Compared with Powdered Lutein, a Lutein Nanoemulsion Increases Plasma and Liver Lutein, Protects against Hepatic Steatosis, and Affects Lipoprotein Metabolism in Guinea Pigs.

BACKGROUND: It is not clear how oil-in-water nanoemulsions of lutein may affect bioavailability and consequently alter lipoprotein metabolism, oxidative stress, and inflammation. OBJECTIVE: The bioavailability as well as effects of a powdered lutein (PL) and an oil-in-water lutein nanoemulsion (NANO; particle size: 254.2 nm; polydispersity index: 0.29; and ζ-potential: -65 mV) on metabolic variables in liver, plasma, and adipose tissue in a guinea pig model of hepatic steatosis were evaluated. METHODS: Twenty-four 2-mo-old male Hartley guinea pigs, weighing 200-300 g (n = 8/group), were fed diets containing 0.25 g cholesterol/100 g to induce liver injury for the duration of the study. They were allocated to control (0 mg lutein), PL (3.5 mg/d), or NANO (3.5 mg/d) groups. After 6 wk, plasma, liver, and adipose tissue were collected for determination of lutein, plasma lipids, tissue cholesterol, and inflammatory cytokines. RESULTS: The NANO group had 2-fold higher concentrations of lutein in plasma (P < 0.001) and 1.6-fold higher concentrations in liver (P < 0.001) than did the PL group, indicating greater bioavailability of this carotenoid. The NANO group also had 24% lower hepatic steatosis scores (P < 0.05), 31% lower hepatic cholesterol accumulation (P < 0.05), and 64% lower plasma alanine aminotransferase (P < 0.05) than did the control group. Hepatic oxidized LDL was 55% lower in both the PL and NANO groups than in the control group (P < 0.05). In plasma, the NANO group had 2-fold higher concentrations of LDL and HDL cholesterol as well as a 2-fold higher number of VLDL, LDL, and HDL particles than did the other 2 groups as evaluated by nuclear magnetic resonance. Furthermore, the NANO group had 15% higher concentrations of free cholesterol in adipose tissue, resulting in higher concentrations of inflammatory markers, than did the other 2 groups. CONCLUSIONS: These results indicate that, although this lutein nanoemulsion exerted protective effects against hepatic steatosis, plasma lipoproteins and adipose tissue cholesterol were increased. These data suggest that the metabolic effects of this particular nanoemulsion might not be protective in all tissues in guinea pigs.

Pathology and diagnosis of necrotic enteritis: is it clear-cut?

The ability to correctly recognize the disease necrotic enteritis (NE) is important not only to those involved in control and treatment of the disease at farm level, but it is also critically important to the search for virulence factors, since a fundamental part of that process is the correct assignation of strains of Clostridium perfringens with respect to virulence. Thus, diagnosticians and investigators need to be able to correctly recognize the lesions of NE. To do this, they must be able to distinguish NE lesions from (1) other enteric diseases such as coccidiosis or viral enteritis, (2) normal features of the intestine, such as the small raised, sometimes red, foci that represent gut-associated lymphoid tissue, (3) autolytic change which may be mistaken for lesions, especially at the microscopical level, by the inexperienced. Errors in diagnosis of NE due to C. perfringens or failure to culture affected areas in which the bacteria of interest with respect to NE are definitively found, might explain some of the early apparently conflicting results with respect to the role of netB in NE. This paper describes at the gross, microscopical and bacteriological level, important features of the intestine of normal poultry and those with NE due to C. perfringens, as well as the common interpretative pitfalls that can lead both to underdiagnosis and overdiagnosis of NE, and to incorrect determination of the virulence of individual C. perfringens strains.

Experimental reproduction of necrotic enteritis in chickens: a review.

This review discusses key factors important in successful experimental reproduction of necrotic enteritis (NE) in chickens, and how these factors can be adjusted to affect the severity of the lesions induced. The critical bacterial factor is the need to use virulent, netB-positive, strains of Clostridium perfringens; disease severity can be enhanced by using netB-positive C. perfringens strains that are also tpeL-positive, by the use of young rather than old broth cultures, and by the number of days of inoculation and the number of bacteria used. Use of cereals rich in non-starch polysaccharides can enhance disease, as does use of animal proteins. Administration of coccidia, including coccidial vaccines, combined with netB-positive C. perfringens, increases the severity of experimentally-induced NE. Dietary manipulation may be less important in coccidia-based models since the latter are so effective. Disease scoring systems and welfare considerations are discussed.

