The laboratory evaluation of abdominal, thoracic, and pericardial effusions is a useful diagnostic tool for the assessment of disease states that result in fluid accumulation. Although the numeric ...values pertaining to cell count and protein content are important, the microscopic evaluation is a critical aspect of the diagnostic procedure; not only does it allow complete classification of the fluid but it allows identification of specific cell types or microorganisms that might be responsible for the fluid accumulation. These findings should always be interpreted in conjunction with the history, signalment, physical findings, and other diagnostic aids in making a definitive diagnosis.
Organisms in the family
Anaplasmataceae are important tick-borne pathogens of livestock worldwide and cause recently emergent infections in humans. Despite their medical importance, very little is ...known about how these organisms regulate gene expression in the mammalian host, the tick vector, or during transition between the host and vector. However, it is clear that gene regulation, in addition to recombinatorial mechanisms, is essential for these small genome pathogens to adapt to distinctly different environments. In this study, we identify and establish the function of three promoter elements in the locus encoding major outer membrane protein expression sites in both
Anaplasma marginale and
Anaplasma phagocytophilum. Gene expression from this locus involves both classical and atypical polycistronic transcripts. The identified promoter elements have a structure similar to that defined in
Escherichia coli and are functional in driving protein expression in a prokaryotic cell-free transcription and translation system and in recombinant
E. coli. The two strongest promoters identified in vitro and with recombinant
E. coli were also shown to be functional in
A. marginale infected cells, as determined by quantification of downstream transcripts. The promoters in both
A. marginale and
A. phagocytophilum have similar structure and activity, supporting the conclusion that the two loci are syntenic with conservation of function. In addition, they share structural elements within the promoters that appear to be likely sites for regulation. These data enhance our understanding of how expression of these variable outer membrane proteins may be controlled in the key stages of tick-borne transmission and infection.
Anaplasma marginale has recently been shown to infect endothelial cells in vitro, but it remains unknown as to whether endothelial infection also occurs in vivo. In this report, we demonstrate ...through dual fluorescence microscopy that A marginale, detected by the monoclonal antibody ANAF16C1, co-localizes with the endothelial cell marker, von Willebrand factor, in tissue sections from an experimentally inoculated calf. The results indicate that A marginale infection includes endothelial cells and has implications for both pathogenesis and immune mechanisms.
Dogs acquire infections with the Anaplasmataceae family pathogens, E. canis, E. chaffeensis, E. ewingii, A. platys and A. phagocytophilum mostly during summer months when ticks are actively feeding ...on animals. These pathogens are also identified as causing diseases in people. Despite the long history of tick-borne diseases in dogs, much remains to be defined pertaining to the clinical and pathological outcomes of infections with these pathogens. In the current study, we performed experimental infections in dogs with E. canis, E. chaffeensis, A. platys and A. phagocytophilum. Animals were monitored for 42 days to evaluate infection-specific clinical, hematological and pathological differences. All four pathogens caused systemic persistent infections detectible throughout the 6 weeks of infection assessment. Fever was frequently detected in animals infected with E. canis, E. chaffeensis, and A. platys, but not in dogs infected with A. phagocytophilum. Hematological differences were evident in all four infected groups, although significant overlap existed between the groups. A marked reduction in packed cell volume that correlated with reduced erythrocytes and hemoglobin was observed only in E. canis infected animals. A decline in platelet numbers was common with E. canis, A. platys and A. phagocytophilum infections. Histopathological lesions in lung, liver and spleen were observed in all four groups of infected dogs; infection with E. canis had the highest pathological scores, followed by E. chaffeensis, then A. platys and A. phagocytophilum. All four pathogens induced IgG responses starting on day 7 post infection, which was predominantly comprised of IgG2 subclass antibodies. This is the first detailed investigation comparing the infection progression and host responses in dogs after inoculation with four pathogens belonging to the Anaplasmataceae family. The study revealed a significant overlap in clinical, hematological and pathological changes resulting from the infections.
To evaluate light microscopic, cytochemical, and ultrastructural characteristics of blood cells from eastern diamondback rattlesnakes.
10 healthy snakes.
Various stains, including Wright-Giemsa, ...benzidine peroxidase, Sudan black B, chloroacetate esterase, alpha-naphthyl butyrate esterase, acid phosphatase, leukocyte alkaline phosphatase, periodic acid-Schiff with diastase, and toluidine blue, were used to stain leukocytes differentially on multiple blood smears. Electron microscopy also was performed.
