Rough vaccines in animal brucellosis: structural and genetic basis and present status

Brucellosis control and eradication requires serological tests and vaccines. Effective classical vaccines (S19 in cattle and Rev 1 in small ruminants), however, induce antibodies to the O-polysaccharide of the lipopolysaccharide which may be difficult to distinguish from those resulting from infection and may thus complicate diagnosis. Rough attenuated mutants lack the O-polysaccharide and would solve this problem if eliciting protective immunity; the empirically obtained rough mutants 45/20 and RB51 have been used as vaccines. Strain 45/20 is reportedly unstable and it is not presently used. RB51 is increasingly used instead of S19 in some countries but it is rifampicin resistant and its effectiveness is controversial. Some controlled experiments have found good or absolute protection in adult cattle vaccinated orally (full dose) or subcutaneously (reduced dose) and in one field experiment, RB51 was reported to afford absolute protection to calves and to perform better than S19. Controlled experiments in calves, however, have shown reduced doses of RB51 to be ineffective, full doses only partially effective, and RB51 less effective than S19 against severe challenges. Moreover, other observations suggest that RB51 is ineffective when prevalence is high. RB51 is not useful in sheep and evidence in goats is preliminary and contradictory. Rough mutants obtained by molecular biology methods on the knowledge of the genetics and structure of Brucella lipopolysaccharide may offer alternatives. The B. abortus manBcore (rfbK) mutant seems promising in cattle, and analyses in mice suggest that mutations affecting only the O-polysaccharide result in better vaccines than those affecting both core and O-polysaccharide. Possible uses of rough vaccines also include boosting immunity by revaccination but solid evidence on its effectiveness, safety and practicality is not available.     

Haptoglobin and pig-major acute protein are increased in pigs with postweaning multisystemic wasting syndrome (PMWS)

The objective of this study was to determine the serum concentration levels of selected acute phase proteins (APP), haptoglobin (HPT) and pig-major acute phase protein (pig-MAP), in postweaning multisystemic wasting syndrome (PMWS) affected pigs and PCV2-subclinically infected pigs. In a first study, a group of 15 eight-week-old conventional pigs from a PMWS affected farm were bled and a complete necropsy, histopathology and in situ hybridisation to detect PCV2 were performed. Based on the results, pigs were classified as suffering from PMWS (n = 10) or healthy animals (n = 5). In a second study, a group of 45 pigs from another PMWS affected farm were selected and bled at 3, 7, 12 and 28 weeks of age. The assessment of PCV2 infection status in these pigs was retrospectively done by PCV2 PCR in serum samples. Selected APP were measured in the serum of all studied pigs by means of radial immunodiffusion. Mean HPT and pig-MAP levels were significantly increased (p = 0.004 and p = 0.0006 respectively) in PMWS-affected pigs when compared to levels found in healthy pigs (2.52 +/- 0.88 mg/mL vs. 1.06 +/- 0.73 mg/mL for HPT and 3.81 +/- 1.53 mg/mL vs. 0.76 +/- 0.34 mg/mL for pig-MAP). In the second study, no significant difference in mean HPT and pig-MAP values were observed between PCV2 PCR positive and negative pigs of any age. However, both APP increased significantly with age in PCV2 PCR negative pigs. Altogether, the present results suggest that APP levels are significantly increased in pigs that develop PMWS, but not in animals with a PCV2 subclinical infection.     

Use of volumetric capnography to identify pulmonary dysfunction in horses with and without clinically apparent recurrent airway obstruction

OBJECTIVE: To investigate whether volumetric capnography indices could be used to differentiate between horses without recurrent airway obstruction (RAO) and horses with RAO that were in clinical remission or that had clinically apparent RAO. ANIMALS: 70 adult Swiss Warmblood horses (20 used for pleasure riding and 50 used for dressage or show jumping). PROCEDURE: Horses were allocated to 4 groups on the basis of history, clinical signs, results of endoscopy, and cytologic findings (group 1, 21 healthy horses; group 2, 22 horses with RAO that were in remission; group 3, 16 horses with mild RAO; group 4, 11 horses with exacerbated RAO). Expiratory volume and CO2 curves were recorded by use of a computerized ultrasonic spirometer. Volumetric capnograms were plotted, and derived indices were calculated. RESULTS: Dead-space volume (VD) was calculated by use of the Bohr equation (VD(Bohr)) and for physiologic VD (VD(phys)). Ratios for VD(Bohr) to expiratory tidal volume (VT) and VD(phys) to V(T) as well as an index of effective CO2 elimination were significantly different among groups of horses. Age and use of the horses also significantly affected volumetric capnography indices. CONCLUSIONS AND CLINICAL RELEVANCE: Ratios of VD(Bohr) to VT and VD(phys) to VT as well as an index of effective CO2 elimination were sufficiently sensitive measures to distinguish between healthy horses and horses with RAO in remission. To optimize the ability of volumetric capnography indices to differentiate among horses in heterogeneous populations, it is important to account for effects of age and specific use of the horses.     

