Follicle-stimulating hormone increases the intramuscular fat content and expression of lipid biosynthesis genes in chicken breast muscle

Intramuscular fat (IMF) is a crucial factor in the quality of chicken meat. The genetic basis underlying it is complex. Follicle-stimulating hormone (FSH), well-known as an effector in reproductive tissues, was recently discovered to stimulate abdominal fat accumulation in chicken. The effect of FSH on IMF accumulation and the underlying molecular regulatory mechanisms controlling both IMF and abdominal fat deposition in vivo are largely unknown. In this study, two groups of chickens were treated with chicken FSH or a placebo. The lipid content of breast muscle, abdominal fat volume, and serum concentrations of FSH were examined. Related genes implicated in breast muscle and abdominal fat accumulation were also investigated. Compared to the control group, the triglyceride (TG) content of breast muscle and the percentage of abdominal fat in FSH-treated chickens were significantly increased by 64.9% and 56.5% (P<0.01), respectively. The FSH content in the serum of FSH-treated chickens was 2.1 times than that of control chickens (P<0.01). Results from quantitative real-time polymerase chain reaction (qRT-PCR) assays showed that relative expression levels of fatty acid synthase (FAS), lipoprotein lipase (LPL), diacylglycerol acyltransferase 2 (DGAT2), adipocyte fatty acid binding protein (A-FABP), and peroxisome proliferator-activated receptor γ (PPARγ) were significantly upregulated in breast muscle following FSH treatment (P<0.01). Treatment with FSH also significantly increased relative expression levels of FAS, LPL, DGAT2, A-FABP, and PPARγ in abdominal fat tissue (P<0.05). The results of principal component analysis (PCA) for gene expression (breast muscle and abdominal fat) showed that the control and FSH treatment groups were well separated, which indicated the reliability of the data. This study demonstrates that FSH plays an important role in IMF accumulation in female chickens, which likely involves the regulation of biosynthesis genes related to lipid metabolism.     

<Emphasis Type="Italic">Bildung</Emphasis> in globalizing times

This key paper is intended to produce a comparison of Bildung and globalization, acknowledging that Bildung and globalization are both fuzzy concepts. Our comparison of globalization and Bildung therefore is to be seen as a construction. To reach this objective, we start with the identification of dimensions of the concept of globalization based predominantly on sociological research. This analysis is followed by explication of dimensions of the concept of Bildung as they can be identified before a frame of globalization. We want to find out whether it makes sense to see globalization as one part of a new model of Bildung. In the conclusion, we demonstrate the normativity of each and any theory of Bildung in globalizing times, and we explain why it is necessary to reflect on the normativity of the different concepts. We indicate where we see connections with the other papers of this special issue of “Zeitschrift für Erziehungswissenschaft”.     

<Emphasis Type="Italic">Pediococcus Acidilactici</Emphasis> Inhibit Biofilm Formation of Food-Borne Pathogens on Abiotic Surfaces

In this study, we aimed to examine the inhibitory effect of PA003, a Pediococcus acidilactici that produces lactic acid and antimicrobial peptides pediocin, on pathogenic biofilm formation on abiotic surfaces. PA003 and pathogens (Escherichia coli, Salmonella enterica serovar Typhimurium, Staphylococcus aureus and Listeria monocytogenes) were used to evaluate auto-aggregation, hydrophobicity, biofilm formation and biofilm formation inhibition on stainless steel, polyvinyl chloride and glass slides in terms of exclusion, displacement and competition. The results showed the highest auto-aggregation abilities were observed for one of the E. coli strains EAggEC (E58595) and the highest hydrophobic strain was observed with EPEC (E2348/69) (51.9%). The numbers of biofilm cells of E. coli, S. Typhimurium, S. aureus and L. monocytogenes on stainless steel, polyvinyl chloride and glass slide coupons were effectively reduced by approximately 4 log CFU/coupon. These results demonstrate that lactic acid bacteria can be used as an alternative to effectively control the formation of biofilms by food-borne pathogens.     

