normal reference data are required in order to evaluate patients, but there are no reference data available for the Tunisian healthy population.AimTo assess the features of normal Tunisian gait pattern, and examine the intrinsic reliability of spatio-temporal, kinematic and kinetic parameters within a new specific reference database.MethodsEighteen healthy active-young adults (age: 23.30 ± 2.54 years, height: 1.78 ± 0.04 m and, weight: 70.00 ± 4.80 kg) have participated to five trials of step gait where the dominant lower limb were recorded. Two over the five trials were randomly selected to be further analyzed. Twenty-three spatio-temporal, kinematic and kinetic parameters determined from 3-dimensional gait analysis. The intrinsic reliability was examined for each variable and our results were compared with those available in the literature.ResultsTwelve over 23 parameters have an excellent intrinsic reliability (P > 0.05, ICC > 0.9 and SEM < 5% of the grand mean). There are similarities with other studies (P < 0.05) but we noticed the existence of some specificity (the height of hip extension peak and the low cadence of gait) that could characterize the Tunisian population.ConclusionA specific reference database of the gait cycle has been established for healthy Tunisian active-young adults and excellent inter-trial reliability may be observed for different variables.     

Enhancing bio-suppression of Parthenium hysterophorus L.: Diapause in Zygogramma bicolorata Pallister and its manipulation through insulin-like peptides (ILPs)

In-depth investigations on diapause behaviour of Z. bicolorata revealed that the adults entered diapause at any time from August to December and that the number peaked (42.00%) during the last half of November. The percentage of adults entering diapause increased with a decrease in day length. Weight of diapausing adults was significantly higher than weight of non-diapausing adults. The percentage of adults undergoing diapause at 30 °C was significantly lower than those undergoing diapause at 15 and 25 °C. The percentage of adults burrowing increased with increasing moisture. In silty soil and soil with high organic matter, 46.7% and 49.2% of adults entered diapause, respectively, whereas in sandy soil, only 23.5% of adults entered diapause. When newly emerged beetles were exposed to 5 μg of human insulin 30/70, a significantly lower percentage of treated adults underwent diapause compared to untreated adults under both feeding and no feeding conditions. Insulin treatment also influenced the emergence period from diapause (93.92 ± 1.73 days), percent emergence (81 ± 1.54%) and fecundity/month (512.7 ± 25.38 eggs) of Z. bicolorata in treated adults as compared to untreated adults (109.05 ± 2.2, 74.00 ± 1.82 and 438.3 ± 19.33 eggs, respectively). However, there was no significant impact of insulin on adult longevity. These findings are of great utility in the biological suppression of Parthenium as it will enhance the effectiveness of this beetle through manipulation of diapause.     

Menstrual Disorders and Androgen-Related Traits in Young Women with Type 1 Diabetes Mellitus: a Clinical Study

