Consensus statement on ehrlichial disease of small animals from the infectious disease study group of the ACVIM. American College of Veterinary Internal Medicine

Within the past several decades, the number of Ehrlichia spp. recognized to infect cats, dogs, and human beings has expanded substantially. The recent application of advanced techniques in molecular biology has changed how ehrlichiosis is diagnosed and has provided new tools for the assessment of treatment. As these techniques are applied, the numerous questions that relate to the management of dogs and cats with ehrlichiosis ultimately will be answered. We hope this consensus statement will assist veterinarians in the management of their patients.     

Characterization of,and immune responses of mice to,the purified OmpA-equivalent outer membrane protein of Pasteurella multocida serotype A:3 (Omp28)

Pasteurella multocida A:3 is a major cause of bovine pneumonia. A major antigenic heat-modifiable 28kDa outer membrane protein (Omp28) was previously identified. The purpose of this study was to purify and characterize Omp28 immunologically and structurally. Omp28 was extracted from N-lauroylsarcosine-insoluble protein preparations by a combination of detergent fractionation with Zwittergent 3-14 and chromatography. Partial N-terminal amino acid sequence confirmed Omp28 as a member of the OmpA-porin family. However, porin activity could not be demonstrated in a lipid-bilayer assay. Heat modifiability of purified Omp28 was demonstrated, and Omp28 was found in outer membrane fraction of P. multocida. Surface exposure of Omp28 was demonstrated by partial protease digestion of intact bacteria, by binding of anti-Omp28 polyclonal ascites fluid to the bacterial surface, and by partial inhibition of anti-outer membrane antiserum binding by previous incubation of the bacteria with anti-Omp28 serum. CD-1 mice vaccinated with purified Omp28 developed a significant antibody titer (P<0.05) compared to the control treatment group but were not protected from a homologous intraperitoneal bacterial challenge. By contrast, treatment groups vaccinated with P. multocida outer membrane, formalin-killed P. multocida or a commercial vaccine were significantly protected from challenge. In vitro complement-mediated killing of P. multocida was observed in post-vaccination sera of outer membrane, formalin-killed P. multocida, and commercial vaccine-treatment groups, but not with sera from the Omp28-treatment group. In conclusion, although Omp28 is surface exposed and antigenic, it may not be a desirable immunogen for stimulating immunity to P. multocida.     

Effect of different fat sources on in vitro degradation of nutrients and certain blood parameters in sheep

This study was designed to determine the effects of calcium salt of palm oil fatty acids (CS), hydroxyethylsoyamide (HESA), butylsoyamide (BSA) and soybean oil (SO) on degradation of crude protein and fibre in vitro, and on the blood plasma lipid parameters in vivo. Five mature wethers (body weight 75 kg) were fed five diets in a 5 x 5 Latin square experiment. The control diet consisted of 50% meadow hay and 50% concentrate with no added fat. The control diet was supplemented with CS, HESA, BSA, or SO. Fat was added at 3.5% of dietary dry matter (DM). The final ether extract content of the ration was near 6%. Each period lasted 20 days. Fat supplements, except HESA, consistently decreased the in vitro DM disappearance of soybean meal as compared to control. In contrast to the effect of other treatments, crude protein degradation was greatest in the test tubes with inocula obtained from sheep fed diet with HESA. Fat supplements equally inhibited the DM and fibre breakdown of alfalfa pellet. CS and HESA seemed to be less detrimental to in vitro fermentation of neutral detergent fibre (NDF) than BSA and SO. All fat supplements increased blood plasma triglyceride, cholesterol and total lipid content. Plasma concentration of cholesterol and total lipid was highest with SO. The inclusion of CS in the diet increased 16:0, while all fat supplements increased plasma 18:0 and decreased 16:1 and 18:1 fatty acid content. Plasma 18:2n-6 was not changed by feeding CS and SO. However, compared to the control diet, 18:2n-6 increased with 12 and 41% in plasma fatty acids when sheep were fed HESA and BSA, respectively. The results showed that plasma concentration of linoleic acid was enhanced more when the amide was synthesised from butylamine than when from ethanolamine.     