Polymorphisms of the <Emphasis Type="Italic">IGF1</Emphasis> gene and their association with growth traits,serum concentration and expression rate of <Emphasis Type="Italic">IGF1</Emphasis> and <Emphasis Type="Italic">IGF1R</Emphasis> in buffalo

The insulin-like growth factor 1 (IGF1) gene is a member of the group of somatotropin axis genes that play a significant role in cell proliferation and growth of muscles. Here, we searched for polymorphisms in buffalo IGF1 and found two novel single nucleotide polymorphisms (SNPs), G64A and G280A, in the noncoding sequences of exon 1 and exon 4, respectively. Statistical analysis of different genotypes showed that the individuals with GG genotypes had significantly (P<0.05) higher body weight (BW) and average daily gain (ADG) than those with other genotypes at ages of 3–6 months in G64A SNP and 6–9 months in G280A SNP. The combined genotypes of these two SNPs produced three haplotypes, GG/GG, AG/AG, and AA/AA, which were significantly associated (P<0.0001) with BW and ADG at an age from 3 to 12 months. Buffaloes with the homozygous GG/GG haplotype showed higher growth performance than other buffaloes. The two SNPs were correlated with mRNA levels of IGF1 and IGF1 receptor (IGF1R) in semitendinosus muscle as well as with the serum concentration level of IGF1. Also, buffaloes with GG/GG haplotype showed higher mRNA and serum concentration levels. The data revealed that these two SNPs could be valuable genetic markers for selection of Egyptian buffaloes for better performance in the population.     

Green Fluorescent Protein Recombinant Nisin as a Probe for Detection of Gram-Positive Bacteria

A great amount of foodborne pathogens were Gram-positive(G+) bacteria, a threat to public health. In this study, considering the binding ability of nisin towards G+ bacteria and the stable fluorescent ability of EGFP protein, a fluorescent nisin–EGFP protein probe was constructed by a gene engineering method. Nisin and EGFP were used as the receptor and fluorophore, respectively, to detect G+ bacteria. The nisin and egfp gene were amplified separately according to the sequence published in Gen Bank using unique primers. The two genes were cloned into a pET-28b(+) vector resulting in apET-28b(+)–nisin–egfp vector. The vector was transferred into Escherichia coli(E. coli) BL21(DE3) for expression. The expressed protein was extracted, purified by a Ni–NTA column, and then tested by the SDS-PAGE method to confirm its molecular weight. Listeria monocytogenes(L.monocytogenes), Staphylococcus aureus(S. aureus), and Micrococcus luteus(M. luteus) were used as the representations of G+ bacteria. E. coli O157, representing the gram-negative(G-) bacteria, was used as a negative control. The binding specificity of the recombinant protein was performed on two types of bacteria and then detected through fluorescent microscopy. The results indicated that the nisin–EGFP probe could detect G+ bacteria at 10~8CFU/mL.     

Effects of rhizobacteria <Emphasis Type="Italic">Paenibacillus polymyxa</Emphasis> APEC136 and <Emphasis Type="Italic">Bacillus subtilis</Emphasis> APEC170 on biocontrol of postharvest pathogens of apple fruits

In this study, plant growth-promoting rhizobacteria (PGPR) were evaluated as potential biocontrol agents against postharvest pathogens of apple fruits. In vitro bioassays revealed that, out of 30 isolates screened, isolates APEC136 and APEC170 had the most significant inhibitory effects against the mycelial growth of several fungal pathogens. Analysis of 16S ribosomal RNA (rRNA) sequences identified the two effective isolates as Paenibacillus polymyxa and Bacillus subtilis, respectively. The two strains showed greater growth in brain-heart infusion broth than in other growth media. Treatment of harvested apples with suspensions of either strain reduced the symptoms of anthracnose disease caused by two fungal pathogens, Colletotrichum gloeosporioides and Colletotrichum acutatum, and white rot disease caused by Botryosphaeria dothidea. Increased productions of amylase and protease by APEC136, and increased productions of chitinase, amylase, and protease by APEC170 might have been responsible for inhibiting mycelial growth. The isolates caused a greater reduction in the growth of white rot than of anthracnose. These results indicate that the isolates APEC136 and APEC170 are promising agents for the biocontrol of anthracnose and white rot diseases in apples after harvest, and suggest that these isolates may be useful in controlling these diseases under field conditions.     

Glycyrrhizic acid activates chicken macrophages and enhances their <Emphasis Type="Italic">Salmonella</Emphasis>-killing capacity in vitro

Objective

Salmonella enterica remains a major cause of food-borne disease in humans, and Salmonella Typhimurium (ST) contamination of poultry products is a worldwide problem. Since macrophages play an essential role in controlling Salmonella infection, the aim of this study was to evaluate the effect of glycyrrhizic acid (GA) on immune function of chicken HD11 macrophages.