Objective: To investigate possible causes of menstrual disorders and androgen-related traits in young women with type 1 diabetes mellitus (T1DM).Methods: Fifty-three women with T1DM (duration 8.0 ± 5.6 years), 41 women with (polycystic ovary syndrome) PCOS, and 51 controls matched for age (19.4 ± 4.3 years vs. 21.2 ± 2.7 years vs. 20.8 ± 3.1 years; P>.05) and body mass index (BMI) (22.2 ± 2.7 kg/m² vs. 21.9 ± 2.0 kg/m² vs. 21.4 ± 1.9 kg/m²; P>.05) were prospectively recruited.Results: Two women (3.8%) in the T1DM group had not experienced menarche (at 15.5 and 16.6 years); of the rest, 23.5% had oligomenorrhea, 32.1% hirsutism, and 45.3% had acne. The age at menarche was delayed in the T1DM group compared to controls (12.7 ± 1.3 vs. 12.0 ± 1.0 years; P = .004), while no difference was observed with the polycystic ovary syndrome (PCOS) group (12.4 ± 1.2 years). There were no differences in total testosterone (0.43 ± 0.14 ng/mL vs. 0.39 ± 0.14 ng/mL; P>.05), dehydroepiandrosterone sulfate (DHEA-S) (269 ± 112 μg/dL vs. 238 ± 106 μg/dL; P>.05) or Δ4-androstenedione (2.4 ± 1.3 ng/mL vs. 1.9 ± 0.5 ng/mL; P>.05) concentrations between T1DM and controls. However, patients with T1DM had lower sex hormone binding globulin (SHBG) concentrations than controls (61 ± 17 nmol/L vs. 83 ± 18.1 nmol/L; P = .001), which were even lower in the PCOS group (39.5 ± 12.9 nmol/L; P = .001 compared with T1DM). The free androgen index (FAI) was higher in the PCOS group compared with both other groups (T1DM vs. PCOS vs. controls: 2.53 ± 0.54 vs. 7.88 ± 1.21 vs. 1.6 ± 0.68; P<.001). FAI was higher in patients with T1DM compared to controls as well (P = .038). There was no difference in DHEA-S concentrations between T1DM and PCOS patients (269 ± 112 μg/dL vs. 297 ± 100 μg/dL; P>.05).Conclusion: Menstrual disorders and androgen-related traits in young women with T1DM may be attributed to an increase in androgen bioavailability due to decreased SHBG concentrations.     

Lower dipeptidyl peptidase-4 following exercise training plus weight loss is related to increased insulin sensitivity in adults with metabolic syndrome

Dipeptidyl peptidase-4 (DPP-4) is a circulating glycoprotein that impairs insulin-stimulated glucose uptake and is linked to obesity and metabolic syndrome. However, the effect of exercise on plasma DPP-4 in adults with metabolic syndrome is unknown. Therefore, we determined the effect of exercise on DPP-4 and its role in explaining exercise-induced improvements in insulin sensitivity. Fourteen obese adults (67.9 ± 1.2 years, BMI: 34.2 ± 1.1 kg/m²) with metabolic syndrome (ATP III criteria) underwent a 12-week supervised exercise intervention (60 min/day for 5 days/week at ∼85% HR_max). Plasma DPP-4 was analyzed using an enzyme-linked immunosorbent assay. Insulin sensitivity was measured using the euglycemic-hyperinsulinemic clamp (40 mU/m²/min) and estimated by HOMA-IR. Visceral fat (computerized tomography), 2-h glucose levels (75 g oral glucose tolerance), and basal fat oxidation as well as aerobic fitness (indirect calorimetry) were also determined before and after exercise. The intervention reduced visceral fat, lowered blood pressure, glucose and lipids, and increased aerobic fitness (P < 0.05). Exercise improved clamp-derived insulin sensitivity by 75% (P < 0.001) and decreased HOMA-IR by 15% (P < 0.05). Training decreased plasma DPP-4 by 10% (421.8 ± 30.1 vs. 378.3 ± 32.5 ng/ml; P < 0.04), and the decrease in DPP-4 was associated with clamp-derived insulin sensitivity (r = −0.59; P < 0.04), HOMA-IR (r = 0.59; P < 0.04) and fat oxidation (r = −0.54; P < 0.05). Increased fat oxidation also correlated with lower 2-h glucose levels (r = −0.64; P < 0.02). Exercise training reduces plasma DPP-4, which may be linked to elevated insulin sensitivity and fat oxidation. Maintaining low plasma DPP-4 concentrations is a potential mechanism whereby exercise plus weight loss prevents/delays the onset of type 2 diabetes in adults with metabolic syndrome.     