Polyphenol-rich blackcurrant extract exerts hypocholesterolaemic and hypoglycaemic effects in mice fed a diet containing high fat and cholesterol.

Obesity is associated with an increased risk of metabolic abnormalities, such as hyperlipidaemia and hyperglycaemia. We investigated whether polyphenol-rich blackcurrant extract (BCE) can prevent high fat/high cholesterol (HF/HC) diet-induced metabolic disturbances in mice. Male C57BL/6J mice were fed a modified AIN-93M diet containing HF/HC (16% fat, 0·25% cholesterol, w/w) or the same diet supplemented with 0·1% BCE (w/w) for 12 weeks. There were no differences in total body weight and liver weight between groups. Plasma total cholesterol (TC) and glucose levels were significantly lower in BCE group than in controls, while plasma TAG levels were not significantly different. There was a decreasing trend in hepatic TAG levels, and histological evaluation of steatosis grade was markedly lower in the livers of mice fed BCE. Although the mRNA levels of major regulators of hepatic cholesterol metabolism, i.e. 3-hydroxy-3-methyl-glutaryl-CoA reductase (HMGR) and LDL receptor (LDLR), were not significantly altered by BCE supplementation, protein expression of mature sterol-regulatory element-binding protein and LDLR was significantly increased with no change in HMGR protein. The expression of proprotein convertase subtilisin/kexin type 9 that facilitates LDLR protein degradation, as well as one of its transcriptional regulators, i.e. hepatocyte nuclear factor 4α, was significantly decreased in the livers of mice fed BCE. Taken together, BCE supplementation decreased plasma TC and glucose, and inhibited liver steatosis, suggesting that this berry may be consumed to prevent metabolic dysfunctions induced by diets high in fat and cholesterol.

The manganese carbonyl complex [Mn(CO)3(tqa-κ3N)]Br: A novel antimicrobial agent with the potential to treat avian pathogenic Escherichia coli (APEC) infections.

The development of alternatives to antibiotics is essential for the treatment of animal infections and as a measure to reduce the selective pressure on antibiotics that are critical for human medicine. Metal complexes have been highlighted for their antimicrobial activity against several bacterial pathogens. In particular, manganese carbonyl complexes have shown efficacy against multidrug-resistant Gram-negative pathogens, and relatively low cytotoxicity against avian macrophages and in wax moth larval models. They are thus potential candidates for deployment against Avian Pathogenic Escherichia coli (APEC), the aetiological agent of avian colibacillosis, which results in severe animal welfare issues and financial losses worldwide. This study aimed to determine the efficacy of [Mn(CO)3(tqa-κ3N)]Br in Galleria mellonella and chick models of infection against APEC. The results demonstrated in vitro and in vivo antibacterial activity against all antibiotic-resistant APEC test isolates screened in the study.

Toxinotyping of Clostridium perfringens Strains Recovered from U.S. Turkeys with Necrotic Enteritis.

Necrotic enteritis (NE) is a serious disease of chickens and turkeys that causes significant economic losses to the poultry industry. On the basis of studies in chickens, Clostridium perfringens type G is considered by many to be the cause of NE in poultry. However, studies on isolates from Finnish and Italian turkeys with NE revealed that the vast majority were C. perfringens type A, and very few were C. perfringens type G. We therefore examined 74 C. perfringens isolates from U.S. turkeys with NE; 98% were type A and only 1% was type G. This result confirms that different C. perfringens types are involved in NE in turkeys when compared with chickens. We also examined the turkey isolates for other toxin genes associated with enteritis in various animal species, namely tpeL, cpb2, cpe, netE, netF, and netG. The tpeL gene, which has been associated with enhanced virulence of C. perfringens in chickens, was only found in 1% of turkey NE isolates. The cpe gene, which encodes C. perfringens enterotoxin (a major cause of food poisoning and non-foodborne C. perfringens-mediated diarrhea in humans) was also found in only 1% of our turkey NE isolates. Although cpb2, which encodes for the beta2 toxin, was found in 73% of our NE isolates, it has also been found in similar percentages of isolates from turkeys with normal intestine. The netE, netF, and netG genes were not detected among our C. perfringens isolates from turkeys.