Methods

Chicken HD11 macrophages were treated with GA (0, 12.5, 25, 50, 100, 200, 400, or 800 μg/ml) and lipopolysaccharide (LPS, 500 ng/ml) for 3, 6, 12, 24, or 48 h. Evaluated responses included phagocytosis, bacteria-killing, gene expression of cell surface molecules (cluster of differentiation 40 (CD40), CD80, CD83, and CD197) and antimicrobial effectors (inducible nitric oxide synthase (iNOS), NADPH oxidase-1 (NOX-1), interferon-γ (IFN-γ), LPS-induced tumor necrosis factor (TNF)-α factor (LITAF), interleukin-6 (IL-6), and IL-10), and production of nitric oxide (NO) and hydrogen peroxide (H₂O₂).

Results

GA increased the internalization of both fluorescein isothiocyanate (FITC)-dextran and ST by HD11 cells and markedly decreased the intracellular survival of ST. We found that the messenger RNA (mRNA) expression of cell surface molecules (CD40, CD80, CD83, and CD197) and cytokines (IFN-γ, IL-6, and IL-10) of HD11 cells was up-regulated following GA exposure. The expression of iNOS and NOX-1 was induced by GA and thereby the productions of NO and H₂O₂ in HD11 cells were enhanced. Notably, it was verified that nuclear factor-κB (NF-κB) and c-Jun N-terminal kinase (JNK) pathways were responsible for GA-induced synthesis of NO and IFN-γ gene expression.

Conclusions

Taken together, these results suggested that GA exhibits a potent immune regulatory effect to activate chicken macrophages and enhances Salmonella-killing capacity.

     

Fungal spondylodiscitis in a patient recovered from H7N9 virus infection: a case study and a literature review of the differences between <Emphasis Type="Italic">Candida</Emphasis> and <Emphasis Type="Italic">Aspergillus</Emphasis> spondylodiscitis

To report a rare case of fungal spondylodiscitis in a patient recovered from H7N9 virus infection and perform a literature review of the different characteristics of Candida and Aspergillus spondylodiscitis, we reviewed cases of spondylodiscitis caused by Candida and Aspergillus species. Data, including patients’ information, pathogenic species, treatment strategy, outcomes, and relapses, were collected and summarized. The characteristics of Candida and Aspergillus spondylodiscitis were compared to see if any differences in clinical features, management, or consequences could be detected. The subject of the case study was first misdiagnosed as having a vertebral tumor, and then, following open biopsy, was diagnosed as having fungal spondylodiscitis. The patient made a good recovery following radical debridement. Seventy-seven additional cases of Candida spondylodiscitis and 94 cases of Aspergillus spondylodiscitis were identified in the literature. Patients with Candida spondylodiscitis tended to have a better outcome than patients with Aspergillus spondylodiscitis (cure rate 92.3% vs. 70.2%). Candida was found more frequently (47.8%) than Aspergillus (26.7%) in blood cultures, while neurological deficits were observed more often in patients with Aspergillus spondylodiscitis (43.6% vs. 25.6%). Candida spinal infections were more often treated by radical debridement (60.5% vs. 39.6%). Patients with Candida spondylodiscitis have better outcomes, which may be associated with prompt recognition, radical surgical debridement, and azoles therapy. A good outcome can be expected in fungal spondylodiscitis with appropriate operations and anti-fungal drugs.     

An oriental melon 9-lipoxygenase gene <Emphasis Type="Italic">CmLOX09</Emphasis> response to stresses,hormones, and signal substances

In plants, lipoxygenases (LOXs) play a crucial role in biotic and abiotic stresses. In our previous study, five 13-LOX genes of oriental melon were regulated by abiotic stress but it is unclear whether the 9-LOX is involved in biotic and abiotic stresses. The promoter analysis revealed that CmLOX09 (type of 9-LOX) has hormone elements, signal substances, and stress elements. We analyzed the expression of CmLOX09 and its downstream genes—CmHPL and CmAOS—in the leaves of four-leaf stage seedlings of the oriental melon cultivar “Yumeiren” under wound, hormone, and signal substances. CmLOX09, CmHPL, and CmAOS were all induced by wounding. CmLOX09 was induced by auxin (indole acetic acid, IAA) and gibberellins (GA₃); however, CmHPL and CmAOS showed differential responses to IAA and GA₃. CmLOX09, CmHPL, and CmAOS were all induced by hydrogen peroxide (H₂O₂) and methyl jasmonate (MeJA), while being inhibited by abscisic acid (ABA) and salicylic acid (SA). CmLOX09, CmHPL, and CmAOS were all induced by the powdery mildew pathogen Podosphaera xanthii. The content of 2-hexynol and 2-hexenal in leaves after MeJA treatment was significantly higher than that in the control. After infection with P. xanthii, the diseased leaves of the oriental melon were divided into four levels—levels 1, 2, 3, and 4. The content of jasmonic acid (JA) in the leaves of levels 1 and 3 was significantly higher than that in the level 0 leaves. In summary, the results suggested that CmLOX09 might play a positive role in the response to MeJA through the hydroperoxide lyase (HPL) pathway to produce C6 alcohols and aldehydes, and in the response to P. xanthii through the allene oxide synthase (AOS) pathway to form JA.     