Bifidobacteria in the digestive tract of bumblebees

Bifidobacteria and other bacterial groups (lactobacilli, facultative anaerobes, anaerobes) from the digestive tract of three bumblebee species (Bombus lucorum (34 samples), Bombus pascuorum (18 samples) and Bombus lapidarius (9 samples)) were enumerated and characterised. Counts of facultative anaerobic bacteria and lactobacilli (5.41 ± 2.92 and 2.69 ± 3.02 log CFU/g of digestive tract content) were lower than those of anaerobes (7.66 ± 0.86 log CFU/g). Counts of bifidobacteria were determined using two selective media: MTPY (Modified Trypticase Phytone Yeast extract agar) and a new medium with pollen extract. There was no significant difference between the counts of bifidobacteria from both media, 5.00 ± 2.92 log CFU/g on MTPY and 5.00 ± 2.87 on the pollen medium. Subsequently, 187 bacterial strains of the family Bifidobacteriaceae (fructose-6-phosphate phosphoketolase-positive) were isolated from three different localities and from all three species of bumblebees. Bifidobacteria were found in 42 out of 61 specimens (69%). Twenty-three (38%) specimens had counts of bifidobacteria higher than 7.0 log CFU/g. Bifidobacteria represented the dominant group of anaerobes (>70% of total anaerobes), i.e., the principal group of bacteria in the bumblebee digestive tract, in only fourteen specimens (23% of total). For the first time, bifidobacteria were isolated from the digestive tract of bumblebees. In addition, we suggest, on the basis of biochemical tests (API 50 CHL and RAPID ID 32) and genetic methods (PCR and DGGE), that these bacteria may represent new species within the family of Bifidobacteriaceae.     

Dogs with Leishmania chagasi infection have semen abnormalities that partially revert during 150 days of Allopurinol and Amphotericin B therapy

The goal of the present study was to characterize the semen quality of dogs naturally infected with Leishmania chagasi, and treated with Allopurinol and Amphotericin B. Eight naturally infected and eight non-infected dogs were selected. Following semen collection, progressive motility, vigor, concentration and sperm morphology were evaluated. The seminal patterns in the treated animals were evaluated at the beginning (d0) and at days 30 (d30), 60 (d60) and 150 (d150) of treatment. The progressive motility at d0 (35.7 ± 22.3%) was less than that of the control group (77.8 ± 7.1%) (P < 0.05). The vigor was similar to the control group throughout the treatment (P > 0.05). The number of sperm/mL, sperm/ejaculate and sperm/kg of body weight was similar among groups (P > 0.05). The percentages of normal spermatozoa of infected and treated animals were similar throughout the treatment and to the control group (69.1 ± 8.7%) at d60 (37.5 ± 11.2%) and d150 (48.3 ± 10.8%) (P > 0.05), but smaller at d0 (22.7 ± 10.5%) and d30 (28.8 ± 15.9%) (P < 0.05). A greater percentage of acrosome damage was observed in the control group (3.1 ± 2.3%) compared to the d60 (0.1 ± 0.2%) (P < 0.05). The infected dogs had a greater percentage of principal piece defects at d60 (37.0 ± 6.3%) than the control group (16.8 ± 7.3%) (P < 0.05); and greater percentages of detached normal heads at d0 (28.7 ± 19.7%) and d30 (18.5 ± 18.5%) than the control group (0.4 ± 0.5%) (P < 0.05). This reduction in semen quality of the infected animals is suggestive of an epididymal dysfunction. Due to this poor semen quality, caution is recommended when using infected male dogs for reproductive purposes.     

Biochemical changes in the follicular fluid of the dominant follicle of high producing dairy cows exposed to heat stress early post-partum

