Foraminiferal Evidence of Late Holocene Sea-Level Change and Amerindian Site Distribution at Montague Harbour, British Columbia

Foraminiferal and sedimentological analysis of an underwater stratigraphie section from an Amerindian habitation site at Montague Harbour, British Columbia has further documented late Holocene sea level changes. It appears that part of the documented transgression was caused by tectonic subsidence of the area (Event 1 at approx. 3500 calendar years BP and Event 2 sometime before 1100 calendar years BP) and was recognized in the stratigraphie record by rapid environmental changes. The environmental changes caused by rapid shifts in water depth were recognized through sedimentological and foraminiferal evidence. The tectonic subsidence events, coupled with gentle late Holocene transgression, caused the breaching of Montague Harbour's northwestern channel. The breaching of the channel improved water circulation and increased salinity within the harbour. The salinity changes are reflected in the shift from a low salinity Cribroelphidium excavatum (Terquem, 1876) phenotype "clavata" dominated biofacies (1) at the base of the section to a higher salinity Buccella tenerrima (Bandy, 1950) and Elphidiella hannai (Cushman and Grant, 1927) dominated biofacies (2) at the top. These sea-level changes would have eventually forced local Amerindian settlements inland. The 14C dating of wood and shell, indicates that the recovery of archaeological remains of the Charles culture (ca.6500-3200 years BP) requires investigation in deeper waters.L'analyse des foraminifères et des sédiments d'une coupe stratigraphique sous-marine d'un habitat amérindien a permis d'accroître les connaissances sur les changements du niveau marin. Il semble qu'une partie de la transgression connue ait été causée par une subsidence de nature tectonique (1er épisode vers 3500 cal. BP et 2e épisode un peu avant 1100 cal. BP) qui a entraîné des changements environnementaux rapides consignés dans la stratigraphie. Les sédiments et les foraminifères ont en effet enregistré les changements rapides de la profondeur de l'eau. Les épisodes de subsidence, accompagnés par une transgression modérée à l'Holocène supérieur, ont provoqué la formation d'une brèche dans le chenal nord-ouest du Montague Harbour, qui a facilité la circulation des eaux et fait augmenter la salinité à l'intérieur du havre. Ce changement de salinité s'est manifesté par le remplacement du biofaciès (1) de faible salinité dominé par Cribroelphidium excavatum (Terquem, 1876) type « clavata », à la base de la coupe, par le biofaciès (2) de forte salinité dominé par Bucella tenerrima (Bandy, 1950) et Elphidiella hannai (Cushman et Grant, 1927) au sommet. Les changements du niveau marin ont forcé les Amérindiens à déménager vers l'intérieur. La datation au 14C sur bois et coquille montre que la récupération de vestiges archéologiques de la culture Charles (ca 6500-3200 BP) devra se faire en eaux plus profondes.Die Foraminiferen- und Sediment-Analyse eines stratigraphischen Unterwasserabschnitts von einer Indianer-Siedlung am Montague Harbour, British Columbia, hat weitere Belege ùber Meeresniveauwechsel im spàten Holozàn geliefert. Es scheint, daB ein Teil der dokumentierten Transgression durch tekton-ische Senkung des Gebiets verursacht wurde (Ereignis 1 urn circa 3500 Kalenderjahre v.u.Z. und Ereignis 2 irgendwann vor 1100 Kalenderjahren v.u.Z.) und in dem stratigraphischen Beleg durch rasche Umweltverànderungen kenntlich wurde. Die durch schnelle Wechsel der Wassertiefe verursachten Umweltverànderungen sind durch Sediment- und Foraminiferen-Belege aufgezeichnet worden. Die Episoden tektonischer Senkung verursachten zusammen mit einer sanften Transgression im spàten Holozàn die Bildung einer Bresche im nordwestlichen Kanal von Montague Harbour. Die Bresche im Kanal verbesserte die Wasserzirkulation und erhôhte den SaIzgehalt im Hafen. Die Verànderungen im Salzgehalt spiegeln sich im Wechsel von einer Biofazies (1) mit einem niedrigen Salzgehalt beherrscht von Cribroelphidium excavatum (Terquem, 1876) des Phânotypus "clavata" an der Basis des Schnittes, zu einer Biofazies (2) mit hôherem Salzgehalt, beherrscht von Bucella tenerrima (Bandy, 1950) und Elphidiella hannai (Cushman und Grant, 1927) an der Spitze. Diese Meeresniveau-Wechsel haben wohl die ôrtlichen indianischen Siedlungen gezwungen, landeinwàrts zu Ziehen. Die '4C-Datierung von HoIz und Muscheln zeigt, daB das Auffinden von archàologischen Spuren der Charles-Kultur (ca. 6500-3200 Jahre v.u.Z.) eine Suche in tieferen Wassern erfordert