Reporte de caso- Tipificación de toxinas de cepas de Clostridium perfringens recuperadas de pavos con enteritis necrótica en los Estados Unidos. La enteritis necrótica es una enfermedad severa de los pollos y pavos que provoca importantes pérdidas económicas a la industria avícola. Sobre la base de estudios en pollos, muchos consideran que Clostridium perfringens tipo G es la causa de la enteritis necrótica en las aves comerciales. Sin embargo, los estudios sobre los aislamientos de pavos con enteritis necrótica de Finlandia e Italia revelaron que la gran mayoría de los aislamientos eran C. perfringens tipo A y muy pocos eran C. perfringens tipo G. Por lo tanto, se examinaron 74 aislamientos de C. perfringens de pavos en los Estados Unidos con enteritis necrótica. El 98% eran del tipo A y solo el 1% fueron del tipo G. Este resultado confirma que diferentes tipos de C. perfringens están involucrados en la enteritis necrótica en pavos en comparación con los pollos. También se examinaron los aislamientos de pavo en busca de otros genes de toxinas asociados con la enteritis en varias especies animales, especialmente, tpeL, cpb2, cpe, netE, netF y netG. El gene tpeL, que se ha asociado con una mayor virulencia de C. perfringens en pollos, solo se encontró en el 1 % de los aislamientos de enteritis necrótica de pavo. El gene cpe, que codifica la enterotoxina de C. perfringens (una de las principales causas de intoxicación alimentaria y diarrea no transmitida por los alimentos causada por C. perfringens en humanos) también se encontró en solo el 1 % de los aislamientos de enteritis necrótica de pavo. Aunque cpb2, que codifica para la toxina beta2, se encontró en el 73 % de los aislamientos de enteritis necrótica, también se ha encontrado en porcentajes similares de aislamientos de pavos con intestino normal. Los genes netE, netF y netG no se detectaron entre los aislamientos de C. perfringens de pavos.

Leiomyosarcoma with widespread metastases in a capybara.

A 10.5-y-old intact female capybara (Hydrochoerus hydrochaeris) with a history of chronic weight loss was euthanized following discovery by palpation of a large intra-abdominal mass. Postmortem examination revealed a large, firm, tan mass expanding the uterine body and extensively adhered to the jejunum and abdominal wall. Numerous pinpoint to 3-cm diameter, tan-to-red, raised masses were present throughout the parietal peritoneum, liver, lungs, and intestinal serosa. Histologic examination of the uterine mass revealed well-differentiated smooth muscle intermixed with abundant collagen, interspersed with a highly anaplastic spindle cell population extending to the serosa; the masses in the lung, liver, and peritoneum were histologically very similar to the anaplastic uterine spindle cells. Immunohistochemical staining of the uterus and lung confirmed smooth muscle origin of the anaplastic cells. To our knowledge, leiomyosarcoma has not been reported previously in a capybara, and the widespread metastases in this case represent an unusually aggressive presentation of this rare malignancy. The animal also had an incidental dermal histiocytoma, a tumor that has also not been reported previously in this species, to our knowledge.

A lutein-enriched diet prevents cholesterol accumulation and decreases oxidized LDL and inflammatory cytokines in the aorta of guinea pigs.

Lutein has been shown to be protective against age-related macular degeneration; however, the antiinflammatory and antioxidant effects of this carotenoid in aortas are less known. Guinea pigs were fed a hypercholesterolemic diet (0.25 g cholesterol/100 g) and randomly allocated to a control group (n = 9) or a lutein group (n = 10) (0.01 g/100 g lutein) [corrected] and fed the experimental diets for 12 wk. Plasma LDL cholesterol and TG did not differ between groups; however, the lutein group had lower concentrations of medium size LDL (P < 0.05). As expected, guinea pigs from the lutein group had higher concentrations of plasma and liver lutein than those from the control group (P < 0.0001). Aortic cholesterol and malondialdehyde concentrations were lower in the lutein group (9.6 ± 2.8 mmol/g and 1.69 ± 1.35 nmol/mg protein) compared to the control group (15.5 ± 2.3 mmol/g and 2.98 ± 1.45 nmol/mg protein) (P < 0.05). Hematoxilin and eosin staining indicated that aortas from the control group presented focal intimal thickening, whereas either less thickness or no visible thickness was present in aortas from the lutein group. Oxidized LDL (oxLDL) was lower both in plasma and aorta in the lutein group compared to the control group (P < 0.001). Aortic cytokines were also lower in the lutein group (P < 0.05). Plasma lutein and oxLDL (r = -0.79; P < 0.0001) and plasma lutein and aortic oxLDL (r = -0.64; P < 0.0001) were negatively correlated. These data suggest that lutein exerts potent antioxidant and antiinflammatory effects in aortic tissue that may protect against development of atherosclerosis in guinea pigs.

