Higher somatic cells counted by the electronic counter method do not influence renneting properties of goat milk

Laws of different countries regarding SCC of goat milk are not in agreement with each other and sometimes they fix a threshold for the enhancement of dairy products. The aim of this study was to assess if renneting properties of goat milk are influenced by higher somatic cell count (SCC) measured by an electronic cell counter. Milk samples, taken throughout the lactation of 169 goats from three farms, were analyzed for chemical, physical, hygienic and renneting properties. Samples were divided into three levels on the basis of their SCC: L: low level, samples with SCC lower than 10⁶ cells/ml; M: medium, between 10⁶ and 2 × 10⁶ cells/ml; H: high, higher than 2 × 10⁶ cells/ml. Milk clotting time was between 12.07 and 13.31 min, curd firming time between 1.68 and 2.05 min and curd firmness between 41.66 and 48.97 mm. All the three renneting properties were not affected by the SCC level but they were highly correlated with other factors as protein content and pH. Furthermore, the microbial count showed a high positive correlation with SCC. These results showed that in goat milk, contrarily to other dairy species, higher SCC did not affect renneting properties and that counting of somatic cells by using electronic cell counters might be not suitable for the improvement of dairy products.     

Presence of Mycoplasma species and somatic cell counts in bulk-tank goat milk

This cross-sectional study performed on dairy goat herds was designed to establish the relationship between the presence of Mycoplasma species in bulk-tank milk samples from different farms and the bulk-tank milk somatic cell count (BTMSCC) in an area where contagious agalactia (CA) is endemic. Three BTMSCC thresholds, used in payment schemes or as legal requirements for milk quality in Europe and the USA, were considered: (1) 2,000,000 cells/ml; (2) 1,500,000 cells/ml and (3) 1,000,000 cells/ml. Of the 1068 milk samples tested, 7.9% (n = 84) showed the presence of Mycoplasma spp. (Mycoplasma agalactiae 82% and Mycoplasma mycoides subsp. mycoides large colony 17%). Somatic cell counts for bulk-tank samples containing mycoplasmas were higher than those recorded for negative samples (1,176,000 cells/ml vs. 875,000 cells/ml; P < 0.001). Two-by-two table analyses revealed that the presence of mycoplasmas in bulk-tank milk increased the risk of surpassing all SCC thresholds considered, with the highest risk for Mycoplasma positive bulk-tank milk samples exceeding the threshold of 1500 × 10³ cells/ml (odds ratio = 2.42 (1.49 < OR < 3.91). Our results indicate that the presence of mycoplasmas in goat milk had yet another economic consequence and a further incentive to encourage the implementation of specific programs for disease control.     

Use of ethidium bromide monoazide for quantification of viable and dead mixed bacterial flora from fish fillets by polymerase chain reaction

Ethidium bromide monoazide (EMA) was utilized to selectively allow conventional PCR amplification of target DNA from viable but not dead cells from a broth culture of bacterial mixed flora derived from cod fillets. The universal primers designated DG74 and RW01 that amplify a 370-bp sequence of a highly conserved region of all eubacterial 16S rDNA were used for the PCR. The use of 10 μg/ml or less of EMA did not inhibit the PCR amplification of DNA derived from viable bacteria. The minimum amount of EMA to completely inhibit the PCR amplification of DNA derived from dead bacterial cells was 0.8 μg/ml. Amplification of target DNA from only viable cells in a suspension with dead cells was selectively accomplished by first treating the cells with 1 μg/ml of EMA. A standard curve was generated relating the intensity of fluorescence of DNA bands to the log of CFU of mixed bacterial cultures for rapidly assessing the number of genomic targets per PCR derived from the number of CFU. A linear range of DNA amplification was exhibited from 1 × 10² to 1 × 10⁵ genomic targets per PCR. The viable/dead cell discrimination with the EMA-PCR method was evaluated by comparison with plate counts following freezing and thawing. Thawing frozen cell suspensions initially containing 1 × 10⁵ CFU/ml at 4, 20, and 37 °C yielded a 0.8 log reduction in the number of viable cells determined by both plate counts and EMA-PCR. In contrast, thawing for 5 min at 70 °C resulted in a 5 log reduction in CFU derived from plate counts (no CFU detected) whereas the EMA-PCR procedure resulted in only a 2.8 log reduction in genomic targets, possibly reflecting greater damage to enzymes or ribosomes at 70 °C to a minority of the mixed population compared to membrane damage.     