1,25-Dihydroxyvitamin D3 Enhances Bovine Mammary Epithelial Innate Immune Responses

Bovine mammary epithelial cells that were treated with bacterial lipopolysaccharide and 1,25-dihydroxyvitamin D3 showed an increase in the expression of the genes for inducible nitric oxide synthase (iNOS) and S100 calcium binding protein A12 (S100 A12). iNOS and S100 A12 are part of the innate immune response and expressed in the mammary gland during mastitis. Production of 1,25- dihydroxyvitamin D3 in the mammary gland during mastitis, then, may be an important component of the innate immune response

Regulation of Immune Responses to Mycobacteria bovis by a Paracrine Mechanism of Vitamin D Signaling in Cattle

We provide evidence that T-cell responses to Mycobacteria bovis are suppressed by the production of 1,25-dihydroxyvitamin D3 in monocytes and B-cells from cattle. Current vitamin D requirements for cattle are solely based on the classical endocrine mechanism of vitamin D signaling that regulates calcium homeostasis and should be re-evaluated to account for vitamin D signaling mechanisms in the immune system

Envelope protein complexes of Mycobacterium avium subsp. paratuberculosis and their antigenicity

Mycobacterium avium subsp. paratuberculosis (MAP) is the causative agent of Johne’s disease, a chronic enteric disease of ruminant animals. In the present study, blue native PAGE electrophoresis and 2D SDS-PAGE were used to separate MAP envelope protein complexes, followed by mass spectrometry (MS) to identify individual proteins within the complexes. Identity of individual proteins within complexes was further confirmed by MS upon excision of spots from 2D SDS-PAGE gels. Among the seven putative membrane complexes observed, major membrane protein (MAP2121c), a key MAP antigen involved in invasion of epithelial cells, was found to form a complex with cysteine desulfurase (MAP2120c). Other complexes found included those involved in energy metabolism (succinate dehydrogenase complex) as well as a complex formed by Cfp29, a characterized T cell antigen of Mycobacterium tuberculosis. To determine antigenicity of proteins, Western blot was performed on replicate 2D SDS-PAGE gels with sera from noninfected control cows (n = 9) and naturally infected cows in the subclinical (n = 10) and clinical (n = 13) stages of infection. Clinical animals recognized MAP2121c in greater proportion than subclinical and control cows, whereas cysteine desulfurase recognition was not differentiated by infection status. To further characterize antigenicity, recombinant proteins were expressed for 10 of the proteins identified and evaluated in an interferon-gamma (IFN-g) release assay as well as immunoblots. This study reveals the presence of protein complexes in the cell envelope of MAP, suggesting protein interactions in the envelope of this pathogen. Furthermore the identification of antigenic proteins with potential as diagnostic targets was characterized

MicroRNA regulation of bovine monocyte inflammatory and metabolic networks in an in vivo infection model.

peer-reviewedBovine mastitis is an inflammation-driven disease of the bovine mammary gland that costs the global dairy industry several billion dollars per annum. Because disease susceptibility is a multi-factorial complex phenotype, an integrative biology approach is required to dissect the molecular networks involved. Here, we report such an approach, using next generation sequencing combined with advanced network and pathway biology methods to simultaneously profile mRNA and miRNA expression at multiple time-points (0, 12, 24, 36 and 48h) in both milk and blood FACS-isolated CD14+ monocytes from animals infected in vivo with Streptococcus uberis. More than 3,700 differentially expressed (DE) genes were identified in milk-isolated monocytes (MIMs), a key immune cell recruited to the site of infection during mastitis. Up-regulated genes were significantly enriched for inflammatory pathways, while down-regulated genes were enriched for non-glycolytic metabolic pathways. Monocyte transcriptional changes in the blood, however, were more subtle but highlighted the impact of this infection systemically. Genes up-regulated in blood-isolated-monocytes (BIMs) showed a significant association with interferon and chemokine signalling. Furthermore, twenty-six miRNAs were differentially expressed in MIMs and three in BIMs. Pathway analysis revealed that predicted targets of down-regulated miRNAs were highly enriched for roles in innate immunity (FDR < 3.4E-8) in particular TLR signalling, while up-regulated miRNAs preferentially targeted genes involved in metabolism. We conclude that during S. uberis infection miRNAs are key amplifiers of monocyte inflammatory response networks and repressors of several metabolic pathways.This study was funded in part by Teagasc RMIS 6018 and United States Department of Agriculture ARS funding 3625-32000-102-00. NL is supported by a Teagasc Walsh Fellowship

