Subdivision of Salmonella enterica serovar enteritidis phage types PT14b and PT21 by plasmid profiling

We have shown that plasmid profiling is a sensitive method for further identification of strains of Salmonella enterica serovar enteritidis (S. enteritidis) phage type PT21 and to a lesser extent the strains of phage type PT14b. Five and three plasmid types were identified within 33 strains of phage type PT21 and 19 strains of phage type PT14b, respectively. Plasmid types in strains of phage type PT21 showed significant correlation with geographical origin of the strain. In strains of phage type PT14b a single isolate predominated suggesting that the plasmid designated as 'C' can be directly linked with S. enteritidis PT14b strains. Application of IS200 fingerprinting did not reveal any other differences and showed just one copy of IS200 in all the 52 analysed strains. All the strains were tested for antibiotic resistance and only four strains were resistant to ampicillin, cefotaxime, cefuroxime and cotrimoxazole. This indicates that low molecular weight plasmids in Salmonella enteritidis are not responsible for the spread of antibiotic resistance.     

Influences of diet and vaccination on colonisation of pigs by the intestinal spirochaete Brachyspira (Serpulina) pilosicoli

The purpose of this study was to determine whether methods used to control swine dysentery (SD), caused by the intestinal spirochaete Brachyspira (Serpulina) hyodysenteriae, would also be effective in controlling porcine intestinal spirochaetosis (PIS) caused by the related spirochaete Brachyspira (Serpulina) pilosicoli. Weaner pigs in Groups I (n=8) and II (n=6) received a standard weaner pig diet based on wheat and lupins, whilst Group III (n=6) received an experimental diet based on cooked white rice and animal protein. Pigs in Group II were vaccinated intramuscularly twice at a 3-week-interval with a formalinised bacterin made from B. pilosicoli porcine strain 95/1000 resuspended in Freund's incomplete adjuvant. Eleven days later pigs in all groups were infected orally with 10(10) cells of strain 95/1000 on three successive days. One control pig in Group I developed acute diarrhoea, and at post-mortem had a severe erosive colitis with end-on attachment of spirochaetes to the colonic epithelium. All other pigs developed transient mild diarrhoea and had moderate patchy colitis at post-mortem 3 weeks later. B. pilosicoli was isolated from the faeces of all pigs, except for one fed rice, and was isolated from the mesenteric nodes of three pigs from Group I and from one vaccinated pig in Group II. Consumption of the rice-based diet, but not vaccination, delayed and significantly (p<0.001) reduced the onset of faecal excretion of B. pilosicoli after experimental challenge. Vaccination induced a primary and secondary serological response to B. pilosicoli, as measured using sonicated whole cells of strain 95/1000 as an ELISA plate coating antigen. Antibody titres in the vaccinated pigs then declined, despite intestinal colonisation by B. pilosicoli. Both groups of unvaccinated animals also failed to develop a post-infection increase in circulating antibody titres.     

Involvement of CD28/CTLA4-B7 costimulatory pathway in the development of lymphadenopathy and splenomegaly in MRL/lpr mice

MRL/lpr mouse is an established animal model which develops autoimmune diseases including glomerulonephritis, sialoadenitis, hepatitis and inflammatory lung disease. Additionally, it has been reported that lpr strains uniquely accumulate CD3+ CD4- CD8- B220+ (double negative, DN) T cells in lymphoid organs leading to lymphadenopathy and splenomegaly. To investigate the role of CD28/CTLA4-B7 pathway in the development of lymphadenopathy and splenomegaly, MRL/lpr mice were treated with soluble form of CTLA4 molecules, CTLA4IgG, which efficiently blocks this pathway. It was demonstrated that (i) the development of DN T cells was independent of the CD28/CTLA4-B7 pathway, (ii) the CD28/CTLA4-B7 pathway was required for the development of lymphadenopathy and splenomegaly, (iii) the CD28/CTLA4-B7 pathway was important for the accumulation of various cell populations in the lymph node and spleen, (iv) composition of the accumulating cell populations was not altered by CTLA4IgG treatment, and (v) activation of conventional T cells and IL-4 production from conventional T cells were the CD28/CTLA4-B7 pathway dependent. Thus, we concluded that the CD28/CTLA4-B7 pathway was required for the development of full-blown lymphadenopathy and splenomegaly in MRL/lpr mice.     