High yielding dairy cows experience a negative energy balance (NEB) early post-partum and it was hypothesized that this may be aggravated under summer heat stress (HS) conditions. In this study, which was performed in Egypt, 20 Holstein cows were followed during summer (n = 10) and winter (n = 10) seasons. All cows were multiparous and kept at the same herd. Blood was sampled from each cow starting 1 week before the expected calving date and then at 1-week intervals until week 6 post-partum. From week 2 to 6 post-partum follicular fluid was collected through transvaginal follicular fluid aspiration at 6 days intervals. Ambient air temperature (AT) and relative humidity (RH) were recorded and temperature–humidity index (THI) was calculated as well. Respiration rate (RR), rectal temperature (RT), and body condition score (BCS) were recorded for each cow at the time of blood sampling. Concentrations of glucose, insulin like growth factor-1 (IGF-1), non-esterified fatty acids (NEFA), urea and total cholesterol (TC) were measured in each blood and follicular fluid sample. All the cows showed a significantly higher RR and RT in summer (95.5 ± 1.1 and 39.88 ± 0.06, respectively) than in winter (43.89 ± 0.61 and 38.94 ± 0.07, respectively) (P < 0.001). Body condition score loss during the early post-partum period was higher in summer than in winter (1.1 ± 0.07 vs. 0.85 ± 0.06 point, respectively) (P < 0.001). The average dominant follicle diameter was significantly lower in summer than in winter during the period of negative energy balance (11.6 ± 0.7 mm vs. 15.3 ± 1.2 mm, respectively) (P < 0.01). Under summer heat stress, the concentrations of glucose (2.98 ± 0.07 and 2.19 ± 0.04 mmol/L), IGF-1 (106.7 ± 2.9 and 99.0 ± 3.4 ng/ml) and TC (137.3 ± 5.3 and 62.2 ± 5.1 mg/dl) in blood and FF, respectively, were significantly lower than winter concentrations by (0.17 ± 0.03 mmol/L, P < 0.001 and 0.26 ± 0.06 mmol/L, P < 0.001), (12.3 ± 3.6 ng/ml, P < 0.001 and 9.0 ± 2.7 ng/ml, P < 0.001) and (20.7 ± 1.8 mg/dl, P < 0.001 and 7.3 ± 1.1 mg/dl, P < 0.01), respectively. However, the concentrations of NEFA (0.68 ± 0.14 and 0.22 ± 0.02 mmol/L) and urea (9.27 ± 0.34 and 9.96 ± 0.25 mmol/L) in blood and FF, respectively, were significantly higher in summer compared to winter (0.50 ± 0.08 mmol/L, P < 0.001 and 0.20 ± 0.02 mmol/L, P < 0.001) and (8.77 ± 0.23 mmol/L, P < 0.05 and 8.96 ± 0.29 mmol/L, P < 0.001), respectively, throughout the experimental period. The results of the present study indicate that heat stress early post-partum aggravates NEB in high yielding dairy cows, reduces BCS, dominant follicle diameter and alters the biochemical concentrations in the follicular fluid of the dominant follicle which may result in inferior oocyte and granulosa cell quality and hence poorer fertility.     

A novel medium for the production of cephamycin C by Nocardia lactamdurans using solid-state fermentation

In this study, Nocardia lactamdurans NRRL 3802 was explored for the first time for production of cephamycin C by using solid-state fermentation. The effects of various substrates, moisture content, inoculum size, initial pH of culture medium, additional nitrogen source and amino acids were investigated for the maximum production of cephamycin C by N. lactamdurans NRRL 3802 in solid-state fermentation. Subsequently, selected fermentation parameters were further optimized by response surface methodology (RSM). The soybean flour as a substrate with moisture content of 65%, initial pH of culture medium of 6.5 and inoculum size of 10⁹ CFU/ml (2 × 10⁸ CFU/gds) at 28 ± 2 °C after 4 days gave maximum production of 15.75 ± 0.27 mg/gds of cephamycin C as compared to 8.37 ± 0.23 mg/gds before optimization. Effect of 1,3-diaminopropane on cephamycin C production was further studied, which further increased the yield to 27.64 ± 0.33 mg/gds.     

