Phenotypic and Genotypic Characterization of Bacteriocinogenic Enterococci Against <Emphasis Type="Italic">Clostridium botulinum</Emphasis>

The present study aimed to characterize Enterococcus faecalis (n = ?6) and Enterococcus faecium (n = 1) isolated from healthy chickens to find a novel perspective probiotic candidate that antagonize Clostridium botulinum types A, B, D, and E. The isolated enterococci were characterized based on phenotypic properties, PCR, and matrix-assisted laser desorption/ionization time of flight (MALDI-TOF). The virulence determinants including hemolytic activity on blood agar, gelatinase activity, sensitivity to vancomycin, and presence of gelatinase (gelE) and enterococcal surface protein (esp) virulence genes were investigated. Also, the presence of enterocin structural genes enterocin A, enterocin B, enterocin P, enterocin L50A/B, bacteriocin 31, enterocin AS48, enterocin 1071A/1071B, and enterocin 96 were assessed using PCR. Lastly, the antagonistic effect of the selected Enterococcus spp. on the growth of C. botulinum types A, B, D, and E was studied. The obtained results showed that four out of six E. faecalis and one E. faecium proved to be free from the tested virulence markers. All tested enterococci strains exhibited more than one of the tested enterocin. Interestingly, E. faecalis and E. faecium significantly restrained the growth of C. botulinum types A, B, D, and E. In conclusion, although, the data presented showed that bacteriocinogenic Enterococcus strains lacking of virulence determinants could be potentially used as a probiotic candidate against C. botulinum in vitro; however, further investigations are still urgently required to verify the beneficial effects of the tested Enterococcus spp. in vivo.     

A newly isolated and identified vitamin B<Subscript>12</Subscript> producing strain: <Emphasis Type="Italic">Sinorhizobium meliloti</Emphasis> 320

Vitamin B₁₂ (Cobalamin, VB₁₂) has several physiological functions and is widely used in pharmaceutical and food industries. A new unicellular species was extracted from China farmland, and the strain could produce VB₁₂ which was identified by HPLC and HPLC–MS/MS. 16S rDNA analysis reveals this strain belongs to the species Sinorhizobium meliloti and we named it S. meliloti 320. Its whole genome information indicates that this strain has a complete VB₁₂ synthetic pathway, which paves the way for further metabolic engineering studies. The optimal carbon and nitrogen sources are sucrose and corn steep liquor (CSL) plus peptone. The optimal combination of sucrose and CSL was obtained by response surface methodology as they are the most suitable carbon and nitrogen sources, respectively. This strain could produce 140 ± 4.2 mg L^?1 vitamin B₁₂ after incubating for 7 days in the optimal medium.     

Impact of elevated CO<Subscript>2</Subscript> in <Emphasis Type="Italic">Casuarina equisetifolia</Emphasis> rooted stem cuttings inoculated with <Emphasis Type="Italic">Frankia</Emphasis>

Impact of different levels of elevated CO ₂ on the activity of Frankia (Nitrogen-fixing actinomycete) in Casuarina equisetifolia rooted stem cuttings has been studied to understand the relationship between C. equisetifolia, Frankia and CO₂. The stem cuttings of C. equietifolia were collected and treated with 2000 ppm of Indole Butyric Acid (IBA) for rooting. Thus vegetative propagated rooted stem cuttings of C. equisetifolia were inoculated with Frankia and placed in the Open top chambers (OTC) with elevated CO₂ facilities. These planting stocks were maintained in the OTC for 12 months under different levels of elevated CO₂ (ambient control, 600 ppm, 900 ppm). After 12 months, the nodule numbers, bio mass, growth, and photosynthesis of C. equisetifolia rooted stem cuttings inoculated with Frankia were improved under 600 ppm of CO₂. The rooted stem cuttings of C. equisetifolia inoculated with Frankia showed a higher number of nodules under 900 ppm of CO₂ and cuttings without Frankia inoculation exhibited poor growth. Tissue Nitrogen (N) content was also higher under 900 ppm of CO₂ than ambient control and 600 ppm levels. The photosynthetic rate was higher (17.8 μ mol CO₂ m^?2 s^?1) in 900 ppm of CO₂ than in 600 ppm (13.2 μ mol CO₂ m^?2 s^?1) and ambient control (8.3 μ mol CO₂ m^?2 s^?1). This study showed that Frankia can improve growth, N fixation and photosynthesis of C. equietifolia rooted stem cuttings under extreme elevated CO₂ level conditions (900 ppm).     