Congenital thymic aplasia in a Holstein-Israeli female calf

An unusual case of congenital thymic aplasia in a Holstein-Israeli female calf is described. The most prominent clinical findings were diarrhoea and weakness. At necropsy, the only significant pathological finding was the marked decrease in thymus size. Histologically, this organ was composed of loose connective tissue in which a few lymphocytes were scattered randomly.     

Disseminated mycoses in a dog by Paecilomyces sp

We describe a case of canine mycoses initially diagnosed by clinical signs and enzyme-linked immunosorbent assay anti-fungal test, and later confirmed by the isolation of Paecilomyces sp. during the post-mortem examination. The fungus was isolated from lesions in the kidneys, mitral valve, abdominal aorta and vertebral discs. In this kind of process, it is important to identify the responsible agent early in order to make a study of anti-fungal susceptibility and establish effective treatment.     

The distribution of vitamin A and retinol-binding protein in the blood plasma, urine, liver and kidneys of carnivores

The contents of retinol and retinyl esters as well as retinol-binding protein (RBP) in the plasma, urine, liver and kidneys of dogs, raccoon dogs and silver foxes were investigated. In the plasma and urine of all three species, vitamin A was present as retinol and retinyl esters. Vitamin A levels (1376+/-669 microg x g(-1)) were significantly higher in the livers of dogs than in the kidneys (200+/-217 microg x g(-1), P < 0.001 ). However, vitamin A levels in the kidneys of raccoon dogs (291+/-146 microg x g(-1)) and silver foxes (474+/-200 microg x g(-1)) were significantly higher than in the liver (67+/-58 microg x g(-1) and 4.3+/-2.4 microg x g(-1), respectively, both P < 0.001). RBP was immunologically detected in the blood plasma of all species, but never in the urine. In the liver, immunoreactive RBP was found in hepatocytes. In the kidneys of all species, RBP was observed in the cells of the proximal convoluted tubules. The levels of vitamin A in the livers of raccoon dogs and silver foxes were extremely low, which would be interpreted as a sign of great deficiency in humans. This observation might indicate that the liver status cannot be used as an indicator of vitamin A deficiency in canines. The high levels of vitamin A in the kidneys in all three species may indicate a specific function of the kidney in the vitamin A metabolism of canines.     

Morphology and amplitude values of the electrocardiogram of Spanish-bred horses of different ages in the Dubois leads system

The aim of this work was to record the modifications of the form and amplitude of the electrocardiographic tracings during growth using the Dubois system and to study its sensibility to these physiological changes. This work was carried out on 179 healthy, young and adult Spanish-bred horses (98 females and 81 males). One electrocardiogram (ECG) was obtained from each horse using the Dubois leads system. The bifid shape deflection of the P wave was generally more frequent than the simple one in animals at 4 months of age or older. Amplitudes of the P2 component and of the simple positive wave presented significant differences (p < 0.01) between the age groups studied: animals up to 3 months in age and older than 2 years of age (highest amplitudes), and the rest of the animals (lowest amplitudes). Significant variations (p < 0.05) were found for the QS amplitude between animals of 1 month of age, and all the other groups. The multiple range analysis did not define a clear distribution of age groups for the amplitude of the T wave. It may therefore be concluded, that in the Spanish-bred horse up to 1 month of age, the form and amplitude of the QRS complex in the Dubois leads system differ from those of older animals. Thus, this study shows the sensibility of the Dubois leads system for detecting the electrocardiographic changes related to the growing process in the Spanish-bred horse. In addition, it provides standard values of electrocardiographic parameters in the Spanish-bred horse at different stages of growth.     

