Lack of a type-2 glycosyltransferase in the fish pathogen Flavobacterium psychrophilum determines pleiotropic changes and loss of virulence

Flavobacterium psychrophilum is an important fish pathogen, responsible for Cold Water Disease, with a significant economic impact on salmonid farms worldwide. In spite of this, little is known about the bacterial physiology and pathogenesis mechanisms, maybe because it is difficult to manipulate, being considered a fastidious microorganism. Mutants obtained using a Tn4351 transposon were screened in order to identify those with alteration in colony morphology, colony spreading and extracellular proteolytic activity, amongst other phenotypes. A F. psychrophilum mutant lacking gliding motility showed interruption of the FP1638 locus that encodes a putative type-2 glycosyltransferase (from here on referred to as fpgA gene, Flavobacterium psychrophilum glycosyltransferase). Additionally, the mutant also showed a decrease in the extracellular proteolytic activity as a consequence of down regulation in the fpgA mutant background of the fpp2-fpp1 operon promoter, responsible for the major extracellular proteolytic activity of the bacterium. The protein glycosylation profile of the parental strain showed the presence of a 22 kDa glycosylated protein which is lost in the mutant. Complementation with the fpgA gene led to the recovery of the wild-type phenotype. LD₅₀ experiments in the rainbow trout infection model show that the mutant was highly attenuated. The pleiotropic phenotype of the mutant demonstrated the importance of this glycosyltranferase in the physiology and virulence of the bacterium. Moreover, the fpgA mutant strain could be considered a good candidate for the design of an attenuated vaccine.     

Acute sterile hemorrhagic cystitis after a single intravenous administration of cyclophosphamide in three dogs.

Three dogs receiving cyclophosphamide IV as part of a combination chemotherapeutic regimen developed macrohematuria, stranguria, and pollakiuria within 24 hours of administration of the first dose of this drug. An 11-year-old spayed mixed-breed dog with an oral squamous cell carcinoma was administered 250 mg of cyclophosphamide/m2 of body surface, whereas a 4-year-old castrated male Gordon Setter was treated with 100 mg of cyclophosphamide/m2 and a 6-year-old male German Shepherd Dog with a cutaneous hemangiosarcoma was administered 140 mg of cyclophosphamide/m2. Aerobic bacterial culture, antimicrobial susceptibility testing, and urinalysis were performed on urine obtained by cystocentesis from all 3 dogs after hematuria was observed. Sterile hemorrhagic cystitis was diagnosed on the basis of large numbers of RBC in the urine, lack of pathogens on bacterial culturing of urine, and clinical signs. Although cyclophosphamide-induced cystitis in dogs has been reported in the literature numerous times, acute episodes developing within 24 hours of administration of the first dose have not been reported in this species with the use of therapeutic doses. Therefore, appropriate precautionary steps should be taken, even when the drug is being administered intermittently.     

Pharmacokinetics of enrofloxacin in fingerling rainbow trout (Oncorhynchus mykiss)

The pharmacokinetics of intravenously and orally administered enrofloxacin was determined in fingerling rainbow trout (Oncorhynchus mykiss). Doses of 5 or 10 mg enrofloxacin/kg body weight were administered intravenously to 26 fish for each dose and blood was sampled over a 60-h period at 15 degrees C. Two groups of fish were treated orally with 5, 10, or 50 mg/kg (80 fish/dose at each temperature) and held at 15 degrees C or 10 degrees C during the 60-h sampling period. Following intravenous administration, the serum concentration-time data of enrofloxacin in rainbow trout were best described by a two-compartment open model for both doses of 5 and 10 mg enrofloxacin/kg. The hybrid rate constants alpha and beta did not differ between doses. The distributional phase was rapid with a half-life of 6-7 min for both doses. Overall half-lives of elimination were 24.4 h (95% CI = 20.2-30.8) and 30.4 h (24.2-41.0), respectively, for the 5- and 10-mg/kg doses. A large Vd(area) was observed following dosing of either 5 or 10 mg enrofloxacin/kg,: 3.22 and 2.56 l/kg, respectively. Whole body clearance for 5 mg/kg was 92 ml/h.kg and 58 ml/h.kg at the 10-mg/kg dose. Following oral administration, the serum concentration-time data for enrofloxacin were best described as a one-compartment open model with first-order absorption and elimination. Apparent Ka over all doses at 10 degrees C averaged 62% less than apparent Ka at 15 degrees C. Estimates of the apparent t(1/2)e over both temperatures ranged from 29.5 h (18.4-73.4) to 56.3 h (38.3-106.6). Bioavailability averaged 42% over all doses at 15 degrees C and was decreased to an average of 25% at 10 degrees C. Peak serum concentrations appeared between 6 and 8 h following dosing. A dose of 5 mg/kg/day was estimated to provide average steady-state serum concentrations at 10 degrees C that are approximately 4.5 times the highest reported MIC values for Streptococcus spp., the fish pathogen least sensitive to enrofloxacin. Owing to the long apparent half-life of elimination of enrofloxacin in fingerling trout, it would take approximately 5 to 9 days to achieve these predicted steady-state serum concentrations; this estimate is important when considering the duration of therapy in clinical trials.     