Necrotizing pneumonia and pleuritis associated with extraintestinal pathogenic Escherichia coli in a tiger (Panthera tigris) cub.

Extraintestinal pathogenic Escherichia coli (ExPEC) cause diseases in humans and animals, affecting organs outside the alimentary canal. In recent years, ExPEC have been reported as a cause of fatal pneumonia in dogs, cats, and in a horse. In the current report, a fatal case of pneumonia and pleuritis is described in a 4-week-old tiger (Panthera tigris) cub associated with ExPEC. The cub was presented with a sudden-onset respiratory illness and died after a few hours. Postmortem examination of the cub revealed an acute necrotizing pneumonia. The alveolar spaces were filled with large numbers of inflammatory cells (predominantly macrophages), edema, fibrin strands, and short bacillary bacteria. Escherichia coli O6:H31 was isolated in pure culture from the affected lung. It carried virulence genes cnf-1, sfa, fim, hlyD, and papG allele III, which are known to be associated with ExPEC strains. No evidence of infection by any other agent was detected. This is the first report, to the authors' knowledge, in which ExPEC has been associated with pneumonia in tigers.

The ability of disease and non-disease producing strains of Clostridium perfringens from chickens to adhere to extracellular matrix molecules and Caco-2 cells.

Clostridium perfringens is a major enteric pathogen that is responsible for causing necrotic enteritis of poultry. The ability to adhere to the host's intestinal epithelium and to extracellular matrix molecules (ECMM) in the gut, are strategies used by numerous bacterial enteropathogens, however, C. perfringens has received comparatively little attention in this respect. The present study investigated sixteen type A C. perfringens isolates from chickens, with varying disease producing ability with respect to necrotic enteritis in chickens, for their ability to adhere to nine different extracellular matrix molecules (ECMM) and to the intestinal epithelial cell line Caco-2. C. perfringens strains were able to bind to ECMMs and there was strain variation. Strains of C. perfringens that produced severe disease, were capable of binding to collagen type III, IV and V, fibrinogen, laminin and vitronectin at higher levels than less severe disease producing strains, suggesting that the ability to adhere to ECMMs might enhance virulence with respect to induction of necrotic enteritis. In addition, severe disease producing strains also bound better to collagen type III and IV and fibrinogen, than non-disease producing strains. The present study also showed that some strains of C. perfringens possessed the ability to adhere to Caco-2 cells; however no relationship was found between the ability to adhere to Caco-2 cells and disease producing ability.

Cleavable hairpin beacon-enhanced fluorescence detection of nucleic acid isothermal amplification and smartphone-based readout.

Fluorescence detection of nucleic acid isothermal amplification utilizing energy-transfer-tagged oligonucleotide probes provides a highly sensitive and specific method for pathogen detection. However, currently available probes suffer from relatively weak fluorescence signals and are not suitable for simple, affordable smartphone-based detection at the point of care. Here, we present a cleavable hairpin beacon (CHB)-enhanced fluorescence detection for isothermal amplification assay. The CHB probe is a single fluorophore-tagged hairpin oligonucleotide with five continuous ribonucleotides which can be cleaved by the ribonuclease to specifically initiate DNA amplification and generate strong fluorescence signals. By coupling with loop-mediated isothermal amplification (LAMP), the CHB probe could detect Borrelia burgdorferi (B. burgdorferi) recA gene with a sensitivity of 100 copies within 25 min and generated stronger specific fluorescence signals which were easily read and analysed by our programmed smartphone. Also, this CHB-enhanced LAMP (CHB-LAMP) assay was successfully demonstrated to detect B. burgdorferi DNA extracted from tick species, showing comparable results to real-time PCR assay. In addition, our CHB probe was compatible with other isothermal amplifications, such as isothermal multiple-self-matching-initiated amplification (IMSA). Therefore, CHB-enhanced fluorescence detection is anticipated to facilitate the development of simple, sensitive smartphone-based point-of-care pathogen diagnostics in resource-limited settings.