Magnetophoretic harvesting of freshwater microalgae using polypyrrole/Fe<Subscript>3</Subscript>O<Subscript>4</Subscript> nanocomposite and its reusability

A magnetophoretic harvesting agent, a polypyrrole/Fe₃O₄ magnetic nanocomposite, is proposed as a cost and energy efficient alternative to recover biomass of the microalgae Botryococcus braunii, Chlorella protothecoides, and Chlorella vulgaris from their culture media. The maximal recovery efficiency reached almost 99 % for B. braunii, 92.4 % for C. protothecoides, and 90.8 % for C. vulgaris. The maximum adsorption capacity (Q ₀) of the magnetic nanocomposite for B. braunii (63.49 mg dry biomass mg^?1 PPy/Fe₃O₄) was higher than that for C. protothecoides (43.91 mg dry biomass mg^?1 PPy/Fe₃O₄) and C. vulgaris (39.98 mg dry biomass mg^?1 PPy/Fe₃O₄). The highest harvesting efficiency for all the studied microalgae were at pH 10.0, and measurement of zeta-potential confirmed that the flocculation was induced by charge neutralization. This study showed that polypyrrole/Fe₃O₄ can be a promising flocculant due to its high efficacy, low dose requirements, short settling time, its integrity with cells, and with great potential for saving energy because of its recyclability.     

Effect of <Emphasis Type="Italic">Lactobacillus reuteri</Emphasis> on Cell Viability and PGE<Subscript>2</Subscript> Production in Human Gingival Fibroblasts

Emerging evidence suggests that probiotic therapy can play a role in the prevention and management of oral inflammatory diseases through immunomodulation and down-regulation of the inflammatory cascade. The aim of this in vitro study was to investigate the viability of human gingival fibroblasts (HGF) and its production of prostaglandin E₂ (PGE₂), when exposed to supernatants of two mixed Lactobacillus reuteri strains (ATCC PTA 5289 and DSM 17938). The experiments were conducted in the presence and absence of the pro-inflammatory cytokine IL-1β. L. reuteri strains were grown and the bacterial supernatant was collected. The cell-free supernatant was diluted to concentrations equivalent to the ones produced by 0.5 to 5.0 × 10⁷ CFU/mL bacteria. Cell viability was assessed with the MTT colorimetric assay and the amount of PGE₂ in the cell culture medium was determined using the monoclonal enzyme immune assay kits. Our findings showed that none of the L. reuteri supernatants were cytotoxic or affected the viability of HGF. The most concentrated bacterial supernatant stimulated the production of PGE₂ by the gingival cells in a significant way in the presence of IL-1β (p < 0.05), suggesting that bacterial products secreted from L. reuteri might play a role in the resolution of inflammation in HGF. Thus, our findings justify further investigations on the influence of probiotic bacteria on gingival inflammatory reactions.     

Dielectric Dispersion in Ga<Subscript>2</Subscript>S<Subscript>3</Subscript> Thin Films

In this work, the structural, compositional, optical, and dielectric properties of Ga₂S₃ thin films are investigated by means of X-ray diffraction, scanning electron microscopy, energy dispersion X-ray analysis, and ultraviolet—visible light spectrophotometry. The Ga₂S₃ thin films which exhibited amorphous nature in its as grown form are observed to be generally composed of 40.7 % Ga and 59.3 % S atomic content. The direct allowed transitions optical energy bandgap is found to be 2.96 eV. On the other hand, the modeling of the dielectric spectra in the frequency range of 270–1,000 THz, using the modified Drude-Lorentz model for electron-plasmon interactions revealed the electrons scattering time as 1.8 (fs), the electron bounded plasma frequency as ~0.76–0.94 (GHz) and the reduced resonant frequency as 2.20–4.60 ×10¹⁵ (Hz) in the range of 270–753 THz. The corresponding drift mobility of electrons to the terahertz oscillating incident electric field is found to be 7.91 (cm ²/Vs). The values are promising as they nominate the Ga₂S₃ thin films as effective candidates in thin-film transistor and gas sensing technologies.     

