A single-amino-acid change in murine norovirus NS1/2 is sufficient for colonic tropism and persistence

Human norovirus (HuNoV) is the major cause of acute nonbacterial gastroenteritis worldwide but has no clear animal reservoir. HuNoV can persist after the resolution of symptoms, and this persistence may be essential for viral maintenance within the population. Many strains of the related murine norovirus (MNV) also persist, providing a tractable animal model for studying norovirus (NoV) persistence. We have used recombinant cDNA clones of representative persistent (CR6) and nonpersistent (CW3) strains to identify a domain within the nonstructural gene NS1/2 that is necessary and sufficient for persistence. Furthermore, we found that a single change of aspartic acid to glutamic acid in CW3 NS1/2 was sufficient for persistence. This same conservative change also caused increased growth of CW3 in the proximal colon, which we found to be a major tissue reservoir of MNV persistence, suggesting that NS1/2 determines viral tropism that is necessary for persistence. These findings represent the first identified function for NoV NS1/2 during infection and establish a novel model system for the study of enteric viral persistence

Weaning stress and gastrointestinal barrier development: Implications for lifelong gut health in pigs

The gastrointestinal (GI) barrier serves a critical role in survival and overall health of animals and humans. Several layers of barrier defense mechanisms are provided by the epithelial, immune and enteric nervous systems. Together they act in concert to control normal gut functions (e.g., digestion, absorption, secretion, immunity, etc.) whereas at the same time provide a barrier from the hostile conditions in the luminal environment. Breakdown of these critical GI functions is a central pathophysiological mechanism in the most serious GI disorders in pigs. This review will focus on the development and functional properties of the GI barrier in pigs and how common early life production stressors, such as weaning, can alter immediate and long-term barrier function and disease susceptibility. Specific stress-related pathophysiological mechanisms responsible for driving GI barrier dysfunction induced by weaning and the implications to animal health and performance will be discussed

Synergistic streptococcal phage λSA2 and B30 endolysins kill streptococci in cow milk and in a mouse model of mastitis

Bovine mastitis results in billion dollar losses annually in the USA alone. Streptococci are among the most relevant causative agents of this disease. Conventional antibiotic therapy is often unsuccessful and contributes to development of antibiotic resistance. Bacteriophage endolysins represent a new class of antimicrobials against these bacteria. In this work, we characterized the endolysins (lysins) of the streptococcal phages λSA2 and B30 and evaluated their potential as anti-mastitis agents. When tested in vitro against live streptococci, both enzymes exhibited near-optimum lytic activities at ionic strengths, pH, and Ca2+ concentrations consistent with cow milk. When tested in combination in a checkerboard assay, the lysins were found to exhibit strong synergy. The λSA2 lysin displayed high activity in milk against Streptococcus dysgalactiae (reduction of CFU/ml by 3.5 log units at 100μg/ml), Streptococcus agalactiae (2 log), and Streptococcus uberis (4 log), whereas the B30 lysin was less effective. In a mouse model of bovine mastitis, both enzymes significantly reduced intramammary concentrations of all three streptococcal species (except for B30 vs. S. dysgalactiae), and the effects on mammary gland wet weights and TNFα concentrations were consistent with these findings. Unexpectedly, the synergistic effect determined for the two enzymes in vitro was not observed in the mouse model. Overall, our results illustrate the potential of endolysins for treatment of Streptococcus-induced bovine mastitis

Synergistic streptococcal phage λSA2 and B30 endolysins kill streptococci in cow milk and in a mouse model of mastitis