Effect of cryopreservation and microencapsulation of lactic acid bacterium Enterococcus faecium MC13 for long-term storage

The aim of this work was to investigate the effect of cryoprotectants on the survival of probiotic bacterium Enterococcus faecium MC13 during freeze drying and storage. The maximum relative cell viabilities were observed when cells were freeze dried and stored at −20 °C, which is optimum temperature for the preservation of E. faecium. At all storage temperatures, trehalose was found to be retaining the highest relative cell viability than other cryoprotectants. In addition, alginate–chitosan capsules were produced to encapsulate E. faecium with the aim of enhancing survival of probiotic cells and keeping the probiotic during exposure to the harsh gastro-intestinal conditions. Encapsulation of probiotic into alginate–chitosan capsules found to be retaining higher survival of probiotic cells (4.342 ± 0.26 Log CFU mL⁻¹) at −20 °C for six months. Microencapsulated cells were resistant to simulated gastric (pH 2.0) and intestinal fluids (pH 7.5), resulting in significantly enhanced survival when compared with free cells. During in vivo treatment, capsules were broken and probiotic cells were directly released into the intestinal tract of rat. This result showed that microencapsulation of E. faecium MC13 with alginate and a chitosan coating offers an effective means of delivery of viable cells to the colon and maintains their survival during the adverse gastro-intestinal conditions.     

The influence of pre-operative risk on the number of circulating endothelial progenitor cells during cardiopulmonary bypass

Background aimsThe number of circulating endothelial progenitor cells (EPC) depends on cytokine release and is also associated with cardiovascular risk factors. During cardiopulmonary bypass (CPB) the endothelium is the first organ to be affected by mechanical and immunologic stimuli. We hypothesized that the magnitude of EPC mobilization by CPB correlates with the pre-operative cardiovascular morbidity profile.MethodsEPC were quantified in blood samples from 30 patients who underwent cardiac surgery by magnetic bead isolation and fluorescence-activated cell sorting (FACS) analysis, based on concomitant expression of CD34, CD133 and CD309. Patients were divided into two groups based on the European System for Cardiac Operative Risk Evaluation (EuroSCORE): low risk (LR) and high risk (HR). Ten healthy volunteers served as controls. Samples were obtained before the start of CPB and at 1 and 24 h post-operatively. Plasma samples were collected for determination of release levels of cytokines and growth factors.ResultsAll CPB patients showed a significantly reduced basal number of EPC compared with healthy individuals (LR 5.60 ± 0.39/mL, HR 3.89 ± 0.34/ mL, versus control 0.807 ± 0.82/mL, P = 0.012 versus LR, P < 0.001 versus HR). CPB induced EPC release that peaked 1 h after surgery (pre-operative 4.79 ± 0.32/mL, 1 h 57.49 ± 5.31/mL, 24 h 6.67 ± 1.05/mL, P < 0.001 pre-operative versus 1 h, P < 0.001 pre-operative versus 24 h) and was associated with the duration of CPB. However, EPC release was significantly attenuated in HR patients (33.09 ± 3.58/mL versus 81.89 ± 4.36/mL at 1 h after CPB, P < 0.0001) and inversely correlated with the pre-operative EuroSCORE. Serum granulocyte–colony-stimulating factor (G-CSF), stem cell factor (SCF) and vascular endothelial growth factor (VEGF) levels increased throughout the observation period and were also correlated with the EPC count.ConclusionsCardiovascular risk factors influence the mobilization of EPC from the bone marrow after stimulation by CPB. This could be secondary to impaired mobilization or the result of increased EPC turnover, and may have implications for future cell therapy strategies in cardiac surgical patients.     