Analysis of spatial and temporal variation in the levels of microbial fecal indicators in the major rivers flowing into the Aegean Sea,Turkey

River watersheds in the western part of Turkey (Aegean Sea) are anthropogenically impacted areas, due to the high population density, intensive agricultural and/or industrial activities. The aim of the present survey was to assess the microbiological quality of river waters using indicators of fecal contamination. Five rivers (Meric, Bakircay, Gediz, Kucuk Menderes and Buyuk Menderes) were sampled as seasonally from 2006 to 2008 for fecal coliforms, and fecal streptococci. In order to determine the number of fecal indicator bacteria, membrane filtration method was used. The minimum fecal coliforms and fecal streptococci were counted from the samples taken from the Buyuk Menderes River (5 × 10¹–3 × 10¹ CFU/100 ml) especially in the spring and autumn. The highest fecal coliform result was obtained in winters (1.3 × 10⁶ CFU/100 ml) while maximum fecal streptococci value was detected in autumns (6.3 × 10⁴ CFU/100 ml) in the Kucuk Menderes River during 2006–2008. Additionally, the relationships between the fecal indicator bacteria and physico-chemical parameters (temperature, conductivity and pH) were also evaluated, but no significant relationships were observed between the fecal indicator bacteria and environmental parameters. Considering the findings of these studies, it can be said that there is fecal pollution in the rivers reaching the Aegean Sea. The results show that there are a great number of microbial pollution sources in the areas where the river passes through and thus, in order to implement strategies to improve water quality in these rivers, monitoring of these rivers should continue.     

Characteristics of Korean-Saanen goat milk caseins and somatic cell counts in comparison with Holstein cow milk counterparts

Characteristics of α- and β-casein fractions in the milk of Korean-Saanen goats were compared with those of Holstein cow milk using capillary electrophoresis (CE) analysis. The αs₁-CN content of the Saanen goat milk samples varied from 2.4% to 9.3% of total proteins. Total αs-CN content of the goat milk varied from 10.1% to 17.0%. Total β-CN content containing β₁-CN and the β₂-CN varied from 49.6% to 61.0% of total proteins. Average αs-CN to β-CN ratio of the Saanen goat milk from different farms was 0.24 ± 0.04, ranging from 0.17 to 0.33. The αs-CN (αs₁-CN + αs₀-CN) to β-CN (β_A1-CN + β_A2-CN) ratio of Holstein cow milk was 0.81, which was much higher than that of Korean-Saanen goat milk. The goat milk samples having more than 1.5 million cells/ml somatic cell counts (SCC) contained higher αs-CNs (P < 0.01) and lower β-CNs (P < 0.05) contents than milks with <1.5 million SCC. This resulted in a higher αs-CN to β-CN ratio (P < 0.01) in the milk with >1.5 million SCC.     

Isolation and identification of gastrointestinal microbiota from the short-nosed fruit bat Cynopterus brachyotis brachyotis