Stability of fumonisin B<Subscript>1</Subscript>, deoxynivalenol,zearalenone, and T-2 toxin during processing of traditional Nigerian beer and spices

The stability of the Fusarium mycotoxins fumonisin B₁, deoxynivalenol, T-2 toxin, and zearalenone during processing of Nigerian traditional spices (dawadawa, okpehe, and ogiri) and beer (burukutu) using artificially contaminated raw materials was investigated. Results revealed the reduction of these toxins in all the final products. Boiling played a significant role (p?<?0.05) in Fusarium mycotoxin reduction in the traditional spices. The highest percentage reduction of deoxynivalenol (76%) and zearalenone (74%) was observed during okpehe processing (boiled for 12 h). Dehulling and fermentation further demonstrated a positive influence on the reduction of these toxins with a total reduction ranging from 85 to 98% for dawadawa, 86 to 100% for okpehe, and 57 to 81% for ogiri. This trend was also observed during the production of traditional beer (burukutu), with malting and brewing playing a major impact in observed reduction. In addition, other metabolites including deoxynivalenol-3-glucoside, 15-acetyl-deoxynivalenol, α-zearalenol, and β-zearalenol which were initially not present in the raw sorghum were detected in the final beer product at the following concentrations 26?±?11, 16?±?7.7, 22?±?18, and 31?±?16 μg/kg, respectively. HT-2 toxin was also detected at a concentration of 36?±?13 μg/kg along the processing chain (milled malted fraction) of the traditional beer. For the traditional spices, HT-2 toxin was detected (12 μg/kg) in ogiri. Although there was a reduction of mycotoxins during processing, appreciable concentrations of these toxins were still detected in the final products. Thus, the use of good quality raw materials significantly reduces mycotoxin contamination in final products.     

Molecular mapping of a rust resistance gene <Emphasis Type="Italic">R</Emphasis><Subscript><Emphasis Type="Italic">14</Emphasis></Subscript> in cultivated sunflower line PH 3

Sunflower, the fifth largest oilseed crop in the world, plays an important role in human diets. Recently, sunflower production in North America has suffered serious yield losses from newly evolved races of sunflower rust (Puccinia helianthi Schwein.). The rust resistance gene, designated R ₁₄ , in a germplasm line PH 3 originated from a wild Helianthus annuus L. population resistant to 11 rust races. PH 3 has seedling with an extraordinary purple hypocotyl color. The objectives of this study were to map both the R ₁₄ rust resistance gene and the purple hypocotyl gene-designated PHC in PH 3, and to identify molecular markers for marker-assisted breeding for sunflower rust resistance. A set of 517 mapped SSR/InDel and four SNP markers was used to detect polymorphisms between the parents. Fourteen markers covering a genetic distance of 17.0 cM on linkage group (LG) 11 were linked to R ₁₄ . R ₁₄ was mapped to the middle of the LG, with a dominant SNP marker NSA_000064 as the closest marker at a distance of 0.7 cM, and another codominant marker ORS542 linked at 3.5 cM proximally. One dominant marker ZVG53 was linked on the distal side at 6.9 cM. The PHC gene was also linked to R ₁₄ with a distance of 6.2 cM. Chi-squared analysis of the segregation ratios of R ₁₄ , PHC, and ten linked markers indicated a deviation from an expected 1:2:1 or 3:1 ratio. The closely linked molecular or morphological markers could facilitate sunflower rust-resistant breeding and accelerate the development of rust-resistant hybrids.     