Avian Intestinal Mucus Modulates Campylobacter jejuni Gene Expression in a Host-Specific Manner

Campylobacter jejuni is a leading cause of bacterial foodborne illness in humans worldwide. However, C. jejuni naturally colonizes poultry without causing pathology where it resides deep within mucus of the cecal crypts. Mucus may modulate the pathogenicity of C. jejuni in a species-specific manner, where it is pathogenic in humans and asymptomatic in poultry. Little is known about how intestinal mucus from different host species affects C. jejuni gene expression. In this study we characterized the growth and transcriptome of C. jejuni NCTC11168 cultured in defined media supplemented with or without mucus isolated from avian (chicken or turkey) or mammalian (cow, pig, or sheep) sources. C. jejuni showed substantially improved growth over defined media, with mucus from all species, showing that intestinal mucus was an energy source for C. jejuni. Seventy-three genes were differentially expressed when C. jejuni was cultured in avian vs. mammalian mucus. Genes associated with iron acquisition and resistance to oxidative stress were significantly increased in avian mucus. Many of the differentially expressed genes were flanked by differentially expressed antisense RNA asRNA, suggesting a role in gene regulation. This study highlights the interactions between C. jejuni and host mucus and the impact on gene expression, growth and invasion of host cells, suggesting important responses to environmental cues that facilitate intestinal colonization.IMPORTANCE Campylobacter jejuni infection of humans is an important health problem world-wide and is the leading bacterial cause of foodborne illnesses in U.S. The main route for exposure for humans is consumption of poultry meat contaminated during processing. C. jejuni is frequently found in poultry, residing within the mucus of the intestinal tract without causing disease. It is not clear why C. jejuni causes disease in some animals and humans, while leaving birds without symptoms. To understand its activity in birds, we characterized C. jejuni responses to poultry mucus to identify genes turned on in the intestinal tract of birds. We identified genes important for colonization and persistence within the poultry gut, turned on when C. jejuni was exposed to poultry mucus. Our findings are an important step in understanding how C. jejuni responds and interacts in the poultry gut, and may identify ways to reduce C. jejuni in birds

Treatment of an Intramammary Bacterial Infection with 25-Hydroxyvitamin D3

Deficiency of serum levels of 25-hydroxyvitamin D3 has been correlated with increased risk of infectious diseases such as tuberculosis and influenza. A plausible reason for this association is that expression of genes encoding important antimicrobial proteins depends on concentrations of 1,25-dihydroxyvitamin D3 produced by activated immune cells at sites of infection, and that synthesis of 1,25-dihydroxyvitamin D3 is dependent on the availability of 25-hydroxyvitamin D3. Thus, increasing the availability of 25(OH)D3 for immune cell synthesis of 1,25-dihydroxyvitamin D3 at sites of infection has been hypothesized to aid in clearance of the infection. This report details the treatment of an acute intramammary infection with infusion of 25-hydroxyvitamin D3 to the site of infection. Ten lactating cows were infected with in one quarter of their mammary glands. Half of the animals were treated intramammary with 25-hydroxyvitamin D3. The 25-hydroxyvitamin D3 treated animal showed significantly lower bacterial counts in milk and showed reduced symptomatic affects of the mastitis. It is significant that treatment with 25-hydroxyvitamin D3 reduced the severity of an acute bacterial infection. This finding suggested a significant non-antibiotic complimentary role for 25-hydroxyvitamin D3 in the treatment of infections in compartments naturally low in 25-hydroxyvitamin D3 such as the mammary gland and by extension, possibly upper respiratory tract infections

In Vivo Activation of the Intracrine Vitamin D Pathway in Innate Immune Cells and Mammary Tissue during a Bacterial Infection