Studies on the microbial ecology of gut microbiota in bats are limited and such information is necessary in determining the ecological significance of these hosts. Short-nosed fruit bats (Cynopterus brachyotis brachyotis) are good candidates for microbiota studies given their close association with humans in urban areas. Thus, this study explores the gut microbiota of this species from Peninsular Malaysia by means of biochemical tests and 16S rRNA gene sequences analysis. The estimation of viable bacteria present in the stomach and intestine of C. b. brachyotis ranged from 3.06 × 10¹⁰ to 1.36 × 10¹⁵ CFU/ml for stomach fluid and 1.92 × 10¹⁰ to 6.10 × 10¹⁵ CFU/ml for intestinal fluid. A total of 34 isolates from the stomach and intestine of seven C. b. brachyotis were retrieved. A total of 16 species of bacteria from eight genera (Bacillus, Enterobacter, Enterococcus, Escherichia, Klebsiella, Pantoea, Pseudomonas and Serratia) were identified, Enterobacteriaceae being the most prevalent, contributing 12 out of 16 species isolated. Most isolates from the Family Enterobacteriaceae have been reported as pathogens to humans and wildlife. With the possibility of human wildlife transmission, the findings of this study focus on the importance of bats as reservoirs of potential bacterial pathogens.     

Involvement and interaction of microbial communities in the transformation and stabilization of chromium during the composting of tannery effluent treated biomass of Vallisneria spiralis L.

Tannery effluent treated with aquatic macrophyte Vallisneria spiralis L. for 14 d showed significant improvement in physico-chemical properties and reduction in Cr concentration. Accumulation of Cr was found maximum in roots (358 μg g^?1dw) as compared to shoot (62 μg g^?1dw) of the plant. A laboratory scale composter was designed with the objectives to investigate the physico-chemical changes and role of microbes in stabilization and transformation of Cr in the composting material. Results revealed that the composting process was quick within 7–21 d as indicated by peak time for various physico-chemical parameters and drop in C/N ratio up to acceptable limit. The profile of microbial communities indicated that population of anaerobic, aerobic and nitrifying bacteria increased quickly at the initial phase, and reached a peak level of 4.2 × 10⁶, 9.78 × 10⁸ and 9.32 × 10⁹ CFU g^?1, respectively at 21 d; while population of actinomycetes and fungi was found maximum i.e. 3.29 × 10⁷ and 9.7 × 10⁶ CFU g^?1, respectively, after 35 d of composting. Overall bacterial population dominated over the actinomycetes and fungi during the composting process. Cr^(VI) was transformed to Cr^(III) due to the microbial activity during the process. Sequential extraction of Cr fractionation showed its stabilization via changing into organic matter-bound and residual fractions during the composting.     

Efficacy assessment of Pichia guilliermondii strain Z1, a new biocontrol agent,against citrus blue mould in Morocco under the influence of temperature and relative humidity

The interactions of Penicillium italicum, which causes blue mould, and antagonistic yeast Pichia guilliermondii strain Z1 were examined in controlled environments, to determine the influence of relative humidity (RH) (45%, 75%, 85%, 98%, and 100%) and temperature (T) (5, 10, 15, 20, and 25 °C). All main effects and interactions were significant (P ? 0.05), with the exception of interactions RH×T and strain Z1 (BCA)×RH×T. In the pathogen control, the lesion diameter of blue mould developed under all environmental conditions but was the largest at a RH range between 98% and 100%, independent of the temperature. The efficacy of strain Z1 appeared to be independent of the environment and reduced disease incidence by more than 85% in all environmental conditions. Rapid colonization of the antagonistic yeast strain Z1 on citrus wounded sites was recorded during the first week at 5 °C. Colonization then stabilized at ±6.9 × 10⁶ CFU/ml for 30 days. This indicates that P. guilliermondii is able to adapt itself and colonize the wound sites prior to the arrival of the pathogen, displaying greater efficiency than when colonizing wounds after pathogen. The antagonist was capable of growing in low concentrations of orange juice (0.1–5%), with greatest growth at 5%. Applying strain Z1 (1 × 10⁸ CFU/ml) as a formulated product significantly reduced the incidence of infected fruits and the percentage of infected wounds relative to the pathogen control. Disease control with formulated product (45%) was slightly lower than that obtained with thiabendazole (20%) or strain Z1 culturable cells (25%). These results suggest that strain Z1 may be a useful BCA for control of blue mould under varying environmental conditions, and control may be enhanced by combining with other eco-friendly post-harvest treatments or improved formulation.     