In vitro isolation and characterization of bovine Neospora caninum in Korea

The brains of nine aborted bovine fetuses and two newborn calves born from dams suspected to be infected with Neospora caninum were homogenized and inoculated into Vero cells. All fetuses and calves were from cows determined as seropositive to N. caninum by an IFA test. Sera and thoracic fluids of all fetuses and calves also revealed high antibody titer to N. caninum by IFAT ranging from 1:800 to 1:3200. N. caninum was isolated from the brains of one aborted fetus and one newborn calf when the brain homogenates were grown continuously in Vero cell culture. N. caninum tachyzoites, giemsa-positive, were first observed on Days 45 and 56 postinoculation in the newborn calf and the aborted fetus, respectively. The isolates (KBA-1 and KBA-2) were morphologically and ultrastructurally similar to previously published Neospora isolates. The isolated parasites were confirmed as N. caninum by means of the antigenic reactivities, immunostaining, PCR and southern blotting, and electron microscopy.     

Can an increased intake of metabolizable protein affect the periparturient relaxation in immunity against teladorsagia circumcincta in sheep?

Within the overall hypothesis that the periparturient relaxation of immunity (PPRI) to gastrointestinal parasites has a nutritional basis, we studied whether an increased intake of metabolizable protein (MP) could affect the expression of immunity to Teladorsagia (Ostertagia) circumcincta of twin-bearing and -rearing Greyface ewes. Two sets of iso-energetic diets were formulated to provide either 85% (L-) or 130% (H-) of the MP requirements during the last 6 weeks of pregnancy (-P) and the first 6 weeks of lactation (-L). Thirty ewes were allocated to LP or HP. Six ewes of each group were slaughtered after lambing (S(0)); the remaining 24 ewes of each group were allocated to either LL or HL, thus creating four treatment combinations during lactation (LP/LL, LP/HL, HP/LL and HP/HL; n=12). Six ewes of each treatment combination were slaughtered at Week 3 (S(3)) or at Week 6 (S(6)) in lactation.The HP ewes were heavier at lambing, produced heavier lambs, had lower faecal egg counts (FECs), a lower concentration of plasma pepsinogen, a higher concentration of plasma albumin during late pregnancy and more abomasal globule leukocytes (GLs) but similar worm burdens at S(0) compared to the LP ewes. The LP/HL and HP/HL ewes had a higher calculated milk production than the LP/LL and HP/LL ewes. FECs increased for each treatment combination during the first 3 weeks of lactation. This increase concurred with fewer abomasal GLs at S(3) compared to S(0). The LP/HL and HP/HL ewes had a higher proportion of immature worms and more abomasal GLs than the LP/LL and HP/LL ewes at S(6). The FECs continued to increase during the second 3 weeks of lactation for the LP/LL ewes only, which concurred with an elevated plasma pepsinogen concentration and an absence of abomasal GLs at S(6). The plasma pepsinogen concentration decreased rapidly for the LP/HL ewes during lactation. We conclude that an increased intake of MP could enhance the expression of immunity to T. circumcincta of twin-bearing and -rearing Greyface ewes.     

Safety of selamectin in dogs

Selamectin is a broad-spectrum avermectin endectocide for treatment and control of canine parasites. The objective of these studies was to evaluate the clinical safety of selamectin for topical use in dogs 6 weeks of age and older, including breeding animals, avermectin-sensitive Collies, and heartworm-positive animals. The margin of safety was evaluated in Beagles, which were 6 weeks old at study initiation. Reproductive, heartworm-positive, and oral safety studies were conducted in mature Beagles. Safety in Collies was evaluated in avermectin-sensitive, adult rough-coated Collies. Studies were designed to measure the safety of selamectin at the recommended dosage range of 6-12mgkg(-1) of body weight. Endpoints included clinical examinations, clinical pathology, gross and microscopic pathology, and reproductive indices. Selected variables in the margin of safety and reproductive safety studies were subjected to statistical analyses. Pups received large doses of selamectin at the beginning of the margin of safety study when they were 6 weeks of age and at their lowest body weight, yet displayed no clinical or pathologic evidence of toxicosis. Similarly, selamectin had no adverse effects on reproduction in adult male and female dogs. There were no adverse effects in avermectin-sensitive Collies or in heartworm-positive dogs. Oral administration of the topical formulation caused no adverse effects. Selamectin is safe for topical use on dogs at the recommended minimum dosage of 6mgkg(-1) (6-12mgkg(-1)) monthly starting at 6 weeks of age, and including dogs of reproducing age, avermectin-sensitive Collies, and heartworm-positive dogs.