Site and extent of neutral detergent fiber digestion, efficiency of ruminal digesta flux and fecal output as related to variations in voluntary intake and chewing behavior in heifers

Six heifers (1/4 Brahman x 1/4 Jersey x 1/2 Angus, 290 kg avg wt) with ruminal, duodenal and ileal cannulas were given ad libitum access to a corn silage-based diet with or without 100 mg monensin.hd-1.d-1 in a two-period crossover design. There was no significant effect of monensin on voluntary intake (VI), flow of NDF to the abomasum, fecal output or the efficiency of transfer of NDF to the duodenum (unitary fluxes of NDF through the reticulo-omasal orifice, number.g NDF.(kg BW.75)-1). Monensin increased extent of NDF digestion (NDFD) in the dorsal (P = .07) and ventral (P = .09) ruminal sac. As a fraction of the total tract NDFD, NDFD increased progressively in dorsal rumen (.214), ventral rumen (.548) and duodenal (.818) sites. The VI among individual heifers was negatively related to unitary fluxes of NDF through the reticulo-omasal orifice (P = .001) and to the extent of digestion of potentially digestible NDF (PDNDFD) in the ventral rumen (P = .006) and duodenum (P = .032), but not in the dorsal rumen (P = .720) or in the total tract (P = .785). Fecal output of NDF was negatively related to unitary times of eating (P = .027), ruminating (P = .001), masticating (P = .001) and unitary number of rumination boli (P = .001). These relationships all were influenced (P = .001) by individual heifer.(ABSTRACT TRUNCATED AT 250 WORDS)     

Natural infections of pigs with bovine viral diarrhoea virus associated with signs resembling swine fever

Between 1976 and 1985 while using immunofluorescence in the laboratory diagnosis of swine fever (SF), 13 incidents of bovine viral diarrhoea virus (BVDV) infection were detected in the Netherlands. Ultimate differentiation between swine fever virus (SFV) and BVDV was based on herd evidence supported by comparative antibody studies on sera from pigs inoculated with the isolate and from contact pigs in the herd of origin, using reference strains of SFV and BVDV. Recently differentiation of SFV and BVDV has been facilitated by typing the isolates with monoclonal antibodies. Signs suspicious of SF were observed in pigs two to 16 weeks old and were always confined to animals of one litter. In most cases the affected litter died out gradually although once an animal recovered. Stillbirth and neonatal death as well as the late onset of disease and its limitation to a single litter in a herd suggested a congenital route of infection. Although transplacental infection of BVDV in pigs has been reported before, these cases are believed to be the first in which natural BVDV infections could be associated with clinical signs and pathological lesions indistinguishable from those observed in chronic SF.     

Congenital inguinal hernias associated with a rent in the common vaginal tunic in five foals

Five foals were admitted because of colic that developed within 24 hours after birth. Physical examination revealed swelling from the inguinal region to the cranial aspect of the prepuce. Through the skin overlying the swelling, distended loops of bowel could be palpated. The hernias could be reduced with the foals restrained in dorsal recumbency. Surgical exploration showed the swelling to be an inguinal hernia with small bowel herniation through a rent in the common vaginal tunic. Concurrent rupture of the vaginal tunic should be suspected when a congenital inguinal hernia is associated with colic, swelling that extends to the prepuce, or intestine that is easily palpated within the hernial swelling. Repeated manual reduction in these cases would be of little benefit and surgical intervention would be recommended.     