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.     

Biocontrol of major postharvest pathogens on apple using Rhodotorula glutinis and its effects on postharvest quality parameters

The biocontrol activity of Rhodotorula glutinis on gray mold decay and blue mold decay of apple caused by Botrytis cinerea and Penicillium expansum, respectively, was investigated, as well as its effects on postharvest quality of apple fruits. The results show there was a significant negative correlation between concentrations of the yeast cells and the disease incidence of the pathogens. The higher concentration of the R. glutinis, the better effect of the biocontrol capacity. At concentrations of R. glutinis 1 × 10⁸ CFU ml^?1, the amount of gray mold decay was completely inhibited after 5 days incubation at 20 °C, after challenge with B. cinerea spores suspension of 1 × 10⁵ spores ml^?1; While the blue mold decay was completely inhibited at concentrations of 5 × 10⁸ CFU ml^?1, at challenged with P. expansum spores suspension of 5 × 10⁴ spores ml^?1_. These results demonstrated that the efficacy of R. glutinis in controlling of gray mold decay of apples was better than the efficacy of controlling blue mold. R. glutinis within inoculated wounds on apples increased in numbers at 20 °C from an initial level of 9.5 × 10⁵ CFU per wound to 2.24 × 10⁷ CFU at 20 °C after 1 day. The highest population of the yeast was recovered 4 days after inoculation, the yeast population in wounds increased by 56.9 times. After that, the population of the yeast began to decline very slowly. R. glutinis significantly reduced the incidence of natural infections on intact fruit from 75% in the control fruit to 28.3% after 5 days at 20 °C, and from 58.3 to 6.7% after 30 days at 4 °C followed by 4 days at 20 °C. R. glutinis treatment had no deleterious effect on quality parameters after 5 days at 20 °C or after 30 days at 4 °C followed by 4 days at 20 °C.     

Sources of variation contributing to production and quality attributes of Kyrgyz cashmere in Osh and Naryn provinces: Implications for industry development

We aimed to quantify the sources of variation contributing to the production and quality of cashmere produced in five districts in Osh and Naryn provinces of Kyrgyzstan. In early spring 2008 mid-side cashmere samples were taken from 719 cashmere adult females, and 41 cashmere adult males and castrates. Samples came from 53 villages and a total of 156 farmers’ flocks. For 91 goats from 33 farmers in 13 villages of two districts that had been sampled earlier, cashmere was combed from the goat at the time of a second visit (end of April 2008) when the cashmere would normally be harvested. Following standard cashmere objective measurement, data were examined using general linear modelling to quantify the effects of potential determinants. The mean fibre diameter (MFD) of cashmere differed between provinces (Osh 15.7 μm, Naryn 16.7 μm; P = 4.4 × 10⁻²⁰). About 42% of the cashmere was <16 μm, 48% was 16.0–18.0 μm and 9.5% was >18.0 μm. Most of the cashmere samples were coloured (81%), with 63% black and 19% white. The percentage of cashmere samples that were white declined as MFD increased (26% < 14 μm to 11% of >18 μm). The primary determinants of cashmere MFD of individual goats were age of goat (range 1.46 μm, P = 1.8 × 10⁻¹²) and farm (range 6.5 μm, P = 1.7 × 10⁻¹⁴). The lesser effects detected for sex (range 0.9 μm, P = 0.026) and colour of cashmere (range 1.8 μm, P = 0.023) were based on small sample sizes and are unreliable. Age of goat had important affects on fibre diameter variation (up to 1.7% in coefficient of variation, P = 5.8 × 10⁻⁶) and fibre curvature (2.5–5°/mm, P = 2.1 × 10⁻⁴). By far the greatest effect on fibre curvature was cashmere MFD (P = 3.0 × 10⁻¹⁰⁴) with a smaller effect of sex (about 5°/mm, P = 3.0 × 10⁻⁶). Village effects were detected on fibre diameter variability (range 4.5% in coefficient of variation, P = 0.027) and fibre curvature (range 15°/mm, P = 1.6 × 10⁻⁷). There was a strong negative association between increasing MFD and declining fibre curvature (−5.11 ± 0.181°/mm per 1 μm; P = 7.1 × 10⁻¹²¹; r² = 0.51). Average combed cashmere weight was 164 g, the clean cashmere content was 0.661 and median clean cashmere production was 110 g per goat (range 60–351 g). Combed cashmere production increased with altitude of the village, probably related to different moulting times as spring temperatures warmed up later in higher altitude villages up to 3200 masl. Measurements of combed cashmere MFD were coarser than the mid-side samples taken earlier in the year. There are farmers and cashmere goats in the sampled districts of Kyrgyzstan which produce the finest qualities of commercial cashmere as the vast majority of cashmere is fine, has low variation in fibre diameter and has fibre crimping (curvature) typical of Chinese and Mongolian cashmere. There is substantial scope to increase the production and commercial value of cashmere produced by Kyrgyz goats. In particular, some villages and farmers need to change their buck selection practices if they wish to produce acceptable cashmere. Farmers should separate their finer and white cashmere prior to sale.     