Broad Protection of Pigs against Heterologous PRRSV Strains by a GP5-Mosaic DNA Vaccine Prime/GP5-Mosaic rVaccinia (VACV) Vaccine Boost.

BACKGROUND: Porcine reproductive and respiratory syndrome (PRRS) viruses are a major cause of disease and economic loss in pigs worldwide. High genetic diversity among PRRSV strains is problematic for successful disease control by vaccination. Mosaic DNA and vaccinia (VACV) vaccines were developed in order to improve protection against heterologous PRRSV strains. METHODS: Piglets were primed and boosted with GP5-Mosaic DNA vaccine and recombinant GP5-Mosaic VACV (rGP5-Mosaic VACV), respectively. Pigs vaccinated with rGP5-WT (VR2332) DNA and rGP5-WT VACV, or empty vector DNA and empty VACV respectively, served as controls. Virus challenge was given to separate groups of vaccinated pigs with VR2332 or MN184C. Necropsies were performed 14 days after challenge. RESULTS: Vaccination with the GP5-Mosaic-based vaccines resulted in cellular reactivity and higher levels of neutralizing antibodies to both VR2332 and MN184C PRRSV strains. In contrast, vaccination of animals with the GP5-WT vaccines induced responses only to VR2332. Furthermore, vaccination with the GP5-Mosaic based vaccines resulted in protection against challenge with two heterologous virus strains, as demonstrated by the significantly lower viral loads in serum, tissues, porcine alveolar macrophages (PAMs), and bronchoalveolar lavage (BAL) fluids, and less severe lung lesions after challenge with either MN184C or VR2332, which have only 85% identity. In contrast, significant protection by the GP5-WT based vaccines was only achieved against the VR2332 strain. Conclusions: GP5-Mosaic vaccines, using a DNA-prime/VACV boost regimen, conferred protection in pigs against heterologous viruses.

Prevalence of netB among some clinical isolates of Clostridium perfringens from animals in the United States.

A previously unknown pore forming toxin, called NetB toxin, which is produced by some Australian strains of Clostridium perfringens has recently been reported. This toxin was reported to be critical to the development of the disease necrotic enteritis, in chickens. To investigate the occurrence of the toxin gene (netB) in non-Australian C. perfringens strains, one hundred and six American isolates of C. perfringens were examined. Ninety-two isolates were from chickens, and 14 were from cattle. The netB gene was found in 14 isolates from chickens (7 from chickens with necrotic enteritis, and 7 from unrelated chickens with no evidence of necrotic enteritis). The netB gene was also detected in an isolate recovered from a 3-year-old cow with liver abscesses. The products of all positive netB PCR reactions were sequenced, and these showed 100% nucleotide identity to the netB sequence published in GenBank. Five isolates which had been recovered from five chickens with necrotic enteritis (from four flocks) were netB negative. An additional 24 isolates recovered from one of these lesioned chickens were also netB negative. The present study represents the first study of C. perfringens isolates outside Australia for netB, and the first identification of netB in an isolate from a species other than chickens. The results indicate that the role of NetB in the induction of necrotic enteritis needs to be further investigated, by determining the disease producing capability of both netB positive strains recovered from normal chickens, and netB negative strains recovered from chickens with necrotic enteritis.

Actinomyces hyovaginalis-associated lymphadenitis in a Nubian goat.

A 6-year-old Nubian goat with a history of progressive weight loss and cough was presented for necropsy. The goat tested negative for antibodies to caseous lymphadenitis and caprine arthritis and encephalitis by hemagglutination inhibition assay and enzyme-linked immunosorbent assay, respectively. Postmortem examination revealed marked enlargement and, with histopathology, a fibrinopurulent necrotizing lymphadenitis of a tracheobronchial lymph node, with an appearance similar to that reported in cases of caseous lymphadenitis. An organism characterized by molecular methods as Actinomyces hyovaginalis was isolated together with Staphylococcus spp. and Streptococcus spp. from the lesion. No Corynebacterium pseudotuberculosis was recovered. To the authors' knowledge, this is the first isolation of A. hyovaginalis from a goat. Although the exact contribution of A. hyovaginalis to the lesion remains to be established, this case demonstrates that A. hyovaginalis should be considered in cases of caseous lymphadenitis-type lesions, especially when C. pseudotuberculosis has been excluded.