Mice immune responses to Brucella abortus heat shock proteins. Use of baculovirus recombinant-expressing whole insect cells,purified Brucella abortus recombinant proteins,and a vaccinia virus recombinant as immunogens

Brucella abortus resists the microbicidal mechanisms of macrophages, and the expression of its heat shock proteins (HSPs) such as GroEL, GroES and HtrA may play a role in this resistance. Bacterial HSPs can be very immunogenic, inducing protective immunity in various types of bacterial infections. However, the significance of immune responses directed against B. abortus HSPs in the protection against brucellosis is currently unresolved. To elucidate the role of these proteins in protection against Brucella challenge, individual, divalent or trivalent baculovirus (BV) recombinants of B. abortus GroEL, GroES and/or HtrA were injected into BALB/c mice either as protein-expressing whole cells or as purified proteins. The preparations were given to mice in combination with Freund's or Ribi adjuvant, respectively. In addition, some mice were primed with a vaccinia virus-GroEL recombinant, followed by inoculation with purified GroEL-Ribi adjuvant combination. Antibodies were observed against B. abortus GroEL and HtrA, but not against GroES. Cellular immune response was demonstrated by observing significant IFN-gamma release by lymphocytes of mice immunized with the purified HtrA-Ribi adjuvant combination. However, none of the mice inoculated with individual, divalent or trivalent HSP-expressing cells combined with complete Freund's adjuvant or inoculated with purified B. abortus HSPs combined with Ribi adjuvant, were protected against challenge with B. abortus virulent strain 2308. Priming with vaccinia virus-GroEL recombinant and boosting with GroEL-Ribi combination did not induce protective immunity. Based on the results obtained, we suggest that although humoral and cell-mediated immune responses are induced, but protective immune response is not induced by B. abortus HSPs.     

Pituitary effects of steroid hormones on secretion of follicle-stimulating hormone and luteinizing hormone

Steroid hormones have a profound influence on the secretion of the gonadotropins, follicle-stimulating hormone (FSH) and luteinizing hormone (LH). These effects can occur as a result of steroid hormones modifying the secretion of gonadotropin-releasing hormone (GnRH) from the hypothalamus, or a direct effect of steroid hormones on gonadotropin secreting cells in the anterior pituitary gland. With respect to the latter, we have shown that estradiol increases pituitary sensitivity to GnRH by stimulating an increase in expression of the gene encoding the GnRH receptor. Since an estrogen response element (ERE) has not been identified in the GnRH receptor gene, this effect appears to be mediated by estradiol stimulating production of a yet to be identified factor that in turn enhances expression of the GnRH receptor gene. However, the importance of estradiol for enhancing pituitary sensitivity to GnRH during the periovulatory period is questioned because an increase in mRNA for the GnRH receptor precedes the pre-ovulatory rise in circulating concentrations of estradiol. In fact, it appears that the enhanced pituitary sensitivity during the periovulatory period may occur as a result of a decrease in concentrations of progesterone rather than due to an increase in concentrations of estradiol. Estradiol also is capable of altering secretion of FSH and LH in the absence of GnRH. In a recent study utilizing cultured pituitary cells from anestrous ewes, we demonstrated that estradiol induced a dose-dependent increase in secretion of LH, but resulted in a dose-dependent decrease in the secretion of FSH. We hypothesized that the discordant effects on secretion of LH and FSH might arise from estradiol altering the production of some of the intrapituitary factors involved in synthesis and secretion of FSH. To examine this hypothesis, we measured amounts of mRNA for activin B (a factor known to stimulate synthesis of FSH) and follistatin (an activin-binding protein). We found no change in the mRNA for follistatin after treatment of pituitary cells with estradiol, but noted a decrease in the amount of mRNA for activin B. Thus, the inhibitory effect of estradiol on secretion of FSH appears to be mediated by its ability to suppress the expression of the gene encoding activin.     

Clinical Leptospira interrogans serogroup Australis serovar lora infection in a stud farm in the Netherlands

Summary

A Leptospira interrogans serogroup australis serovar lora infection in a stud farm is reported. During three successive years (1984–1986) clinical leptospirosis with a severe often rapid, fatal course was seen in 12 foals.

Clinical examination revealed severe respiratory distress, depression and pyrexia. Other symptoms were diarrhea (2), jaundice (1), and an unsteady gait (1). Morphological characteristics of the disease were massive pulmonary haemorrhage and haemorrhagicthrombotic or extracapillary glomerulonephritis with tubulonephrosis and interstitial oedema. In most foals high or increasing MAT titres to serovar bratislava were found; from one foal Leptospira interrogans serovar lora was isolated.

Serological examination of all 56 mares at the farm (August 1986) revealed antibodies to serovar bratislava in 64 per cent of the animals. These findings support the idea that Leptospira interrogans serovar bratislava and closely related strains (in this study serovar lora) may be adapted to and maintained by the horse population.