Identification of lactic acid bacteria and yeast from boza

Boza is a low-alcohol beverage produced from the fermentation of barley, oats, millet, maize, wheat or rice. The number of lactic acid bacteria isolated from three boza samples ranged from 9 × 10⁶ to 5 × 10⁷ CFU/mL. Carbohydrate fermentation reactions and PCR with species-specific primers classified the isolates as Lactobacillus paracasei subsp. paracasei, Lactobacillus pentosus, Lactobacillus plantarum, Lactobacillus brevis, Lactobacillus rhamnosus and Lactobacillus fermentum. No filamentous fungi were isolated. Yeasts were isolated from two of the three boza samples, with cell numbers ranging from 1.3 × 10² to 1.8 × 10³ CFU/mL. Results obtained from sequencing of the D1/D2 rDNA region identified the yeasts as Candida diversa, Candida inconspicua, Candida pararugosa, Issatchenkia orientalis, Pichia fermentans, Pichia guillliermondii, Pichia norvegensis, Rhodotorula mucilaginosa and Torulaspora delbrueckii. C. inconspicua has been isolated from human sputum and tongue and is an opportunistic pathogen. R. mucilaginosa is also an opportunistic pathogen implicated in fungaemia, endocarditis and meningitis. P. norvegensis has been associated with septicaemia in humans. Saccharomyces cerevisiae, commonly associated with fermented beverages, has not been detected in any of the boza samples, despite enrichment.     

Immobilization of acidic lipase derived from Pseudomonas gessardii onto mesoporous activated carbon for the hydrolysis of olive oil

Mesoporous activated carbon (MAC) derived from rice husk is used for the immobilization of acidic lipase (ALIP) produced from Pseudomonas gessardii. The purified acidic lipase had the specific activity and molecular weight of 1473 U/mg and 94 kDa respectively. To determine the optimum conditions for the immobilization of lipase onto MAC, the experiments were carried out by varying the time (10–180 min), pH (2–8), temperature (10–50 °C) and the initial lipase activity (49 × 10³, 98 × 10³, 147 × 10³ and 196 × 10³ U/l in acetate buffer). The optimum conditions for immobilization of acidic lipase were found to be: time—120 min; pH 3.5; temperature—30 °C, which resulted in achieving a maximum immobilization of 1834 U/g. The thermal stability of the immobilized lipase was comparatively higher than that in its free form. The free and immobilized enzyme kinetic parameters (K_m and V_max) were found using Michaelis–Menten enzyme kinetics. The K_m values for free enzyme and immobilized one were 0.655 and 0.243 mM respectively. The immobilization of acidic lipase onto MAC was confirmed using Fourier Transform-Infrared Spectroscopy, X-ray diffraction analysis and scanning electron microscopy.     