Possible Probiotic Lactic Acid Bacteria Isolated from Oysters (<Emphasis Type="Italic">Crassostrea gigas</Emphasis>)

We attempted to isolate lactic acid bacteria (LAB) from the marine oyster (Crassostrea gigas) and selected several environmental stress-resistant isolates for the development of a future probiotic adjuvant for marine aquaculture. Twenty-six presumptive LAB isolates were extracted from oysters and screened (by an agar diffusion assay) for antimicrobial activity toward various pathogens: Vibrio parahaemolyticus, Streptococcus iniae, and Edwardsiella tarda. Eight isolates had an antibacterial activity toward V. parahaemolyticus; in particular, 6 isolates showed a growth-inhibitory activity, with inhibition zone diameters > 15 mm. Of these, 5 isolates (JL17, JL18, JL28, HL7, and HL32) were also active against S. iniae and E. tarda. Enterococcus faecium HL7 was selected as the isolate most resistant to environmental stressors: the minimum NaCl, ethanol, and hydrogen peroxide concentrations at which HL7 cells lost their viability were 1.9 M, 11%, and 0.013%, respectively. When an antibiotic sensitivity test was performed on E. faecium HL7, this isolate was found to be resistant to trimethoprim/sulfamethoxazole, cephalothin, ampicillin, rifampin, gentamicin, cefotaxime, cefepime, cefotetan, nalidixic acid, and kanamycin. While the oyster model studies provided indication that E. faecium HL7 could be a good candidate as biocontrol agent against V. vulnificus, further optimization is needed in the actual animal rearing situation.     

Interacting effects of elevated atmospheric CO<Subscript>2</Subscript> and hydrology on the growth and carbon sequestration of <Emphasis Type="Italic">Sphagnum</Emphasis> moss

Peatlands are a critical carbon store comprising 30% of the Earth’s terrestrial soil carbon. Sphagnum mosses comprise up to 90% of peat in the northern hemisphere but impacts of climate change on Sphagnum mosses are poorly understood, limiting development of sustainable peatland management and restoration. This study investigates the effects of elevated atmospheric CO₂ (eCO₂) (800 ppm) and hydrology on the growth of Sphagnum fallax, Sphagnum capillifolium and Sphagnum papillosum and greenhouse gas fluxes from moss–peat mesocosms. Elevated CO₂ levels increased Sphagnum height and dry weight but the magnitude of the response differed among species. The most responsive species, S. fallax, yielded the most biomass compared to S. papillosum and S. capillifolium. Water levels and the CO₂ treatment were found to interact, with the highest water level (1 cm below the surface) seeing the largest increase in dry weight under eCO₂ compared to ambient (400 ppm) concentrations. Initially, CO₂ flux rates were similar between CO₂ treatments. After week 9 there was a consistent three-fold increase of the CO₂ sink strength under eCO₂. At the end of the experiment, S. papillosum and S. fallax were greater sinks of CO₂ than S. capillifolium and the ? 7 cm water level treatment showed the strongest CO₂ sink strength. The mesocosms were net sources of CH₄ but the source strength varied with species, specifically S. fallax produced more CH₄ than S. papillosum and S. capillifolium. Our findings demonstrate the importance of species selection on the outcomes of peatland restoration with regards to Sphagnum’s growth and GHG exchange.     

Selection of Potential Probiotic <Emphasis Type="Italic">Lactobacillus</Emphasis> with Inhibitory Activity Against <Emphasis Type="Italic">Salmonella</Emphasis> and Fecal Coliform Bacteria