Lymphocytes were the most commonly observed leukocyte and could be distinguished from thrombocytes, using periodic acid-Schiff stain with diastase. Azurophils also were commonly observed; their granules stained with peroxidase. Eosinophils were not identified; however, 2 morphologic variations of heterophils were seen in the blood of all snakes and were considered the same cell type at different stages of cytoplasmic granule development. Heterophil granules were better preserved, using a one-step Wright-Giemsa method that did not require alcohol fixation prior to staining. Degranulated heterophils were observed in all preparations.
Most leukocytes of eastern diamondback rattlesnakes can be identified easily on Wright-Giemsa-stained preparations. However, hematologic stains that do not require alcohol fixing prior to staining may be preferred for leukocyte evaluation in certain reptiles. A limited degree of heterophil maturation may continue in the blood of healthy snakes. This, along with degranulation of heterophils, may result in a variable staining pattern in this cell type, regardless of the stain used.
Results provide baseline data for use in hematologic testing in diagnosis of disease and monitoring of treatment of sick or injured snakes.
To determine clinical signs, results of diagnostic testing, underlying cause, and outcome in cats with seizures.
Retrospective study.
17 cats with seizures.
Only those cats in which an underlying ...metabolic abnormality causing the seizures had been identified, diagnostic imaging of the brain and CSF analysis had been done, or a necropsy had been performed were included. Seizures were classified as being a result of metabolic disease, symptomatic epilepsy (ie, epilepsy resulting from a structural lesion of the brain), or probably symptomatic epilepsy (ie, epilepsy without any extracranial or identifiable intracranial disease that is not suspected to be genetic in origin).
3 cats had seizures associated with an underlying metabolic disease (hepatic encephalopathy), 7 had symptomatic epilepsy (3 with neoplasia and 4 with meningoencephalitis), and 7 had probably symptomatic epilepsy. Six of the 7 cats with symptomatic epilepsy died or were euthanatized within 3 months after the diagnosis was made, whereas 6 of the 7 cats with probably symptomatic epilepsy survived for at least 12 months after the diagnosis was made.
Results suggest that cats with probably symptomatic epilepsy may have a good long-term prognosis.
Antisera against different Ehrlichiae recognize an immunodominant, cross-reacting ∼28 kDa surface antigen defined as the MAP1 in
Cowdria ruminantium.These antigens are considered valuable in ...developing serodiagnostic tests and recombinant vaccines for Ehrlichiae infections. To evaluate the relationship in three closely related Ehrlichiae,
Ehrlichia chaffeensis, Ehrlichia canis,and
C. ruminantium,the structure of the 28 kDa antigen genes was analyzed. We describe the cloning and characterization of DNA encoding genes homologous to MAP1 from
E. chaffeensisand
E. canis.The cloned segment of
E. chaffeensiscontains one expressed and four transcriptionally silent tandemly arranged, nonidentical genes; the
E. canislocus consists of two nonidentical genes. Comparative analysis of these genes revealed the presence of four conserved regions separated by three highly variable regions. B-cell epitope analysis identified three major cross-reacting epitopes that map to the variable regions. Location of the epitopes at the variable regions and the presence of multigene family with only one expressed copy suggest a mechanism of immune evasion in these Ehrlichiae.
The ecological success of social insects is based on division of labour, not only between queens and workers, but also among workers. Whether a worker tends the brood or forages is influenced by age, ...fertility and nutritional status, with brood carers being younger, more fecund and more corpulent. Here, we experimentally disentangle behavioural specialization from age and fertility in Temnothorax longispinosus ant workers and analyse how these parameters are linked to whole‐body gene expression. A total of 3,644 genes were associated with behavioural specialization which is ten times more than associated with age and 50 times more than associated with fertility. Brood carers were characterized by an upregulation of three Vitellogenin (Vg) genes, one of which, Vg‐like A, was the most differentially expressed gene that was recently shown experimentally to control the switch from brood to worker care. The expression of Conventional Vg was unlinked to behavioural specialization, age or fertility, which contrasts to studies on bees and some ants. Diversity in Vg/Vg‐like copy number and expression bias across ants supports subfunctionalization of Vg genes and indicates that some regulatory mechanisms of division of labour diverged in different ant lineages. Simulations revealed that our experimental dissociation of co‐varying factors reduced transcriptomic noise, suggesting that confounding factors could potentially explain inconsistencies across transcriptomic studies of behavioural specialization in ants. Thus, our study reveals that worker gene expression is mainly linked to the worker's function for the colony and provides novel insights into the evolution of sociality in ants.
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