A primary Corynebacterium pseudotuberculosis low dose infection in alpacas (Lama pacos) protects against a lethal challenge exposure

Corynebacterium pseudotuberculosis is the agent of alpaca's lymphadenitis. The present study was to demonstrate the effect of a primary infection with low (1.1 × 10³), moderate (1 × 10⁴), and high (1.2 × 10⁵) doses of C. pseudotuberculosis against a significant higher challenge dose of 9 × 10⁸ CFU of C. pseudotuberculosis. Three groups of 4 healthy male alpacas were inoculated subcutaneously (SC) in the left flank behind the costal arch with the above doses of bacteria. A fourth group of 4 alpacas was sham inoculated with phosphate buffered saline as control. After 5 weeks all animals were challenged with a dose of 9 × 10⁸ CFU of C. pseudotuberculosis inoculated SC in the right flank. The alpacas were clinically inspected for local and regional abscesses, body temperature and behavior changes. The primary infected alpacas had a febrile response, and abscesses at the inoculation point and regional lymph nodes. However, after challenge, the primary infected animals showed no superficial lesions or febrile response. In contrast, the immune naïve alpacas from group D developed a severe disease characterized by fever, abscesses in regional lymphnodes, and in one alpaca a subcutaneous edema and sudden death 2 weeks after exposure. In addition, primary infected alpacas had a robust antibody response against C. pseudotuberculosis cell wall antigen with significant differences with respect the naïve challenged alpacas. At necropsy, the primary infected alpacas had abscesses only in the regional or internal renal-lymph nodes from the left or primary inoculation side of the body, with no lesions in the right challenged side. In contrast, the primary sham inoculated alpacas had abscesses in the regional and internal lymph nodes from the right challenged side. This work showed that a primary infection with at least 1.1 × 10³ viable C. pseudotuberculosis induces protection against a second high dose exposure to this bacterium. These results will be useful for further study of prevention methods to control lymphadenitis in alpacas.     

Differentiating between isolates of Vibrio vulnificus with monoclonal antibodies

Monoclonal antibodies (MAbs) against Vibrio vulnificus (isolate I, VVC and isolate II, VVB) were raised using heat-killed and heat-killed plus SDS–mercaptoethanol treated forms of VVC and VVB for immunizing Swiss mice. Twenty three hybridomas producing MAbs against V. vulnificus were selected and divided into five groups according to their specificities to different V. vulnificus isolates and apparent protein antigens which ranged from ∼ 3–50 kDa. Four groups were specific to V. vulnificus without cross reactivity to either other Vibrio spp. or other bacterial species. In dot blot based assays, one group of MAbs were specific to VVC, with a sensitivity of ∼ 1.6 × 10⁷ CFU ml^− 1 (∼ 1.6 × 10⁴ cells spot^− 1), and bound to proteins of ∼ 50 and ∼ 39 kDa. Other MAbs, binding to proteins ranging from ∼ 3–14 and ∼ 40 kDa, detected VVB (but not VVC) with high sensitivity at ∼ 1.6 × 10⁵ and 4 × 10⁶ CFU ml^− 1 (∼ 1.6 × 10² and 4 × 10³ cells spot^− 1), respectively. In addition, certain MAbs were able to recognize V. vulnificus in tissues by means of immunohistochemistry. The remaining groups demonstrated cross reactivity to Vibrio fluvialis. MAbs from this study can, therefore, detect the difference between some isolates of V. vulnificus and in addition to pathogen detection may, with further antibodies, form the basis of serovar typing isolates in the future.     