Crude protein fractions in common vetch (Vicia sativa L.) fresh forage during pod filling

Crude protein (CP) of forages can be separated into fractions of differentiated abilities to provide available amino acids in the lower gut of ruminants. This knowledge is critical to develop feeding systems and to predict animal responses. We have measured during two growing seasons (1996 to 1997 and 1997 to 1998) the CP fractions of common vetch fresh forage with the objective being to assess the influence of maturity on concentration of CP fractions (as a percentage of total CP) and fraction yields. Fraction B2, which represents true protein of intermediate ruminal degradation rate, was the largest single fraction in common vetch forage (about 40% of CP across seasons and maturity stages). Soluble fractions (A plus B1) were less than 50% of total CP while the unavailable fraction C ranged from 4 to 8% of total CP. As a result, the remaining fraction B3 (true protein of very low degradation rate) only represented 2 to 9% of total CP. Concentration and yield of fraction B3 increased (P < 0.05) from flowering to pod-filling. Results showed that undegraded dietary protein represented a small proportion of total CP in common vetch forage. Moving the harvesting stage from flowering to the pod filling phase allowed for greater yield of undegraded dietary protein.     

Muscle reorganisation through local injection of stem cells in the diaphragm of mdx mice

Background

The diaphragm is the major respiratory muscle affected by Duchenne muscular dystrophy (DMD) and is responsible for causing 80% of deaths. The use of mechanical forces that act on the body or intermittent pressure on the airways improves the quality of life of patients but does not prevent the progression of respiratory failure. Thus, diseases that require tissue repair, such as DMD, represent a group of pathologies that have great potential for cell therapy. The application of stem cells directly into the diaphragm instead of systemic application can reduce cell migration to other affected areas and increase the chances of muscle reorganisation. The mdx mouse is a suitable animal model for this research because its diaphragmatic phenotype is similar to human DMD. Therefore, the aim of this study was to assess the potential cell implantation in the diaphragm muscle after the xenotransplantation of stem cells.

Methods

A total of 9 mice, including 3 control BALB/Cmice, 3 5-month-old mdx mice without stem cell injections and 3 mdx mice injected with stem cells, were used. The animals injected with stem cells underwent laparoscopy so that stem cells from GFP-labelled rabbit olfactory epithelium could be locally injected into the diaphragm muscle. After 8 days, all animals were euthanised, and the diaphragm muscle was dissected and subjected to histological and immunohistochemical analyses.

Results

Both the fresh diaphragm tissue and immunohistochemical analyses showed immunopositive GFP labelling of some of the cells and immunonegativity of myoblast bundles. In the histological analysis, we observed a reduction in the inflammatory infiltrate as well as the presence of a few peripheral nuclei and myoblast bundles.

Conclusion

We were able to implant stem cells into the diaphragm via local injection, which promoted moderate muscle reorganisation. The presence of myoblast bundles cannot be attributed to stem cell incorporation because there was no immunopositive labelling in this structure. It is believed that the formation of the bundles may have been stimulated by cellular signalling mechanisms that have not yet been elucidated.     

Efficacy of alpha-cyclodextrin against experimental cryptosporidiosis in neonatal goats

The efficacy of orally administered tablets containing alpha-cyclodextrin, an excipient used in the pharmaceutical industry with demonstrated anticryptosporidial activity in vitro and in neonatal mice, was evaluated in neonatal goat kids. The formulation was evaluated for hardness and was subjected to in vitro drug release studies. Twenty goat kids were orally inoculated with 10(6) oocysts of C. parvum within the first 6 days of age. Half of the animals were treated by oral administration of four tablets of alpha-cyclodextrin/day (500 mg/kg of body weight) for six consecutive days, the treatment beginning on the day of inoculation. Infection was monitored by daily examination of faecal samples from the first day to 25 days post-inoculation. The criteria studied in evaluating efficacy were: oocyst shedding, presence of diarrhoea and weight gain at 15 and 25 days post-inoculation. alpha-cyclodextrin was effective when given at the beginning of infection: there was a longer pre-patent period, a reduction in the patent period and a decrease in the intensity of infection, these differences being statistically significant (P < 0.05) compared with untreated neonatal kids. Moreover, except in one animal, the diarrhoea was prevented in infected neonatal kids. Animals from both groups increased the body weight and no significant differences were seen between the two groups.