Numerous in vitro studies have shown that toll-like receptor signaling induces 25-hydroxyvitamin D3 1α-hydroxylase (1α-OHase; CYP27B1) expression in macrophages from various species. 1α-OHase is the primary enzyme that converts 25-hydroxyvitamin D3 to 1,25-dihydroxyvitamin D3 (1,25(OH)2D3). Subsequently, synthesis of 1,25(OH)2D3 by 1α-OHase in macrophages has been shown to modulate innate immune responses of macrophages. Despite the numerous in vitro studies that have shown 1α-OHase expression is induced in macrophages, however, evidence that 1α-OHase expression is induced by pathogens in vivo is limited. The objective of this study was to evaluate 1α-OHase gene expression in macrophages and mammary tissue during an in vivo bacterial infection with Streptococcus uberis. In tissue and secreted cells from the infected mammary glands, 1α-OHase gene expression was significantly increased compared to expression in tissue and cells from the healthy mammary tissue. Separation of the cells by FACS9 revealed that 1α-OHase was predominantly expressed in the CD14+ cells isolated from the infected mammary tissue. The 24-hydroxylase gene, a gene that is highly upregulated by 1,25(OH)2D3, was significantly more expressed in tissue and cells from the infected mammary tissue than from the healthy uninfected mammary tissue thus indicating significant local 1,25(OH)2D3 production at the infection site. In conclusion, this study provides the first in vivo evidence that 1α-OHase expression is upregulated in macrophages in response to bacterial infection and that 1α-OHase at the site of infection provides 1,25(OH)2D3 for local regulation of vitamin D responsive genes

Rapid Reactivation of Extralymphoid CD4 T Cells during Secondary Infection

After infection, extralymphoid tissues are enriched with effector and memory T cells of a highly activated phenotype. The capacity for rapid effector cytokine response from extralymphoid tissue-memory T cells suggests these cells may perform a ‘sentinel’ function in the tissue. While it has been demonstrated that extralymphoid CD4+ T cells can directly respond to secondary infection, little is known about how rapidly this response is initiated, and how early activation of T cells in the tissue may affect the innate response to infection. Here we use a mouse model of secondary heterosubtypic influenza infection to show that CD4+ T cells in the lung airways are reactivated within 24 hours of secondary challenge. Airway CD4+ T cells initiate an inflammatory cytokine and chemokine program that both alters the composition of the early innate response and contributes to the reduction of viral titers in the lung. These results show that, unlike a primary infection, extralymphoid tissue-memory CD4+ T cells respond alongside the innate response during secondary infection, thereby shaping the overall immune profile in the airways. These data provide new insights into the role of extralymphoid CD4+ T cells during secondary immune responses

Vitamin D Signaling in the Bovine Immune System: A Model for Understanding Human Vitamin D Requirements

The endocrine physiology of vitamin D in cattle has been rigorously investigated and has yielded information on vitamin D requirements, endocrine function in health and disease, general metabolism, and maintenance of calcium homeostasis in cattle. These results are relevant to human vitamin D endocrinology. The current debate regarding vitamin D requirements is centered on the requirements for proper intracrine and paracrine vitamin D signaling. Studies in adult and young cattle can provide valuable insight for understanding vitamin D requirements as they relate to innate and adaptive immune responses during infectious disease. In cattle, toll-like receptor recognition activates intracrine and paracrine vitamin D signaling mechanism in the immune system that regulates innate and adaptive immune responses in the presence of adequate 25-hydroxyvitamin D. Furthermore, experiments with mastitis in dairy cattle have provided in vivo evidence for the intracrine vitamin D signaling mechanism in macrophages as well as vitamin D mediated suppression of infection. Epidemiological evidence indicates that circulating concentrations above 32 ng/mL of 25-hydroxyvitamin D are necessary for optimal vitamin D signaling in the immune system, but experimental evidence is lacking for that value. Experiments in cattle can provide that evidence as circulating 25-hydroxyvitamin D concentrations can be experimentally manipulated within ranges that are normal for humans and cattle. Additionally, young and adult cattle can be experimentally infected with bacteria and viruses associated with significant diseases in both cattle and humans. Utilizing the bovine model to further delineate the immunomodulatory role of vitamin D will provide potentially valuable insights into the vitamin D requirements of both humans and cattle, especially as they relate to immune response capacity and infectious disease resistance