Anthelmintic efficacies of fenbendazole, ivermectin, and levamisole against Nematodirus battus infections in lambs

Anthelmintic efficacies of fenbendazole, ivermectin, and levamisole were evaluated against naturally acquired infections of Nematodirus battus in lambs. Four groups of 10 lambs each were used. Oral administration of 8 mg of levamisole/kg of body weight or 5 mg of fenbendazole/kg significantly (P less than 0.01) reduced the degree of infection by N battus (adults) by greater than 99%. An oral formulation of 200 micrograms of ivermectin/kg was 98% effective (P less than 0.01). Numbers of other Nematodirus spp (including N filicollis and N spathiger) were significantly reduced.     

A comparison of three methods of end-to-end anastomosis in the equine small colon

Six animals were used to compare simple interrupted, simple continuous, and stapled techniques for end-to-end rotated small colon anastomoses. Three ponies were evaluated three days after surgery for adhesion formation and luminal diameter at each anastomosis site. Two ponies and one horse were evaluated 14 days after surgery for adhesion formation. All anastomosis sites were examined histologically for alignment of tissue planes, and evidence of inflammation. No single technique was superior in all areas examined. While only three of the nine anastomoses had mild fibrinous adhesions at three days after surgery, all anastomoses had varying degrees of well-organized adhesions by 14 days after surgery. Three days after surgery luminal diameter was most comprised by the simple continuous technique. Although the simple interrupted technique was most consistent in its alignment of tissue planes, it was only slightly better than the simple continuous pattern. Stapled anastomoses were inverted or everted at all sites examined. Adhesion formation was less extensive, and histologic evidence of inflammation was less severe, in stapled anastomoses. The average times required to complete the simple interrupted, simple continuous, and stapled anastomoses were 22, 14, and 8 minutes, respectively.     

Pre-weaning supplementary feed and porcine post-weaning diarrhoea

Attempts were made to induce an intestinal hypersensitivity response to weaner diet by feeding pigs with small quantities of this material before weaning. In two trials using different weaner diets piglets subjected to this regimen showed no significant differences in small intestinal structure, in ability to absorb xylose, in bodyweight gain, in incidence of diarrhoea or excretion of enteropathogens after weaning compared with pigs not given any of the diet before weaning, or fed with a different diet before weaning. When post-weaning diarrhoea occurred it was associated with an earlier, more prolonged and greater proliferation of enterotoxigenic Escherichia coli in the small intestines than occurred in healthy pigs after weaning. The greater proliferation in pigs which developed diarrhoea could not be attributed either to an excessive dietary intake after weaning, or to a specific proliferation of rotaviruses.     

Some pharmacokinetic data of aditoprim and trimethoprim in healthy and tick-borne fever infected dwarf goats

Aditoprim (AP) is a new dihydrofolate reductase inhibitor, which is structurally related to trimethoprim (TMP). The pharmacokinetics of AP (10 mg/kg) and TMP (20 mg/kg) were assessed in healthy dwarf goats. Therapeutic efficacy against rickettsial infections was tested in tick-borne fever (TBF) infected goats. The animals were given TMP (n = 5) or AP (n = 5) by i.v. injection, and subsequently the drugs were administered orally (same groups, similar doses). Finally, both groups were infected with TBF and the i.v. experiment was repeated. Plasma concentration-time curves for both drugs followed first-order two-compartment decay. For TMP, mean t1/2 beta +/- SEM (h) was 0.84 +/- 0.06 (i.v. control) and 0.90 +/- 0.06 (i.v. infected), respectively, whereas for AP values of 8.00 +/- 0.31 (i.v. control) and 10.28 +/- 0.67 (i.v. infected) were obtained (P less than 0.05). Mean Vd beta +/- SEM values (l/kg) were 3.84 +/- 0.27 (i.v. control) and 4.07 +/- 0.85 (i.v. infected) for TMP (NS) and 7.02 +/- 0.63 vs 9.29 +/- 0.21 (P less than 0.05) for AP. After i.v. injection, rumen fluid concentrations of AP were significantly (P less than 0.05) higher and more persistent than those of TMP. For AP, the plasma and rumen fluid concentrations at 3 h were 1.20 +/- 0.06 micrograms/ml and 0.85 +/- 0.17 microgram/ml, respectively. After oral administration of TMP, Cmax in plasma was 0.12 +/- 0.01 microgram/ml and the maximum was reached after 1.2 +/- 0.16 h; systemic bioavailability (F) was 10.3% (relative to AUC i.v.). Oral treatment with AP resulted in a Cmax value of 0.21 +/- 0.02 microgram/ml with Tmax of 22.5 +/- 1.65 h and a F value of 71%. Based on WBC, serum ALP and rectal temperature responses, it was concluded that both TMP and AP were inactive against Ehrlichia phagocytophila.