Luminol-dependent chemiluminescence analysis of the variables of the phagocytic response by canine granulocytes

Variables which influence the oxygen-dependent chemiluminescence (CL) response of canine polymorphonuclear leukocytes (PMN) to zymosan were examined in a luminol-dependent CL assay system. Maximal CL responses were obtained when 5 × 10⁶ canine PMN, isolated from heparinized blood, were assayed at 37° C in a Luminometer. The response was enhanced by the addition of 0.05 mM Luminol and inhibited by the addition of 0.06 mM sodium azide and 60 ug superoxide dismutase. Repeatability on a given day was very good; however, day to day variations in CL activity prevented direct comparison of phagocytic activity between days. Opsonization of zymosan in equine serum significantly reduced the CL response by canine PMN as compared to opsonization of zymosan in autologous or homologous canine serum and bovine serum. The present results show that luminol-dependent CL analysis can be used to measure phagocytosis by canine granulocytes in a luminometer and has potential use in clinical situations.     

Cell-mediated immune response after Brucella abortus S19 vaccination.

Cell-mediated immunity to Brucella abortus S19 vaccine was measured in young heifers by the microassay for stimulation of protein synthesis (SPS) with [3H]leucine and the skin test for delayed hypersensitivity. Brucella melitensis protein allergen and a crude B abortus S19-soluble antigen were compared in the SPS test. The SPS test was negative in 5 unvaccinated heifers and strongly positive in 3 twice-vaccinated steers. However, the SPS test was positive only in 13 of 30 S19-vaccinated heifers and the delayed hypersensitivity in 9 of 29 S19-vaccinated heifers. The 2 tests gave good agreement. Vaccination-induced residual antibody titers were partly correlated with the outcome of the tests used to measure cell-mediated immunity.     

Serum antibody response to soluble extract of the third-larval stage of Ostertagia ostertagi in cattle

Serum IgG, IgM, and IgA antibody responses against L₃ antigens of Ostertagia ostertagi were monitored by enzyme-linked immunosorbent assay (ELISA) after one, two or multiple sequential inoculations of this nematode in calves. Following the first infection, antibody levels did not change. After a second inoculation, IgG increased significantly (P < 0.05) after 2 months. IgG was not significantly increased 1 month after challenge inoculation. IgM and IgA antibody levels did not change following the first or second inoculations of L₃. IgG antibody levels rose only slightly following multiple sequential inoculations with infectious L₃.

Results indicate that calves with ostertagiasis have very weak serum antibody responses to L₃, and these appear to be of little value in detection of the infection in these animals.     

Flavobacterium columnare genomovar influences mortality in channel catfish (Ictalurus punctatus)

Flavobacterium columnare, causal agent of columnaris disease, is pathogenic to many species of freshwater fish throughout the world. The United States channel catfish (Ictalurus punctatus) aquaculture industry is severely impacted by columnaris disease. The majority of the F. columnare isolates recovered from diseased channel catfish belonged to either genomovars I or II. The objective of the present study was to determine if differences existed in the ability of these genomovars to induce mortality in channel catfish. Single strand conformation polymorphism analysis (SSCP) was used to ascribe the isolates used in this study to the appropriate genomovar. Immersion challenge experiments (15min immersion exposure to approximately 5x10(5) to 1x10(6) CFU/mL) were carried out to assess virulence of genomovar I and II isolates to channel catfish. The results demonstrated that genomovar II (n=4) isolates were significantly (P<0.05) more virulent to channel catfish fry (92-100% mortality) than genomovar I (n=3) isolates (0-46% mortality). In vivo adhesion of the genetically characterized F. columnare also correlated (r2=0.73) to increased mortality in the challenged fry. In fingerling channel catfish, significantly higher mortality (P<0.05) resulted with genomovar II isolates ALM-05-182 and ALG-00-530 as compared to all the genomovar I isolates (n=3). Mortality of genomovar II isolate BGFS-27 with similar to genomovar II isolate (ALG-00-530) and two genomovar I isolates (ALM-05-53 and 140). The results suggest that although both genomovars are present in the aquatic environment, genomovar II appears to be more pathogenic for channel catfish.     