Bovine mastitis results in billion dollar losses annually in the USA alone. Streptococci are among the most relevant causative agents of this disease. Conventional antibiotic therapy is often unsuccessful and contributes to development of antibiotic resistance. Bacteriophage endolysins represent a new class of antimicrobials against these bacteria. In this work, we characterized the endolysins (lysins) of the streptococcal phages λSA2 and B30 and evaluated their potential as anti-mastitis agents. When tested in vitro against live streptococci, both enzymes exhibited near-optimum lytic activities at ionic strengths, pH, and Ca2+ concentrations consistent with cow milk. When tested in combination in a checkerboard assay, the lysins were found to exhibit strong synergy. The λSA2 lysin displayed high activity in milk against Streptococcus dysgalactiae (reduction of CFU/ml by 3.5 log units at 100 μg/ml), Streptococcus agalactiae (2 log), and Streptococcus uberis (4 log), whereas the B30 lysin was less effective. In a mouse model of bovine mastitis, both enzymes significantly reduced intramammary concentrations of all three streptococcal species (except for B30 vs. S. dysgalactiae), and the effects on mammary gland wet weights and TNFα concentrations were consistent with these findings. Unexpectedly, the synergistic effect determined for the two enzymes in vitro was not observed in the mouse model. Overall, our results illustrate the potential of endolysins for treatment of Streptococcus-induced bovine mastitis.ISSN:0175-759

Additional file 4: Figure S4. of Sexual dimorphism in the mast cell transcriptome and the pathophysiological responses to immunological and psychological stress

Tnf and Tph1 gene expression in female and male BMMCs. (a) Real-time quantitative PCR of Tnf mRNA transcripts from 6 week-old male and female BMMCs normalized to Rpl4 (n = 3). (b) Real-time quantitative PCR of Tph1 mRNA transcripts from 6 week-old male and female BMMCs normalized to Rpl4 (n = 3). Values represent mean ± SE. †P < 0.10, *P < 0.05 vs. males. (PDF 102 kb

Additional file 3: Figure S3. of Sexual dimorphism in the mast cell transcriptome and the pathophysiological responses to immunological and psychological stress

Sex steroid receptor gene expression in female and male BMMCs. (a–f) Real-time quantitative PCR of Esr1, Esr2, Gper1, Ar, Pgr, Pgrmc2 mRNA transcripts from unstimulated male and female BMMCs normalized to Hrpt (n = 3). Values represent mean ± SE. *P < 0.05, **P < 0.01 vs. males. (PDF 69 kb

Additional file 2: Figure S2. of Sexual dimorphism in the mast cell transcriptome and the pathophysiological responses to immunological and psychological stress

Cytokine mRNA expression of IgE-DNP stimulated female and male BMMCS. (a–f) Real-time quantitative PCR of Tnf, Il6, Il1β, Il4, Il13, and Ccl3 mRNA transcripts from IgE-DNP stimulated male and female BMMCs at 0, 1, 2, and 4 h normalized to Hrpt and relative to 0 h (n = 3). Values represent mean ± SE. *P < 0.05. (PDF 83 kb

Channel-levee evolution in combined contour current–turbidity current flows from flume-tank experiments

Turbidity currents and contour currents are common sedimentary and oceanographic processes in deep-marine settings that affect continental margins worldwide. Their simultaneous interaction can form asymmetric and unidirectionally migrating channels, which can lead to opposite interpretations of paleocontour current direction: channels migrating against the contour current or in the direction of the contour current. In this study, we performed three-dimensional flume-tank experiments of the synchronous interaction between contour currents and turbidity currents to understand the effect of these combined currents on channel architecture and evolution. Our results show that contour currents with a velocity of 10–19 cm s−1 can substantially deflect the direction of turbidity currents with a maximum velocity of 76–96 cm s−1 , and modify the channel-levee system architecture. A lateral and nearly stationary front formed on the levee located upstream of the contour current, reduced overspill and thus restrained the development of a levee on this side of the channel. Sediment was preferentially carried out of the channel at the flank located downstream of the contour current. An increase in contour-current velocity resulted in an increase in channel-levee asymmetry, with the development of a wider levee and more abundant bedforms downstream of the contour current. This asymmetric deposition along the channel suggests that the direction of long-term migration of the channel form should go against the direction of the contour current due to levee growth downstream of the contour current, in agreement with one of the previously proposed conceptual models