Inactivation of IL-6 and soluble IL-6 receptor by neutrophil derived serine proteases in cystic fibrosis

The ability of IL-6 to signal via both membrane bound and soluble receptors is thought to explain the capacity of this cytokine to act in both the initiation and resolution of acute inflammatory responses. In cystic fibrosis (CF), poorly resolved neutrophillic inflammation of the lungs is a primary cause of morbidity and mortality. Expression of IL-6 has been reported to be low in CF lung secretions, despite ongoing inflammation, but the status of soluble IL-6 receptor (sIL-6R) in these patients is unknown. We hypothesised that sIL-6R may be an important potentiator of IL-6 activity in CF associated lung disease. IL-6, sIL-6R and sgp130 (a natural antagonist of responses mediated by the sIL-6R) were analysed by ELISA and Western blot in bronchoalveolar lavage fluid (BALF) from 28 paediatric CF patients and nine non-CF controls. Total cell counts in CF were four fold higher compared to controls (median: 1.4 × 10⁶ cells/ml v. 0.35 × 10⁶ cells/ml in controls) (p < 0.001) and the infiltrate was dominated by neutrophils which were elevated by 89 fold (0.62 × 10⁶ cells/ml v. 0.007 × 10⁶ cells/ml in controls) (p < 0.001). Other markers of inflammation such as IL-8 and MCP-1 were elevated 17.5 and 3.8 fold respectively (IL-8; median: 1122 pg/ml v. 64 pg/ml in controls, p < 0.01 and MCP-1; median: 692 pg/ml v. 182 pg/ml in controls, p < 0.05). IL-6, although present in 23/32 CF BALF specimens compared to 1/9 controls (p < 0.01), was weakly expressed (median: 50 pg/ml). Expression of sIL-6R and sgp130 in CF was no different to control patients. We tested whether weak expression of all three molecules was due to degradation by CF BALF. Degradative activity was observed in association with BALF elastase activity and could be specifically blocked by serine protease inhibitors. Degradation of sIL-6R by purified serine proteases (elastase, cathepsin G and proteinase 3) was also observed leading to a loss of trans-signalling activity. Interestingly, sIL-6R was protected from proteolysis by interaction with IL-6. Our data identify and define a novel protease mediated deficiency of IL-6 signalling in the CF lung.     

Intestinal and immunological effects of daily oral administration of Lactobacillus salivarius CECT5713 to healthy adults

There is an increasing interest in the intestinal and immunological effects of probiotics. The aim of the present study is to evaluate the tolerance and beneficial effects in healthy adults of the strain, Lactobacillus salivarius CECT5713 isolated from breast milk. A phase II, randomized, double-blinded, placebo-controlled human clinical trial was carried out in 40 healthy adults. The Probiotic group received a daily dose of 2 × 10⁸ CFU of L. salivarius CECT5713 in capsules during 4 weeks while volunteers of the control received only a placebo. Gastrointestinal and immunological parameters were analyzed. Results showed that L. salivarius CECT5713 was well tolerated and no adverse effects were detected. Consumption of the probiotic strain increased fecal lactobacilli counts (7.9 ± 0.1 vs. 7.05 ± 0.2 CFU/g feces, P = 0.001). Also, an improvement in the frequency of defecation (P = 0.04) was observed. Probiotic treatment induced significantly the percentage of NK cells and monocytes, as well as the plasmatic levels of immunoglobulins M, A and G, and the regulatory cytokine IL-10 (72.3 ± 11.7 in probiotic group vs. 27.3 ± 6.4 pg/mL in control group, P < 0.01). Thus, it can be concluded that daily administration of L. salivarius CECT5713 to healthy adults is safe and improve gut microbiota and different parameters related to immune response.     

