Oral vaccination of brushtail possums with BCG: Investigation into factors that may influence vaccine efficacy and determination of duration of protection

AIMS: To determine factors that may influence the efficacy of an oral pelleted vaccine containing Mycobacterium bovis bacille Calmette-Guérin (BCG) to induce protection of brushtail possums against tuberculosis. To determine the duration of protective immunity following oral administration of BCG.

METHODS: In Study 1, a group of possums (n=7) was immunised by feeding 10 pellets containing dead Pasteur BCG, followed 15 weeks later with a single pellet of live Pasteur BCG. At that time, four other groups of possums (n=7 per group) were given a single pellet of live Pasteur BCG orally, a single pellet of live Danish BCG orally, 10 pellets of live Pasteur BCG orally, or a subcutaneous injection of live Pasteur BCG. For the oral pelleted vaccines, BCG was formulated into a lipid matrix, and each pellet contained approximately 10⁷ colony forming units (cfu) of BCG, while the vaccine injected subcutaneously contained 10⁶ cfu of BCG. A sixth, non-vaccinated, group (n=7) served as a control. All possums were challenged by the aerosol route with a low dose of virulent M. bovis 7 weeks after vaccination, and killed 7–8 weeks after challenge. Protection against challenge with M. bovis was assessed from pathological and bacteriological findings.

In Study 2, lipid-formulated live Danish BCG was administered orally to three groups of possums (10–11 per group), and these possums were challenged with virulent M. bovis 8, 29 or 54 weeks later. The possums were killed 7 weeks after challenge, to assess protection in comparison to a non-vaccinated group.

RESULTS: The results from Study 1 showed that vaccine efficacy was not adversely affected by feeding dead BCG prior to live BCG. Feeding 10 vaccine pellets induced a level of protection similar to feeding a single pellet. Protection was similar when feeding possums a single pellet containing the Pasteur or Danish strains of BCG. All vaccinated groups had significantly reduced pathological changes or bacterial counts when compared to the non-vaccinated group. In Study 2, oral administration of Danish BCG induced protection against challenge with M. bovis, which persisted for at least 54 weeks after vaccination. Some protection was observed in possums challenged 54 weeks after vaccination, but this protection was significantly less than that observed in groups vaccinated 29 or 8 weeks prior to challenge. There was a strong relationship between the proportion of animals producing positive lymphocyte proliferation responses to M. bovis antigens and protection against challenge with M. bovis.

CONCLUSIONS: Factors considered potentially capable of interfering with vaccination, including feeding dead BCG to possums prior to feeding live BCG, feeding multiple doses of BCG at one time, and changing strains of BCG, were shown not to interfere with the acquisition of protective immune responses in possums. Protection against tuberculosis was undiminished up to 29 weeks after vaccination with BCG administered orally. It is concluded that vaccination of possums by feeding pellets containing BCG is a robust and efficient approach to enhance the resistance of these animals to tuberculosis.     

Effectiveness of a non-surgical alternative to the Mules operation in sheep

Objective To measure changes to the perineal bare area, local tissue reaction and healing responses of young sheep, following intradermal administration of cetrimide and polyvinylpyrrolidone (PVP), with and without ethanol, to the breech and tail. Method A needle‐less injector was used to deposit formulations containing 40 g/L cetrimide and 30 g/L PVP (group 2) or 20 g/L cetrimide, 30 g/L PVP and 15 g/L ethanol (group 3), within the dermis of the tail and the region surrounding the perineal bare breech area of groups (N = 8) of Merino weaner sheep. The dimensions of the perineal bare area (length, width and diagonal distances left and right) and tail width were recorded before and at intervals after treatment for 60 days. Observations of swelling and bruising and scab formation at the treatment sites were recorded for up to 35 days after treatment. Rectal temperatures were monitored for up to 35 days after treatment and bodyweight for up to 60 days after treatment. An untreated control group (group 1) was included. Results Comparison of day ‐3 and day 35 measurement data showed that both treated groups had significantly (P < 0.05) wider breech bare areas compared to the untreated controls and that group 2 sheep had significantly (P < 0.05) longer breech bare areas compared to group 3 sheep or to the untreated controls, which were not significantly different. At this time scabs were still firmly in place on many treated sheep. At day 35 there was no increase in tail bare area caused by either treatment. By day 60 there was no significant difference between the treated and control groups in either the breech or tail regions indicating that the changes present at day 35, were not permanent. Mean weight gain in the groups throughout the 60‐day interval was unaffected by treatment. Intradermal treatment was associated with a significant elevation in body temperature. This effect lasted for 3 days and was associated with signs of discomfort and depressed appearance in at least some of the treated sheep. Bruising was mild to severe in all treated sheep within two days of treatment but was not evident in any sheep by day 21. Mild to moderate swelling was also associated with treatment but was not uniform across sheep in the groups. The tail of one sheep was severely swollen for several days. Swelling remained obvious in most treated sheep until day 14 but was not present at day 21. Conclusion Under the conditions of this study intradermal injection of cetrimide had no permanent effect on bare area measurements on the breech or the amount of wool‐bearing skin on the tail. It also caused signs of discomfort and pain that raise welfare concerns.     