The effect of time left alone at home on dog welfare

The aim of this study was to investigate the effect of time left alone on dog behaviour and cardiac activity. Twelve privately owned dogs, with no history of separation related behaviour problems, were video-recorded on three different occasions when left alone in their home environment. The treatments lasted for 0.5 h (T_0.5); 2 h (T₂) and 4 h (T₄). Video-recording started 10 min before the owner left the house and continued until 10 min after the owner returned, so that interactions between dog and owner as well as behaviour during separation could be studied. Data on heart rate (HR) and heart rate variability (HRV) were collected within the same time period in each treatment. In addition to analysing behaviours separately, behaviours were also grouped together and defined as new variables; physically active, attentive behaviour, vocal, interaction initiated by owner and interaction initiated by dog. There were no differences in behaviour between treatments at equivalent time intervals until the owner returned, although a number of differences were observed at reunion with the owner. Dogs showed a higher frequency of physical activity (P < 0.05) and attentive behaviour (P < 0.01) in T₂ (0.37 ± 0.07; 0.52 ± 0.08, mean frequency of occurrence/15 s ± SE) and T₄ (0.48 ± 0.08; 0.48 ± 0.07) compared to T_0.5 (0.20 ± 0.07; 0.21 ± 0.05). They also showed more tail wagging (P < 0.01) and interacted more with their owners (P < 0.01) in T₂ (0.27 ± 0.08; 0.47 ± 0.09) and T₄ (0.26 ± 0.04; 0.42 ± 0.09) compared to T_0.5 (0.09 ± 0.04; 0.14 ± 0.03). After a longer time of separation, the dogs also showed higher frequencies of lip licking (P < 0.05) and body shaking (P < 0.05) at the owner's return (T_0.5 = 0.09 ± 0.05; T₂ = 0.24 ± 0.08; T₄ = 0.27 ± 0.06 and T_0.5 = 0.03 ± 0.01; T₂ = 0.08 ± 0.03; T₄ = 0.07 ± 0.01, respectively). There was a tendency for higher HR (P < 0.1) during the first and second minute after reunion in T₂ (127.6 ± 1.25, mean bpm ± SE; 111.3 ± 1.24) compared to T_0.5 (106.2 ± 1.06; 87.5 ± 1.02). According to the results of this study, the effect of time left alone was shown by a more intense greeting behaviour by the dog towards their owner as well as by a higher frequency of physical activity and attentive behaviour when the owner returned, already after 2 h of separation. Although this study cannot distinguish between whether dogs were aware of the length of time they were alone (but did not signal it) or whether they were unaware until reminded of it by the return of their owner, it does confirm that dogs are affected by the duration of time at home alone.     

Intra- and interday reliability of voluntary and electrically stimulated isometric contractions of the quadriceps femoris

The reliability of voluntary and electrically stimulated isometric contractions of m. quadriceps femoris of male participants (n = 10; age 30 ± 8 years; height 1.79 ± 0.05 m; body mass 79.4 ± 8.3 kg) was investigated using ratio limits of agreement (LoA) on a time scale common to examine recovery from muscle damaging exercise. No systematic changes in reliability occurred over time (baseline versus 2, 24, 48, and 72 h). Maximal voluntary contraction (MVC) and interpolated twitch technique (ITT) showed no mean bias (P > 0.05) with 95% LoA of ±12.7 and ±5.4, respectively. Resting twitch and potentiated doublet peak force showed no mean bias (P > 0.05). However, 95% LoA were smaller for the doublet (±13.9) than the twitch (±32.0). Twitch and doublet rates showed similar trends. Ratio of low (20 Hz) to high (50 Hz) frequency forces showed no mean bias (P > 0.05) and 95% LoA of (±9.2). However, there was significant mean bias (P < 0.05) and wider 95% LoA for peak force, contraction and relaxation parameters of the low and high frequency forces. In conclusion, MVC, ITT, potentiated doublet and the ratio of low to high frequency forces are recommended to most reliably examine functional muscle recovery between 2 and 72 h after damaging exercise.     

Relationship between plasma copeptin levels and complications of community-acquired pneumonia in preschool children