Canine circoviral hemorrhagic enteritis in a dog in Connecticut.

A 5-mo-old Bassett Hound-Labrador Retriever cross was autopsied following a bout of lethargy, inappetence, and bleeding gums. Mucous membranes were white, and the small intestine was blue-black; the colon contained black feces. The spleen was swollen, and multiple lymph nodes were enlarged and hemorrhagic. Microscopically, the small intestine had focal crypt cell necrosis and circumferential transmural vasculitis, the latter the cause of infarction and the blue-black coloration. Lymphocytes were necrotic in spleen and lymph nodes, and erythrophagocytosis was present in some nodes. Vasculitis was present in brain, meninges, lung, liver, and kidneys. Electron microscopy revealed aggregates of 15-18 nm round viral particles in damaged crypt cells and in the endothelium of small blood vessels. Electron-dense intracytoplasmic inclusions consisting of paracrystalline-arrayed virus were demonstrated in macrophages in medullary lymph node sinuses. These virions were identified as circovirus, which was confirmed by real-time PCR and sequencing.

Liver Glycogen Phosphorylase Deficiency Leads to Profibrogenic Phenotype in a Murine Model of Glycogen Storage Disease Type VI.

Mutations in the liver glycogen phosphorylase (Pygl) gene are associated with the diagnosis of glycogen storage disease type VI (GSD-VI). To understand the pathogenesis of GSD-VI, we generated a mouse model with Pygl deficiency (Pygl -/-). Pygl -/- mice exhibit hepatomegaly, excessive hepatic glycogen accumulation, and low hepatic free glucose along with lower fasting blood glucose levels and elevated blood ketone bodies. Hepatic glycogen accumulation in Pygl -/- mice increases with age. Masson's trichrome and picrosirius red staining revealed minimal to mild collagen deposition in periportal, subcapsular, and/or perisinusoidal areas in the livers of old Pygl -/- mice (>40 weeks). Consistently, immunohistochemical analysis showed the number of cells positive for alpha smooth muscle actin (α-SMA), a marker of activated hepatic stellate cells, was increased in the livers of old Pygl -/- mice compared with those of age-matched wild-type (WT) mice. Furthermore, old Pygl -/- mice had inflammatory infiltrates associated with hepatic vessels in their livers along with up-regulated hepatic messenger RNA levels of C-C chemokine ligand 5 (Ccl5/Rantes) and monocyte chemoattractant protein 1 (Mcp-1), indicating inflammation, while age-matched WT mice did not. Serum levels of aspartate aminotransferase and alanine aminotransferase were elevated in old Pygl -/- mice, indicating liver damage. Conclusion: Pygl deficiency results in progressive accumulation of hepatic glycogen with age and liver damage, inflammation, and collagen deposition, which can increase the risk of liver fibrosis. Collectively, the Pygl-deficient mouse recapitulates clinical features in patients with GSD-VI and provides a model to elucidate the mechanisms underlying hepatic complications associated with defective glycogen metabolism.

Dietary Prevention of Colitis by Aronia Berry is Mediated Through Increased Th17 and Treg.

SCOPE: Increased fruit consumption is associated with reduced risk of colitis. It has been investigated whether the anti-colitic effects of the polyphenol-rich aronia berry (Aronia mitschurinii 'Viking') are mediated through Th17 and Treg. METHODS AND RESULTS: Colitis is induced in recombinase activating gene-1 deficient mice injected with syngeneic CD4+ CD62L+ naïve T cells. Mice consume either 4.5% w/w aronia-berry-supplemented or a control diet concurrent with T cell transfer. The extent of colitis and immunocyte populations are evaluated at weeks 3 to 7 after transfer. Aronia consumption prevents colitic wasting and reduces colon weight/length ratios relative to the control diet at weeks 5 and 7. Compared to the control diet, aronia feeding increases Treg in mesenteric lymph node at all colitis stages. Treg and regulatory Th17 subpopulations (IL-17A+ IL-10+ and IL-17A+ IL-22+ ) are increased in lamina propria and spleen at week 5 in aronia-fed mice. Aronia feeding also decreases total CD4+ cells but increases colonic Tregs. The ability of aronia to modulate colonic cytokines is associated with functional T cell IL-10 and increased diversity of microbiota. CONCLUSIONS: Aronia berry consumption inhibits adoptive transfer colitis by increasing Treg and regulatory Th17 cells. Dietary modulation of T cells is dynamic and precedes colitic wasting.