Three hundred and sixty presumptive lactic acid bacteria (LAB) isolated from pregnant sows, newborn, suckling, and weaned piglets were preliminarily screened for anti-Salmonella activity. Fifty-eight isolates consisting of Lactobacillus reuteri (n = 32), Lactobacillus salivarius (n = 10), Lactobacillus mucosae (n = 8), Lactobacillus johnsonii (n = 5), and Lactobacillus crispatus (n = 3) were selected and further characterized for probiotic properties including production of antimicrobial substances, acid and bile tolerance, and cell adherence to Caco-2 cells. Eight isolates including Lact. johnsonii LJ202 and Lact. reuteri LR108 were identified as potential probiotics. LJ202 was selected for further use in co-culture studies of two-bacterial and multiple-bacterial species to examine its inhibitory activity against Salmonella enterica serovar Enteritidis DMST7106 (SE7106). Co-culture of LJ202 and SE7106 showed that LJ202 could completely inhibit the growth of SE7106 in 10 h of co-culture. In co-culture of multiple-bacterial species, culturable fecal bacteria from pig feces were used as representative of multiple-bacterial species. The study was performed to examine whether interactions among multiple-bacterial species would influence antagonistic activity of LJ202 against SE7106 and fecal coliform bacteria. Co-culture of SE7106 with different combinations of fecal bacteria and probiotic (LJ202 and LR108) or non-probiotic (Lact. mucosae LM303) strains revealed that the growth of SE7106 was completely inhibited either in the presence or in the absence of probiotic strains. Intriguingly, LJ202 exhibited notable inhibitory activity against fecal coliform bacteria while LR108 did not. Taken together, the results of co-culture studies suggested that LJ202 is a good probiotic candidate for further study its inhibitory effects against pathogen infections in pigs.     

Occurrence and fumonisin B<Subscript>2</Subscript> producing potential of <Emphasis Type="Italic">Aspergillus</Emphasis> section <Emphasis Type="Italic">Nigri</Emphasis> in Brazil nuts

Bertholletia excelsa is the tree that produces Brazil nuts which have vast economic importance in the Amazon region and as an export commodity. The aim of this study was to assess the presence of Aspergillus section Nigri in Brazil nut samples at different stages of its production chain and to verify the toxigenic potential for fumonisin B₂ (FB₂) production of these isolates along with the presence of this mycotoxin in Brazil nut samples. The fungal infection ranged from 0 to 80% at the different stages of the harvest and processing chain and the water activity of the nuts from 0.273 to 0.994. A total of 1052 A. section Nigri strains were isolated from Brazil nuts and 200 strains were tested for their ability to produce FB₂: 41 strains (20.5%) were FB₂ producers with concentrations ranging from 0.09 to 37.25 mg/kg; 2 strains (1%) showed traces of FB₂, less than the detection limit (0.08 mg/kg); and 157 (78.5%) were not FB₂ producers. Although several samples showed high contamination by A. section Nigri, no sample was contaminated by FB₂.     

Biological Data on <Emphasis Type="Italic">Anovia punica</Emphasis> Gordon (Coleoptera: Coccinellidae), a Predator of <Emphasis Type="Italic">Crypticerya multicicatrices</Emphasis> Kondo &amp; Unruh (Hemiptera: Monophlebidae)

The coccinellid beetle Anovia punica Gordon (Coleoptera: Coccinellidae: Noviini) is an important predator of the Colombian fluted scale, Crypticerya multicicatrices Kondo & Unruh (Hemiptera: Monophlebidae). In order to gather information on the biological traits of A. punica, we conducted a series of studies, including of the developmental time of each life history stage, estimation of life table parameters, and predation rates under laboratory conditions [25.1 ± 1.6°C, with 70.5 ± 7.3% RH, and natural light regime, approx. 12:12 (L:D) h]. Developmental stages of A. punica were categorized as follows: egg stage, four larval instars, prepupal instar, pupal instar, and adult. Developmental time from egg to adult emergence averaged 29.41 ± 1.85 days, and 47.6% of the eggs developed to adulthood. Female and male survival was 94.42 and 90 days, respectively. Life table parameters show that one female of A. punica is replaced by 86 females (R ₀), the intrinsic growth rate (r _m ) was 0.1115, the average generation time (T) was 40 days, and the doubling time (D _t ) was 6.2 days. The life table parameters suggest that A. punica can be used as a potential predator of C. multicicatrices and, more importantly, provided baseline information for a mass-rearing protocol. This is the first detailed study on the biology of A. punica that reports the potential of this predator as a biological control agent for scale insects of the tribe Iceryini.     

