Arachidonic acid and prostaglandin E2 in Malpighian tubules of female yellow fever mosquitoes.

The fatty acid compositions of Malpighian tubules from adult females of the mosquito Aedes aegypti were determined for total lipids, phospholipids, triacylglycerols and three phospholipid fractions, namely phosphatidylcholine (PC), phosphatidylethanolamine (PE) and phosphatidylinositol/phosphatidylserine (PI/PS). The prostaglandin precursor arachidonic acid (20:4n-6) occurred in total lipids and phospholipids, but not triacylglycerols. Within phospholipids, nearly all of the 20:4n-6 was detected in PC, with only traces in PE, and none was detected in PI/PS. Isolated Malpighian tubules incorporated exogenous radioactive 20:4n-6 into tissue phospholipids and diacylglycerols, with most of the radioactivity recovered in diacylglycerol. These data indicate selective incorporation of 20:4n-6 into tissue lipids. PGE2 was detected in Malpighian tubule whole mounts by immunohistochemical staining. These findings support the idea that prostaglandins are physiologically active in mosquito Malpighian tubules.

Cell membrane permeability and mitochondrial dysfunction-inducing activities in cell-free supernatants from Serpulina hyodysenteriae serotypes 1 and 2.

Membrane permeability (MP) and mitochondrial dysfunction-inducing (MDI) activities were detected in cell-free supernatants (CFS) of Serpulina hyodysenteriae, using either hemoglobin release from porcine red blood cells (RBC) or cytoplasmic lactate dehydrogenase release from porcine peripheral blood lymphocytes (PBL), and reduction of the 3-(4,5-dimethylthiazoyl-2-y1)-2,5-diphenyltetrazolium bromide dye by porcine PBL. The MP and MDI activities of CFS correlated with each other for serotype 1 and 2 isolates taken at different population densities; however, the kinetics of toxin production varied between each serotype. The loss of enteropathogenicity of two field isolates with nonpathogenic phenotypes and pathogenic isolates passaged up to 45 times in vitro was not attributable to a loss of either membrane permeability or mitochondrial dysfunction-inducing activity of cell-free supernatants. Results from this study suggested the potential for two separate toxins being involved in the pathogenesis of swine dysentery, with the MDI activity correlating with age susceptibility to clinical disease.

Motility-regulated mucin association of Serpulina pilosicoli, the agent of colonic spirochetosis of humans and animals.

Colonic spirochetosis is a disease of humans and animals characterized by colonization of the colonic mucus gel and intimate attachment of Serpulina pilosicoli to the apical membrane of enterocytes. Motility-regulated mucin association plays a key role in colonic infection by the related spirochete Serpulina hyodysenteriae, the cause of swine dysentery. In this study the chemotaxis of Serpulina pilosicoli porcine isolate P43/6/78, human isolate SP16, and canine isolate 16242-94 was examined by anaerobic incubation of each spirochete in control medium or medium containing increasing concentrations of D-L serine or porcine gastric mucin (PGM). The porcine isolate had a chemotactic response towards 10 mM D-L serine, but not towards PGM. By contrast, the human and canine isolates were attracted towards 0.1% PGM, but not towards DL-serine. The composition of the growth medium appeared to modulate the chemotactic response of S. pilosicoli towards PGM; the loss of a chemotactic response of spirochetes grown in medium without pig fecal extract was restored by growing the spirochetes in medium containing 0.1% PGM. Serpulina pilosicoli displays a chemotactic response towards PGM which is modulated by the presence of certain substrate during the growth phase of the spirochete.

Recovery from colonic infection elicits serum IgG antibodies to specific Serpulina pilosicoli outer membrane antigens (SPOMA).

Colonic spirochetosis caused by S. pilosicoli is a disease of human and animals characterized by intimate attachment of the spirochete to colonic epithelial cells and colitis. To identify antigens that are potentially involved in recovery from the disease, whole-cell lysate (WC) and various detergent extracts including Sarkosyl-soluble (SS) and insoluble (SI), and Triton X-114 detergent phase (TXD) and aqueous phase (TXA) of the human isolate SP16 were examined by Western blotting with Serpulina spp. periplasmic flagellar protein FlaB-specific monoclonal antibody 7G2 as well as pooled pre-immune serum (PS), hyperimmune serum (HS), and convalescent serum (CS) from swine. The HS reacted with several antigens that were not identified by the CS, including the periplasmic flagellar proteins and some lower molecular weight bands. The CS identified three major immunoreactive double (D) or single (S) bands of approximately: (i) 64-kDa in the WC(S), SS(D), and TXD/A(S), (ii) 54-kDa in the WC(S), SS/I(S), and TXD(S), and (iii) 47-kDa in the SS(S) fraction. The data indicate recovery from colonic infection elicits serum IgG antibodies to specific S. pilosicoli outer membrane antigens (SPOMA).