High plasma copeptin level has been associated with clinical outcomes after acute illness. The present study was undertaken to investigate the plasma copeptin concentrations in preschool children with community-acquired pneumonia (CAP) and to analyze the correlations of copeptin with CAP-related complications and pleural effusion. Plasma copeptin concentrations of 100 healthy children and 165 preschool children with CAP were measured. 35 children (21.2%) presented with complicated CAP and 28 children (17.0%) presented with pleural effusion. The admission copeptin levels were significantly increased in all patients (49.7 ± 21.4 pmol/L), children with complicated CAP (73.0 ± 16.9 pmol/L), those with uncomplicated CAP (43.4 ± 17.8 pmol/L), those with pleural effusion (70.9 ± 17.4 pmol/L) and those without pleural effusion (45.3 ± 19.5 pmol/L) compared with healthy control individuals (9.0 ± 2.7 pmol/L, all P < 0.001). Multivariate logistic regression analysis showed that plasma copeptin levels were independently related to CAP-related complications (odds ratio 1.214, 95% confidence interval 1.104–1.872, P < 0.001) and pleural effusion (odds ratio 1.226, 95% confidence interval 1.109–1.917, P < 0.001). A receiver operating characteristic curve analysis showed plasma copeptin level better predicted CAP-related complications (area under curve 0.876, 95% confidence interval 0.815–0.922) and pleural effusion (area under curve 0.831, 95% confidence interval 0.765–0.885). Thus, plasma copeptin level may represent a novel biomarker for predicting CAP-related complications in preschool children.     

Effects of dietary nitrate supplementation on the oxygen cost of exercise and walking performance in individuals with type 2 diabetes: a randomized,double-blind,placebo-controlled crossover trial

Dietary nitrate supplementation has been shown to reduce the oxygen (O₂) cost of exercise and enhance exercise tolerance in healthy individuals. This study assessed whether similar effects could be observed in individuals with type 2 diabetes (T2DM). In a randomized, double-blind, placebo-controlled crossover study, 48 participants with T2DM supplemented their diet for 4 days with either nitrate-rich beetroot juice (70 ml/day, 6.43 mmol nitrate/day) or nitrate-depleted beetroot juice as placebo (70 ml/day, 0.07 mmol nitrate/day). After each intervention period, resting plasma nitrate and nitrite concentrations were measured subsequent to participants completing moderate-paced walking. Pulmonary gas exchange was measured to assess the O₂ cost of walking. After a rest period, participants performed the 6-min walk test (6MWT). Relative to placebo, beetroot juice resulted in a significant increase in plasma nitrate (placebo, 57±66 vs beetroot, 319±110 µM; P < 0.001) and plasma nitrite concentration (placebo, 680±256 vs beetroot, 1065±607 nM; P < 0.001). There were no differences between placebo juice and beetroot juice for the O₂ cost of walking (946±221 vs 939±223 ml/min, respectively; P = 0.59) and distance covered in the 6MWT (550±83 vs 554±90 m, respectively; P = 0.17). Nitrate supplementation did not affect the O₂ cost of moderate-paced walking or improve performance in the 6MWT. These findings indicate that dietary nitrate supplementation does not modulate the response to exercise in individuals with T2DM.     

Differences in scapular kinematics and scapulohumeral rhythm during elevation and lowering of the arm between typical children and healthy adults

Scapular kinematics in healthy adults is well described in the literature but little is known on typical children. This study aimed to compare the three-dimensional (3-D) scapular kinematics and scapulohumeral rhythm during the elevation and lowering of the arm in the scapular plane in typical children and healthy adults. Twenty-six healthy adults (35.34 ± 11.65 years, 1.70 ± 0.10 m, 70.00 ± 12.30 kg) and 33 typical children (9.12 ± 1.51 years, 1.40 ± 0.10 m, 35.40 ± 10.45 kg) participated in this study. 3-D scapular kinematics were obtained using an electromagnetic tracking device. The subjects were asked to elevate and lower their arm in the scapular plane. Children showed less scapular protraction compared to adults at 120° during arm elevation, more anterior tilt than adults in the elevation and also at 60°, 90° and 120° during lowering of the arm. Children also showed higher scapulohumeral rhythm during lowering of the arm compared to adults from 90° to 60°. It was also found a low to little correlation between scapular position and age. The study showed small but significant differences in scapular kinematics and scapulohumeral rhythm between children and adults. These results can help clinicians to improve diagnosis and treatment protocols directed to children with dysfunction, as reference values on scapular kinematics in healthy children are also provided in this study.