A PRRSV GP5-Mosaic vaccine: Protection of pigs from challenge and ex vivo detection of IFNγ responses against several genotype 2 strains.

Porcine reproductive and respiratory syndrome virus (PRRSV), is a highly mutable RNA virus that affects swine worldwide and its control is very challenging due to its formidable heterogeneity in the field. In the present study, DNA vaccines constructed with PRRSV GP5-Mosaic sequences were complexed to cationic liposomes and administered to experimental pigs by intradermal and intramuscular injection, followed by three boosters 14, 28 and 42 days later. The GP5-Mosaic vaccine thus formulated was immunogenic and induced protection from challenge in vaccinated pigs comparable to that induced by a wild type (VR2332) GP5 DNA vaccine (GP5-WT). Periodic sampling of blood and testing of vaccine-induced responses followed. Interferon-γ (IFN-γ) mRNA expression by virus-stimulated peripheral blood mononuclear cells (PBMCs) of GP5-Mosaic-vaccinated pigs was significantly higher compared to pigs vaccinated with either GP5-WT or empty vector at 21, 35 and 48 days after vaccination. Cross-reactive cellular responses were also demonstrated in GP5-Mosaic vaccinated pigs after stimulation of PBMCs with divergent strains of PRRSV. Thus, significantly higher levels of IFN-γ mRNA were detected when PBMCs from GP5-Mosaic-vaccinated pigs were stimulated by four Genotype 2 strains (VR2332, NADC9, NADC30 and SDSU73), which have at least 10% difference in GP5 amino acid sequences, while such responses were recorded only upon VR2332 stimulation in GP5-WT-vaccinated pigs. In addition, the levels of virus-specific neutralizing antibodies were higher in GP5-Mosaic or GP5-WT vaccinated pigs than those in vector-control pigs. The experimental pigs vaccinated with either the GP5-Mosaic vaccine or the GP5-WT vaccine were partially protected from challenge with VR2332, as measured by significantly lower viral loads in sera and tissues and lower lung lesion scores than the vector control group. These data demonstrate that the GP5-Mosaic vaccine can induce cross-reactive cellular responses to diverse strains, neutralizing antibodies, and protection in pigs.

Correction: A PRRSV GP5-Mosaic vaccine: Protection of pigs from challenge and ex vivo detection of IFNγ responses against several genotype 2 strains.

[This corrects the article DOI: 10.1371/journal.pone.0208801.].

Green tea extract protects leptin-deficient, spontaneously obese mice from hepatic steatosis and injury.

The incidence of nonalcoholic fatty liver disease (NAFLD) has risen along with the ongoing obesity epidemic. Green tea extract (GTE) inhibits intestinal lipid absorption and may regulate hepatic lipid accumulation. The objective of this study was to determine whether GTE protects against hepatic lipid accumulation during the development of NAFLD in an obese mouse model. Five-wk-old ob/ob (obese) mice and their lean littermates (8 mice x genotype(-1) x dietary treatment(-1)) were fed GTE at 0, 1, or 2% (wt:wt) for 6 wk. The body weights of obese mice and lean littermates fed diets containing GTE were 23-25% and 11-20% lower (P < 0.05) than their respective controls fed no GTE. Histologic evaluation showed a significant reduction in hepatic steatosis in GTE-fed obese mice only and histologic scores were correlated with hepatic lipid concentration (r = 0.84; P < 0.05), which was reduced dose dependently by GTE. GTE protected against hepatic injury as suggested by 30-41% and 22-33% lower serum alanine aminotransferase and aspartate aminotransferase activities, respectively. Hepatic alpha-tocopherol was 36% higher in obese mice than lean mice. GTE tended (P = 0.06) to lower hepatic alpha-tocopherol, which was not fully explained by the GTE-mediated reduction in hepatic lipid. Hepatic ascorbic acid was lower in obese mice than in lean mice (P < 0.05) and was unaltered by GTE. Obese mice had lower serum adiponectin than lean mice and this was not affected by GTE. The results suggest that GTE protects against NAFLD by limiting hepatic lipid accumulation and injury without affecting hepatic antioxidant status and adiponectin-mediated lipid metabolism. Further study is underway to define the events by which GTE protects against obesity-triggered NAFLD.