Serological analysis of Chlamydophila abortus in Australian sheep and implications for the rejection of breeder sheep for export

Objective   To examine the incidence of positive results in a complement fixation test (CFT) and enzyme-linked immunosorbent assay (ELISA) for Chlamydophila abortus in Australian sheep and how this incidence differs with state of origin, age, sex, breed and property. To examine the consequences in relation to rejection of breeder sheep for export.
Design   Collection of blood samples from 891 sheep on 109 properties in southern Australia. All samples had a unique, coded property identification.
Procedure   The samples were tested using the Institut Pourquier Chlamydophila abortus antibody ELISA (rELISA) and a CFT. Residual maximum likelihood analyses of the sample to positive ratio of the corrected optical density for the rELISA and generalised linear mixed model analyses of the CFT outcomes were carried out.
Results   The sample to positive ratio of the corrected optical density values of the rELISA did not differ between sex, age, breed or state of origin, but differed greatly between properties. The CFT outcome did not differ between age, breed or state of origin, but differed greatly between properties and was more often positive with rams than with ewes.
Conclusion   Positive outcomes to C. abortus antibody tests are very common in Australia. Rams have a particularly high incidence of positive results with the CFT. Rejection of sheep and property consignments is likely to be very common with all tests and situations examined except for the CFT (at 1:32 dilution) in ewes.     

Spatial Variability of Turbulent Mixing in the Abyssal Ocean

Ocean microstructure data show that turbulent mixing in the deep Brazil Basin of the South Atlantic Ocean is weak at all depths above smooth abyssal plains and the South American Continental Rise. The diapycnal diffusivity there was estimated to be less than or approximately equal to 0.1 x 10(-4) meters squared per second. In contrast, mixing rates are large throughout the water column above the rough Mid-Atlantic Ridge, and the diffusivity deduced for the bottom-most 150 meters exceeds 5 x 10(-4) meters squared per second. Such patterns in vertical mixing imply that abyssal circulations have complex spatial structures that are linked to the underlying bathymetry.     

Use of enrofloxacin for treatment of large-form Haemobartonella felis in experimentally infected cats

OBJECTIVE: To compare treatment with enrofloxacin and doxycycline with no treatment in cats experimentally infected with Haemobartonella felis. DESIGN: Prospective case-control study. ANIMALS: 16 cats. PROCEDURE: Cats were inoculated with large-form H. felis from a chronically infected donor. Cats were assigned to 1 of 4 treatment groups: doxycycline (5 mg/kg [2.3 mg/lb], p.o., q 12 h), low-dose enrofloxacin (5 mg/kg, p.o., q 24 h), high-dose enrofloxacin (10 mg/kg [4.5 mg/lb], p.o., q 24 h), and an untreated control group. Clinical signs, Hct, blood smears, and a polymerase chain reaction (PCR) assay were used to monitor progression of the infection. RESULTS: All cats were confirmed to be infected with H. felis via blood smear evaluations and PCR assay results. Treatment had no effect on Hct during the intratreatment period, but Hct values were significantly greater in the low-dose enrofloxacin group, compared with the control group, during the posttreatment period. During the intratreatment period, H. felis organism counts per 1,000 RBC in the doxycycline treatment and the high-dose enrofloxacin treatment groups decreased at a significantly faster rate than those in the control group. In the posttreatment period, organism counts in the doxycycline treatment group and the low- and high-dose enrofloxacin groups decreased at significantly faster rates than counts in the control group. There was no significant effect of treatment on the number of positive PCR assay results. Two cats treated with enrofloxacin and 1 cat treated with doxycycline completely cleared the H. fe is organism despite presumed immunosuppression caused by glucocorticoids. CONCLUSIONS AND CLINICAL RELEVANCE: Results support the hypothesis that enrofloxacin has anti-H. felis effects.     

Malignant Sertoli Cell Tumour in a Young Simmenthal Bull – Clinical and Pathological Observations

Tumours are rare in the bovine testicle. A case of malignant Sertoli cell tumour in a 29‐month‐old Simmenthal bull that was hospitalized with a history of severe unilateral scrotal swelling is reported. On inspection and palpation, the scrotal sac was found enlarged with fluctuant content in the right side. The right testicle was enlarged, hard and indolent. Also the right plexus pampiniformis and funiculus spermaticus were enlarged. Sonograms revealed severe changes in the right testicle with a loss of homogeneity and multiple hyperechogenic areas. After slaughter, the scrotum with testicles were removed and evaluated pathologically. On section, the right testicle contained areas of necrosis, haemorrhage, and mineralization. Histology showed Sertoli cells in tubular structures surrounded by dense fibrous stroma replacing normal testicular tissue. Both lymphatic and blood vessels were infiltrated by neoplastic cells. Immunohistochemically, the neoplastic cells stained positive for vimentin and negative for cytokeratin and S‐100. Based on the pathological observations a diagnosis of right‐sided malignant Sertoli cell tumour with vascular invasion and hydrocele was established.