Toxin Gene Contents and Activity of <Emphasis Type="Italic">Bacillus thuringiensis</Emphasis> Strains Against Two Sugarcane Borer Species, <Emphasis Type="Italic">Diatraea saccharalis</Emphasis> (F.) and <Emphasis Type="Italic">D. flavipennella</Emphasis> (Box)

Bacillus thuringiensis (Berliner) bears essential characteristics in the control of insect pests, such as its unique mode of action, which confers specificity and selectivity. This study assessed cry gene contents from Bt strains and their entomotoxicity against Diatraea saccharalis (F.) and Diatraea flavipennella (Box) (Lepidoptera: Crambidae). Bioassays with Bt strains were performed against neonates to evaluate their lethal and sublethal activities and were further analyzed by PCR, using primers to identify toxin genes. For D. saccharalis and D. flavipennella, 16 and 18 strains showed over 30% larval mortality in the 7th day, respectively. The LC₅₀ values of strains for D. saccharalis varied from 0.08 × 10⁵ (LIIT-0105) to 4104 × 10⁵ (LIIT-2707) spores + crystals mL^?1. For D. flavipennella, the LC₅₀ values of strains varied from 0.40 × 10⁵ (LIIT-2707) to 542 × 10⁵ (LIIT-2109) spores + crystals mL^?1. For the LIIT-0105 strain, which was the most toxic to D. saccharalis, the genes cry1Aa, cry1Ab, cry1Ac, cry1B, cry1C, cry1D, cry1F, cry1I, cry2Aa, cry2Ab, cry8, and cry9C were detected, whereas for the strain LIIT-2707, which was the most toxic to D. flavipennella, detected genes were cry1Aa, cry1Ab, cry1Ac, cry1B, cry1D, cry1F, cry1I, cry2Aa, cry2Ab, and cry9. The toxicity data and toxin gene content in these strains of Bt suggest a great variability of activity with potential to be used in the development of novel biopesticides or as source of resistance genes that can be expressed in plants to control pests.     

Toxicological effects of aflatoxin B<Subscript>1</Subscript> on the earthworm <Emphasis Type="Italic">Eisenia fetida</Emphasis> as determined in a contact paper test

In this study, aflatoxin B₁ (AFB₁) toxicity toward the earthworm Eisenia fetida (Savigny 1826) was evaluated in contact paper test systems containing distilled water and ethanol or 20 to 400 μg/ml of AFB₁ over 72 h of exposure. The results indicated that AFB₁ could induce significant damage to earthworms (coiling, curling, excessive mucus secretion, clitellum swelling) at greater than 75 μg/ml. Moreover, AFB₁ had harmful effects on E. fetida (degenerative changes such as bulging of the clitella regions) at levels higher than 150 μg/ml. The calculated LD₅₀ was 168.5 μg/ml. These findings confirm that E. fetida and standardized methods based on this organism (OECD 207 1984) are applicable and useful in mycotoxin related toxicity studies.     

Transmission and characterization of <Emphasis Type="Italic">bla</Emphasis><Subscript>NDM-1</Subscript> in <Emphasis Type="Italic">Enterobacter cloacae</Emphasis> at a teaching hospital in Yunnan,China

Background

In recent years, New Delhi metallo-beta-lactamases 1 (bla _NDM-1) has been reported with increasing frequency and become prevalent. The present study was undertaken to investigate the epidemiological dissemination of the bla _NDM-1 gene in Enterobacter cloacae isolates at a teaching hospital in Yunnan, China.

Methods

Antimicrobial susceptibility testing was performed using VITEK 2 system and E test gradient strips. The presence of integrons and insertion sequence common region 1 were examined by PCR and sequencing. Clonal relatedness was assessed by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing. Conjugation experiments and Southern blot hybridization were performed to determine the transferability of plasmids.