Thiamin Requirement of Nile Tilapia,Oreochromis niloticus

Juvenile Nile tilapia, Oreochromis niloticus, were fed to apparent satiation twice daily with purified diets containing 0, 1.0, 2.0, 4.0, 8.0, and 16.0 mg/kg and 0, 2.0, 4.0, 8.0, 16.0, and 32.0 mg/kg of thiamin in separate 14‐ and 8‐wk trials (Experiments 1 and 2, respectively). Fish fed the diet devoid of thiamin developed neurological disorders, anorexia, reduced growth, and feed efficiency and increased mortality (Experiment 2 only) within 4–6 and 8–10 wk for Experiments 2 and 1, respectively. Low red blood cell count (RBC) and hematocrit (Ht) were observed in fish fed the thiamin‐deficient diet. Serum pyruvate was elevated in fish fed the thiamin unsupplemented diet. Serum lactate was not affected by dietary thiamin levels. Whole body protein was unaffected by dietary levels of thiamin. Body moisture and ash increased whereas body lipid decreased in fish fed the thiamin unsupplemented diets. None of these abnormalities were observed in fish fed the thiamin‐supplemented diets. Using the response curves determined by PROC NLMIXED to estimate dietary thiamin levels required for various response variables, a dietary thiamin level of 3.5 mg/kg diet was adequate for optimum growth, feed intake and efficiency, survival, prevention of neurological symptoms, and maintaining normal levels of RBC, Ht, serum pyruvate, and proximate body composition.     

Effects of dietary supplementation of a purified nucleotide mixture on immune function and disease and stress resistance in channel catfish,Ictalurus punctatus

Juvenile channel catfish (14.4 g average initial weight) were fed diets supplemented with a purified nucleotide mixture for 8 weeks. The mixture consisted of five nucleotides supplied on an equal basis as disodium salts at combined concentrations of 0 (control), 0.1%, 0.3%, 0.9% or 2.7% of diet. Addition of nucleotides to diet produced a dose‐dependent reduction in survival of channel catfish to Edwardsiella ictaluri. Although the reasons are unclear, the high levels of nucleotides supplemented in this study may have contributed to the decrease in disease resistance, and this decrease appeared to be unrelated to the innate immune (unaffected) and specific antibody (enhanced) responses. Stress resistance increased with a corresponding amelioration of the immunosuppressive effects of the stress response on non‐specific immunity (lysozyme and bactericidal activity) as nucleotides increased in diet. Use of exogenous nucleotides as a prophylactic treatment before culture‐related stress exposure may prove beneficial by decreasing the immunosuppressive effects of stress but not in prevention of ESC.     

Apoptosis in Ichthyophthirius multifiliis is associated with expression of the Fas receptor of theronts

The expression of type I membrane Fas receptors on the surface of Ichthyophthirius multifiliis (Ich) theronts and the possible association between Fas expression and theront apoptosis induced by the immune antibody was examined. Fas receptors were detected on the theront surface using fluorescein isothiocyanate-conjugated mouse monoclonal antibody against Fas. Fas-positive theronts significantly increased with time during in vitro incubation and with increasing theront concentration. Furthermore, the immune cutaneous antibody induced theront apoptosis; however, Fas ligand did not. A highly significant correlation was noted between theront Fas expression and immune cutaneous antibody-induced theront apoptosis. Numbers of apoptotic theronts increased with increasing number of Fas-positive theronts. The data indicated that theront apoptosis induced by immune cutaneous antibody appears to be positively correlated with the expression of Fas on the surface of Ich theronts.     

Metastatic mast cell tumour in a dog presenting with spinal pain

The clinical, advanced imaging and surgical features of a case of canine extradural mast cell tumour are presented. This rare location has only been reported once before as a primary lesion, although the present case may be a metastatic lesion. Given the frequent occurrence and treatment of cutaneous mast cell tumours in dogs, the finding of spinal metastasis is an important reminder to consider metastatic neoplasia in cases of previously treated cancers presenting with spinal pain.