The transition between infection of the mammalian host and colonization of an arthropod vector is required for the ongoing transmission of a broad array of pathogens, from viruses to protozoa. Understanding how this transition is mediated provides opportunities to disrupt transmission through either chemotherapy or immunization. We used an unbiased proteomic screen to identify Anaplasma marginale proteins specifically upregulated in the tick compared to the mammalian host. Comparative mass spectrometric analysis of proteins separated by two-dimensional gel electrophoresis of uninfected and infected ISE6 cells and infected mammalian cells identified 15 proteins exclusively expressed or upregulated in tick cells. All 15 had originally been annotated as hypothetical proteins. We confirmed quantitative upregulation and expression in situ within the midgut epithelial and salivary gland acinar cells of vector ticks during successful transmission. The results support the hypothesis that A. marginale gene expression is regulated by the specific host environment and, in a broader context, that the core genome evolved in the arthropod vector with differential regulation, allowing adaptation to mammalian hosts. Furthermore, the confirmation of the in situ expression of candidates identified in ISE6 cell lines indicates that this approach may be widely applicable to bacteria in the genera Anaplasma and Ehrlichia, removing a major technical impediment to the identification of new targets for vaccine and chemotherapeutic blocking of transmission.The transition between infection of the mammalian host and colonization of an arthropod vector is required for ongoing transmission of a broad array of pathogens, from viruses to protozoa. Understanding how this transition is mediated provides opportunities to disrupt transmission through either chemotherapy or immunization. Bacteria in the genera Anaplasma and Ehrlichia are obligate intracellular pathogens and effectively invade, survive, and replicate in markedly different cell types in the mammalian host and ixodid ticks, the arthropod vector (4). Impressively, this transition is effected by using a small genome of Ͻ1.5 Mb (2,3,8,9,15). We and others have hypothesized that the bacterial proteome would be specifically molded for each environment, with a core set of proteins expressed universally and subsets specifically up-or downregulated depending on the host/vector environment (6,12,19,26,27). However, there has been only minimal proteomic evidence that supports accepting this hypothesis. The best evidence comes from recent analysis of E. chaffeensis that detected proteins present in either in vitro-infected tick cells or canine macrophages (26); however, unique or upregulated expression of these candidate proteins in the tick cells has not been confirmed. There has been no identification of bacterial proteins specifically upregulated or exclusively expressed during actual colonization in the tick.We addressed this knowledge gap by an unbiased proteomic approach using the St. Mar...

Aims: The aim of the study was to determine Campylobacter jejuni contamination and prevalence on fomites moving between broiler farms and the processing plant in the period after cleaning and before departure to harvest chickens. In addition, changes in the proportion of contaminated fomites in the course of a day were assessed. Methods and Results: Pooled swab samples were obtained from pallets, crates, wheels of trucks, tractors and forklifts, truck beds, and from drivers' and catchers' boots. After enrichment in Bolton's broth Campylobacter were recovered on modified blood-free Campylobacter selective agar (mCCDA). Isolates were identified using tests for phenotypic and biochemical characteristics. Of the 209 samples collected, 53% were positive for C. jejuni, with all fomites positive except tractor wheels. Pallets had the highest contamination rate at 75%. More than 50% of catchers' boots, drivers' boots, crates and truck wheels were positive. Forty-seven per cent and 31% of truck beds and forklift wheels, respectively, were contaminated. The proportion of contaminated fomites did not change significantly during the day. Conclusions: This study has identified trucks, forklifts, pallets, crates, drivers' and catchers' boots as potential sources of C. jejuni for broilers. Significance and Impact of the Study: Campylobacter jejuni contamination of broiler processing plant fomites was found to be extensive ranging from 31% for truck beds to 75% for pallets. The proportion of contaminated fomites was observed to be similar throughout the day. The impact of contaminated fomites as sources of colonization of broilers with C. jejuni is discussed.

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Transmission of tick-borne pathogens requires transition between distinct host environments with infection and replication in host-specific cell types. Anaplasma marginale illustrates this transition: in the mammalian host, the bacterium infects and replicates in mature (nonnucleated) erythrocytes, while in the tick vector, replication occurs in nucleated epithelial cells. We hypothesized that proteins containing ankyrin motifs would be expressed by A. marginale only in tick cells and would traffic to the infected host cell nucleus. A. marginale encodes three proteins containing ankyrin motifs, an AnkA orthologue (the AM705 protein), AnkB (the AM926 protein), and AnkC (the AM638 protein). All three A. marginale Anks were confirmed to be expressed during intracellular infection: AnkA is expressed at significantly higher levels in erythrocytes, AnkB is expressed equally by both infected erythrocytes and tick cells, and AnkC is expressed exclusively in tick cells. There was no evidence of any of the Ank proteins trafficking to the nucleus. Thus, the hypothesis that ankyrin-containing motifs were predictive of cell type expression and nuclear localization was rejected. In contrast, AnkA orthologues in the closely related A. phagocytophilum and Ehrlichia chaffeensis have been shown to localize to the host cell nucleus. This difference, together with the lack of a nuclear localization signal in any of the AnkA orthologues, suggests that trafficking may be mediated by a separate transporter rather than by endogenous signals. Selection for divergence in Ank function among Anaplasma and Ehrlichia spp. is supported by both locus and allelic analyses of genes encoding orthologous proteins and their ankyrin motif compositions.