Improving the antioxidant activity and enriching salvianolic acids by the fermentation of <Emphasis Type="Italic">Salvia miltiorrhizae</Emphasis> with <Emphasis Type="Italic">Geomyces luteus</Emphasis>

The antioxidant activities and total phenolic content of fermented Salvia miltiorrhiza with fungus Geomyces luteus were investigated. The results revealed that G. luteus fermentation could significantly improve the antioxidant activity and total phenolic content of S. miltiorrhiza. The main antioxidant constituents were characterized by spectroscopic analysis as salvianolic acids. High-performance liquid chromatography (HPLC) quantification also showed the enhanced content of salvianolic acid B after fermentation. The present study suggests that G. luteus fermentations are effective in the S. miltiorrhiza salvianolic acids’ enrichment process.     

Endophytes from medicinal plants and their potential for producing indole acetic acid,improving seed germination and mitigating oxidative stress

Medicinal plants have been used by marginal communities to treat various ailments. However, the potential of endophytes within these bio-prospective medicinal plants remains unknown. The present study elucidates the endophytic diversity of medicinal plants (Caralluma acutangula, Rhazya stricta, and Moringa peregrina) and the endophyte role in seed growth and oxidative stress. Various organs of medicinal plants yielded ten endophytes, which were identified as Phoma sp. (6 isolates), Alternaria sp. (2), Bipolaris sp. (1), and Cladosporium sp. (1) based on 18S rDNA sequencing and phylogenetic analysis. The culture filtrates (CFs; 25%, 50%, and 100% concentrations) from these endophytes were tested against the growth of normal and dwarf mutant rice lines. Endophytic CF exhibited dose-dependent growth stimulation and suppression effects. CF (100%) of Phoma sp. significantly increased rice seed germination and growth compared to controls and other endophytes. This growth-promoting effect was due to the presence of indole acetic acid in endophytic CF. The gas chromatography/mass spectrometry (GC/MS) analysis showed the highest indole acetic acid content ((54.31±0.21) μmol/L) in Bipolaris sp. In addition, the isolate of Bipolaris sp. exhibited significantly higher radical scavenging and anti-lipid peroxidation activity than the other isolates. Bipolaris sp. and Phoma sp. also exhibited significantly higher flavonoid and phenolic contents. The medicinal plants exhibited the presence of bio-prospective endophytic strains, which could be used for the improvement of crop growth and the mitigation of oxidative stresses.     

Vector refinement equation and subdivision schemes in <Emphasis Type="Italic">L</Emphasis><Subscript>p</Subscript> spaces

In this paper we will first prove that the nontrivial L _p solutions of the vector refinement e-quation exist if and only if the corresponding subdivision scheme with a suitable initial function converges in L _p without assumption of the stability of the solutions. Then we obtain a characterization of the convergence of the subdivision scheme in terms of the mask. This gives a complete answer to the existence of L _p solutions of the refinement equation and the convergence of the corresponding subdivision schemes.     

Alteration of <Emphasis Type="Italic">SlYABBY2b</Emphasis> gene expression impairs tomato ovary locule number and endogenous gibberellin content

Tomato is an ideal model species for fleshy fruit development research. SlYABBY2b regulates the ovary locule number, which is increased by gibberellins, in tomato. However, the relationship between SlYABBY2b and endogenous gibberellin is poorly understood. In this study, SlYABBY2b-overexpressing and RNA interference (RNAi) transgenic tomato plants were used to elucidate the mechanism by which SlYABBY2b regulates the ovary locule number and endogenous gibberellin content in tomato. SlYABBY2b-overexpressing plants showed fewer locules and lower gibberellin content than the control plants. Contrasting results were found in the RNAi lines. Therefore, the SlYABBY2b gene negatively regulates tomato ovary locule number and endogenous gibberellin content. Furthermore, the expression of SlYABBY2b gene was remarkably higher than that of the wild type in the apical shoots of gibberellindeficient mutants. This showed that the gibberellins can inhibit the expression of SlYABBY2b gene negative regulation. Further study revealed that SlYABBY2b suppressed the expression of SlGA20ox1 and SlGA3ox2, but increased that of SlGA2ox1 and SlGA2ox5 in the apical shoots of SlYABBY2b-overexpressing plants, thereby reducing gibberellin content. Contrasting results were found in the RNAi lines. Our results showed that the SlYABBY2b gene was located on gibberellin signal transduction pathways, fed back regulation of the synthesis of gibberellin, and felt exogenous gibberellin signal to further regulate the formation of tomato locule.     

