According to Bergey's Manual of Systematic Bacteriology, the Rickettsiales are '...bacteria with typical Gram-negative cell walls and no flagella'. The recently sequenced genome of 'Candidatus Midichloria mitochondrii', a divergent lineage within the order Rickettsiales capable of invading mitochondria in ixodid ticks, revealed the presence of 26 putative flagellar genes. Open questions in relation to this observation are whether these genes are expressed and whether they possess the domains expected for the flagellar function. Here we show that: (a) the putative flagellar proteins of 'Ca. M. mitochondrii' actually possess the conserved domains and structural features required for their function in a model bacterium; (b) the seven flagellar genes of 'Ca. M. mitochondrii' that have been tested are expressed at the RNA level; and (c) the putative flagellar cap gene of this bacterium (FliD) is expressed at the protein level, and can be stained within the bacterium and at its surface. Beside the specific questions that we have addressed that relate to the first evidence, to our knowledge, for a flagellar apparatus in a member of the order Rickettsiales, we present here novel tools (recombinant protein and antibodies) that will facilitate the study of 'Ca. M. mitochondrii'.
In Chile, the horn fly, Hematobia irritans (L., 1758), is a major pest of grazing cattle and affects livestock production during the summer. Previous studies in Europe and the United States have shown that cattle flies, including H. irritans, are differentially attracted to individual cattle within herds and that volatile semiochemicals are responsible for this phenomenon. This study provides evidence that similar differential attractiveness occurs for the interaction between Chilean Holstein-Friesian cattle herds and local H. irritans populations. Thus, Holstein-Friesian dairy cattle, Bos taurus, which were of similar age and physiological condition, were shown to possess an uneven distribution of H. irritans. Heifers h6904 and h8104 were defined as low-carrier heifers and h5804, h2304 and h1404 as high-carrier heifers. Gas chromatography (GC) and coupled GC-mass spectrometric (GC-MS) analysis of samples collected from heifers revealed the presence of compounds previously reported as semiochemicals for cattle flies, including meta- and para-cresol, methylketones (C8-C11), and 6-methyl-5-hepten-2-one. Other compounds identified included carboxylic acids (butanoic, 3-methylbutanoic, pentanoic, and hexanoic acids), 1-hexanol, and 3-octanone. In Y-tube olfactometer studies, both m- and p-cresol attracted H. irritans at the highest doses tested (10(-6) g), as did the positive control 1-octen-3-ol. Of the other compounds tested, only 2-decanone and 2-undecanone produced a behavioral response, with significantly more flies being recorded in the control arm when the former compound was tested (at 10(-6) and 10(-8) g), and more flies being recorded in the treated arm for the latter compound (at 10(-7) g). This demonstration of behavioral activity with the identified compounds represents a first step for research into the application of semiochemicals in monitoring and control of cattle flies in Chile.
Thirty-one faecal samples were collected from red deer in the northern area of Varese, in the Italian region of Lombardy, between August and October 2008. The animals had either been hunted or accidently killed. Examination for internal parasites showed a prevalence of 45.2% for Elaphostrongylus cervi larvae and species identification was confirmed by polymerase chain reaction (PCR). Ninety-seven faecal samples were also collected from two goat flocks grazing in the same area between December 2007 and May 2008. These showed a prevalence of 74.7% for lungworms. Furthermore, the central nervous systems from five goats and one sheep from this area with a history of neurologically related lameness were examined. Histopathology confirmed E. cervi cerebro-spinal nematodiasis in five cases out of six. This study demonstrates E. cervi transmission from wild to domestic ruminants when the animals graze in the same area, and the possible occurrence of clinical disease in infected goats and sheep associated with high prevalence in deer.
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