Results

Ten E. cloacae isolates and their Escherichia coli transconjugants were exhibited similar resistant patterns to carbapenems, cephalosporins and penicillins. 8 (80%) of E. cloacae isolates carried class 1 integron and 1 (12.5%) carried class 2 integron. Integron variable regions harbored the genes which encoded resistance to aminoglycosides (aadA1, aadA2, aadA5, aadB, aac(6′)-Ib-cr), sulfamethoxazole/trimethoprim (dfrA17, dfrA12, dfrA15) and Streptozotocin (sat2). Six E. cloacae isolates belonged to ST74 and exhibited highly similar PFGE patterns. Each isolate shared an identical plasmid with ~33.3 kb size that carried the bla _NDM-1 gene, except T3 strain, of which the bla _NDM-1 gene was located on a ~50 kb plasmid.

Conclusions

Our findings suggested that plasmid was able to contribute to the dissemination of bla _NDM-1. Hence, more attention should be devoted to monitor the dissemination of the bla _NDM-1 gene due to its horizontal transfer via plasmid. In addition, nosocomial surveillance system should actively monitor the potential endemic clone of ST74 to prevent their further spread.

     

Decrease of insoluble glucan formation in <Emphasis Type="Italic">Streptococcus mutans</Emphasis> by co-cultivation with <Emphasis Type="Italic">Enterococcus faecium</Emphasis> T7 and glucanase addition

Objectives

To develop preventive canine oral health bio-materials consisting of probiotics and glucanase to reduce insoluble glucan and volatile sulfur compound formation.

Results

Co-cultivation of Enterococcus faecium T7 with Streptococcus mutans at inoculation ratio of 3:1 (v/v) resulted in 25% reduction in the growth of Streptococcus mutans. Amounts of soluble and insoluble glucans produced by S. mutans were decreased to 70 and 55%, respectively. Insoluble glucan was decreased from 0.6 µg/ml in S. mutans culture to 0.03 µg/ml in S. mutans co-cultivated with E. faecium T7 in the presence of Lipomyces starkeyi glucanase. Volatile sulfur compound, a main component of halitosis produced by Fusobacteria nucleatum, was decreased by co-cultivating F. nucleatum with E. faecium.

Conclusion

E. faecium and glucanase can be combined as potentially active ingredients of oral care products for pets by reducing plaque-forming bacteria growth and their by-products that cause cavity and periodontal disease.

     

Testing the cost of reproduction with the rotifer <Emphasis Type="Italic">Brachionus</Emphasis><Emphasis Type="Italic">patulus</Emphasis> at different pH levels

In the present work, we tested the hypothesis that the cost of reproduction was evident under stressful conditions with the rotifer Brachionus patulus at different pH levels (5–10 at 1 unit intervals). We used sublethal pH levels (pH 5, 9, and 10) to simulate stressful conditions. We analyzed the correlations between age-specific fecundity (m ₁, m ₂, m ₃, …) versus future survival (l _x + 1, l _x + 2, l _x + 3, … for the entire lifespan) (survival costs) and future expectation of reproduction (\( V_{ 1}^{*} , \, V_{ 2}^{*} , \, V_{ 3}^{*} , \ldots \) for the entire lifespan) (reproductive costs), using the data obtained from life table demographic studies of B. patulus under stressful and favorable (pH 6, 7, and 8) pH levels. The results showed that significant negative correlations were observed between age-specific fecundity and future survival and future expectation of reproduction at all tested pH levels, indicating that costs of reproduction exist in the rotifer B. patulus under stressful and favorable pH conditions. However, the percentage of statistically significant negative correlations from total correlations of survival and reproductive costs differed greatly, depending on the tested pH conditions. The percentage of significant negative correlation of reproductive costs is significantly higher under stressful pH conditions (pH 5, 9, and 10) than favorable pH conditions (pH 6, 7, and 8). For survival costs, the same trends are also observed, suggesting that the costs of reproduction were more obvious under stressful pH than favorable pH.