Cloning and efficient expression of <Emphasis Type="Italic">Bacillus sp.</Emphasis> BH072 <Emphasis Type="Italic">tasA</Emphasis> gene in <Emphasis Type="Italic">Escherichia coli</Emphasis>

The Bacillus strain BH072 isolated from a honey sample showed strong antifungal activity against phytopathogen. Gene cloning test demonstrated that the strain had a tasA gene encoding an antifungal TasA protein. Although the wild strain simultaneously produced various antifungal substances, only the physicochemical property and antifungal activity of TasA protein were unclear due to the difficulty in extraction. In this study, tasA gene encoding the protein from Bacillus sp. BH072 was amplified by using the polymerase chain reaction (PCR) method and cloned into pET 28a (+) vector, and then expressed in host cells Escherichia coli BL21 (DE3). The expressed proteins were collected by centrifugation and ultrasonic treatment, and then purified by using nickel-nitrilotriacetic acid (Ni-NTA) metal affinity column and dialysis methods. The result of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) test showed that an expected protein band appeared with a size of 31 kDa. The expressed products possessed antifungal activity against the phytopathogenic indicator strain Botrytis cinerea. A genetically engineered strain tasA of E. coli was established in this study which can efficiently express Tas A protein.     

Effects of temperature and diet on length-weight relationship and condition factor of the juvenile Malabar blood snapper (<Emphasis Type="Italic">Lutjanus malabaricus</Emphasis> Bloch &amp; Schneider, 1801)

In this study we aimed to analyze the effects of water temperature and diet on the length-weight relationship and condition of juvenile Malabar blood snapper Lutjanus malabaricus over a 30-d experimental period. The experiment was conducted in the laboratory using a flow-through-sea-water system. The fish were subjected to four different temperatures (22, 26, 30, and 34 °C) and two diets (commercial pellet and natural shrimp). Fish were fed twice daily. L. malabaricus exhibited negative allometric growth (b<3) at the beginning of the experiment (Day 0) at all temperatures and both diets except for 22 °C fed with shrimp, which showed isometric growth (b=3). Conversely, at the end of the experiment (Day 30) fish showed isometric growth (b=3) at 30 °C fed with the pellet diet, indicating that the shape of the fish did not change with increasing weight and length, and a positive allometric growth (b>3) at 30 °C fed with shrimp diet, which indicated that fish weight increases faster than their length. The rest of the temperatures represented negative allometric growth (b<3) on both diet, meaning that fish became lighter with increasing size. The condition factors in the initial and final measurements were greater than 1, indicating the state of health of the fish, except for those fed on a pellet diet at 34 °C. However, the best condition was obtained at 30 °C on both diets. Nevertheless, diets did not have a significant effect on growth and condition of juvenile L. malabaricus. The data obtained from this study suggested culturing L. malabaricus at 30 °C and feeding on the pellet or shrimp diet, which will optimize the overall production and condition of this commercially important fish species.     

Prevalence and genotype of <Emphasis Type="Italic">Chlamydia psittaci</Emphasis> in faecal samples of birds from zoos and pet markets in Kunming,Yunnan, China

Chlamydia psittaci is an important zoonotic pathogen in birds and may be transmitted to humans and result in severe respiratory disease. To assess the prevalence and genotype of C. psittaci in birds in Kunming, Yunnan, China, a total of 136 specimens of psittacine birds involving 8 species were collected from the city’s zoos (n=60) and pet markets (n=76). The frequency of C. psittaci infection was 19.9% (27/136) in the psittacine birds. The prevalence of C. psittaci was higher in pet birds (26.3%; 20/76) than in zoo birds (11.7%; 7/60) (P=0.034). In particular, among Agapornis fischeri, the C. psittaci infection (50%; 10/20) was significantly more frequent in the pet markets than in the zoos (P=0.006). In addition, the highest prevalence of 41.2% (7/17) was found in Ara ararauna. To determine the genotype of C. psittaci, 23 OmpA gene fragments (about 1.4 kb) in 27 positive samples were successfully amplified and sequenced. Phylogenetic analysis showed that all the 23 strains belonged to genotype A. Our results demonstrate the high prevalence of C. psittaci genotype A infection in psittacine birds in Yunnan Province, suggesting a potential threat to human health in this area. Therefore, it is necessary to take effective measures to prevent the spread of C. psittaci among psittacine birds, as well as among employees and customers.