Clinical significance of serum transforming growth factor-β1 in lung cancer

Background: Transforming growth factor-β1 (TGF-β1) plays a critical role in human cancer development. Present study aimed to explore the clinical significance of serum TGF-β1 levels in patients with lung cancer and analyze the relationship between TGF-β1 and existing tumor markers for lung cancer. Methods: Serum was collected from 118 patients with lung cancer and 40 healthy volunteers. Serum TGF-β1 levels were measured by enzyme-linked immunosorbent assay (ELISA), and the association with various clinical characteristics was analyzed. The diagnostic value of TGF-β1 was assessed alone and in combination with existing tumor markers for lung cancer. Results: Serum TGF-β1 levels were significantly higher in patients with lung cancer compared to healthy volunteers [0.6 × 10⁵ (0.4 × 10⁵, 0.9 × 10⁵) pg/ml vs 0.5 × 10⁵ (0.3 × 10⁵, 0.7 × 10⁵) pg/ml, P = 0.040]. Although there was a positive correlation between serum TGF-β1 levels and advanced stages, the significant difference was not found between early stages and advanced stages (P = 0.116). The ability of serum TGF-β1 to discriminate lung cancer at a cutoff value of 79,168 pg/ml exhibited sensitivity of 30.6% and specificity of 97.5%. Serum TGF-β1 levels were correlated to cytokeratin fragment 21-1 (CYFRA21-1; R = 0.308, P = 0.020) and neuron-specific enolase (NSE; R = 0.558, P = 0.003). The diagnostic accuracy rates for the existing lung-tumor markers, as SCC, CYFRA21-1, and NSE, were increased from 20.0%, 34.6%, and 45.9% to 48.9%, 51.7%, and 54.5%, respectively by the inclusion of serum TGF-β1 levels. Conclusion: Quantification of serum TGF-β1 levels by ELISA may provide a novel complementary tool for the clinical diagnosis of lung cancer.     

Immobilization of alkaline serine endopeptidase from Bacillus licheniformis on SBA-15 and MCF by surface covalent binding

An industrial enzyme, alkaline serine endopeptidase, was immobilized on surface modified SBA-15 and MCF materials by amide bond formation using carbodiimide as a coupling agent. The specific activities of free enzyme and enzyme immobilized on SBA-15 and MCF were studied using casein (soluble milk protein) as a substrate. The highest activity of free enzyme was obtained at pH 9.5 while this value shifted to pH 10 for SBA-15 and MCF immobilized enzyme. The highest activity of immobilized enzymes was obtained at higher temperature (60 °C) than that of the free enzyme (55 °C). Kinetic parameters, Michaelis–Menten constant (K_m) and maximum reaction velocity (V_max), were calculated as K_m = 13.375, 11.956, and 8.698 × 10^?4 mg/ml and V_max = 0.156, 0.163 and 0.17 × 10^?3 U/mg for the free enzyme and enzyme immobilized on SBA-15 and MCF, respectively. The reusability of immobilized enzyme showed 80% of the activity retained even after 15 cycles. Large pore sized MCF immobilized enzyme was found to be more promising than the SBA-15 immobilized enzyme due to the availability of larger pores of MCF, which offer facile diffusion of substrate and product molecules.     

Preslaughter spray-washing effects on physiological stress responses and skin and carcass microbial counts in goats

Preslaughter management procedures that decrease fecal contamination of skins/hides are likely to reduce biological hazards on carcass surfaces during slaughter and processing. This trial was conducted to determine the effects of preslaughter spray-washing on stress responses and skin and carcass bacterial counts in goats. Twenty meat goats were slaughtered in two groups (replicate) on 2 different days (10 goats/replicate). Animals were randomly allotted to treatment (1 min spray-wash) or control (no wash) groups (n = 5/(treatment replicate)). Blood and skin swab samples were collected from control and treated animals before and after the washing treatment. Bacterial counts on the carcasses immediately after dressing were also recorded. Treatment, sampling time or treatment × sampling time did not affect the plasma cortisol, glucose and non-esterified fatty acid (NEFA) concentrations. Skin aerobic plate counts were the same in both treated and control groups prior to washing treatment, but were significantly less in the treated group when sampled after washing (treatment × sampling time, P < 0.05). Aerobic plate counts were 3.6 and 4.4 log₁₀ CFU/cm² in the treated and control groups, respectively. However, skin Escherichia coli counts did not significantly decrease due to spray-washing treatment. Spray-washing treatment also did not influence carcass E. coli or aerobic plate counts. Results indicate that skin bacterial counts can be significantly reduced by preslaughter spray-washing, without increasing stress in goats. Preslaughter spray-washing may be a cost-effective skin decontamination method that can be easily adopted in goat slaughter plants.