Evaluation of a monoclonal blocking ELISA and IHA for antibodies to Mycoplasma hyopneumoniae in SPF-pig herds.

A monoclonal blocking enzyme-linked immunosorbent assay (ELISA) and an indirect haemagglutination assay (IHA) were applied to serum samples from 124 specific pathogen-free (SPF) breeding and multiplying herds, which participate in the routine serological surveillance of the Danish SPF programme. Clinical and pathological observations of the herds and microbiological culturing of Mycoplasma hyopneumoniae were used to calculate herd sensitivity, herd specificity and herd predictive values for the two serological assays. The ELISA was superior to the IHA in herd sensitivity and herd specificity, with values of 93 per cent and 96 per cent, respectively, for the ELISA, and 61 per cent and 92 per cent for the IHA. During the six month period of evaluation 2.5 per cent of the herds were infected with M hyopneumoniae each month. At this level the IHA was found to have a positive herd predictive value of 16 per cent, compared with 39 per cent for the ELISA. The negative herd-predictive value on the same level was 99.8 per cent for the ELISA and 98.9 per cent for the IHA. If the assays were applied to a group of herds with a herd prevalence of M hyopneumoniae infection of 30 per cent (as is the case with the production herds in the Danish SPF programme) the predictive value of a positive herd diagnosis would be 91 per cent for the ELISA and 76 per cent for the IHA, and the predictive value of a negative herd diagnosis would be 97 per cent with the ELISA and 85 per cent with the IHA.(ABSTRACT TRUNCATED AT 250 WORDS)     

MAGNETIC RESONANCE, ULTRASOUND AND HISTOPATHOLOGIC CORRELATION OF ACUTE AND HEALING EQUINE TENDON INJURIES

Ultrasonography and MRI have become valuable tools for imaging of tendon injuries. The current study examines the histopathologic basis for the imaging abnormalities. Five injured equine forelimbs and two normal contralateral limbs were studied with high resolution real time ultrasound and MRI. Histologic sections were made and correlated with the diagnostic images. All lesions were readily seen by both modalities. Lesions characterized by hemorrhage, edma, and cellular infiltration were sonolucent on ultrasound and bright on MRI images. MRI returned to normal as fibrogenesis ensued. Ultrasound images remained abnormal until fibrillar reallgnment occured with completion of the healing process. High resolution real time ultrasound and MRI both accurately reflect the tissue abnormalities in acute tendon injuries. The injured horse does appear to be an effective model for correlative imaging studies of tendon injuries.     

Arthropathy associated with Brucella abortus strain 19 vaccination in cattle. II. Experimental studies

Attempts to reproduce in calves the arthropathy associated with Brucella abortus strain 19 (S19) vaccination by direct intra-articular injection of S19 or virulent Brucella cells were unsuccessful. Intra-articular injection of rabbits and a calf with immune complexes isolated from the synovial fluid of a field case produced clinical and histological signs of arthropathy accompanied by the development of rheumatoid factor.     

Chylothorax associated with cryptococcal mediastinal granuloma in a cat

A 10-year-old neutered female cat had chylothorax, precaval syndrome, and a mediastinal granuloma resulting from infection with Cryptococcus neoformans. Diagnosis of a chylous effusion was made by cytologic examination of pleural fluid and by finding higher triglyceride levels in the effusion than in serum (825 vs. 64 mg/dl, respectively). Postmortem examination revealed cryptococcal organisms in the mediastinal granuloma, lungs, cerebral meninges, and connective tissues adjacent to the thyroid gland. Chylous effusion in a cat associated with cryptococcosis has not been reported previously. Cryptococcosis should be included in the differential diagnosis in chylous effusions in cats.     

Bilateral phacofragmentation in a New Zealand fur seal (Arctocephalus forsteri).

Ophthalmic examination under anesthesia of a 5-yr-old, captive, male New Zealand fur seal (Arctocephalus forsteri) with reduced visual capacity revealed bilateral cataracts. Surgical extraction of the lenses was performed by phacofragmentation. Preoperative mydriasis was unsuccessful using common topical mydriatic agents. Intraoperative intracameral epinephrine 1:10,000 produced minimal pupillary dilation, but excellent dilation was achieved with intracameral epinephrine at 1:1,000. Postoperative uveitis was controlled with oral prednisolone. Postoperative complications included generalized corneal edema persisting for approximately 1 wk and corneal ulceration caused by the lateral canthotomy sutures. Vision was restored in both eyes.     

Expression of the tobacco β-1,3-glucanase gene, PR-2d, following induction of SAR with Peronospora tabacina

Systemic acquired resistance (SAR) is induced following inoculation of Peronospora tabacina sporangia into the stems of Nicotiana tabacum plants highly susceptible to the pathogen. Previous results have shown that accumulation of acidic β-1,3-glucanases (PR-2's) following induction of SAR by P. tabacina may contribute to resistance to P. tabacina. We showed that up-regulation of the PR-2 gene, PR-2d, following stem inoculation with P. tabacina, is associated with SAR. Studies using plants transformed with GUS constructs containing the full length promoter from PR-2d or promoter deletions, provided evidence that a previously characterized regulatory element that is involved in response to salicylic acid (SA), may be involved in regulation of PR-2d following induction of SAR with P. tabacina. This work provides evidence that regulation of PR-2 genes during P. tabacina-induced SAR may be similar to regulation of these genes during infection of N-gene tobacco by TMV or following exogenous application of SA, and provides further support for the role of SA in regulation of genes during P. tabacina-induced SAR.     

Characterization and PCR-based Typing of Xanthomonas campestris pv. vesicatoria from Peppers and Tomatoes in Serbia

During the last two decades bacterial strains associated with necrotic leaf spots of pepper and tomato fruit spots were collected in Serbia. Twenty-eight strains isolated from pepper and six from tomato were characterized. A study of their physiological and pathological characteristics, and fatty acid composition analysis revealed that all of the strains belong to Xanthomonas campestris pv. vesicatoria. Being non-amylolytic and non-pectolytic, pathogenic on pepper but not on tomato, containing lower amounts of fatty acid 15 : 0 ante–iso, the pepper strains were designated as members of the A group of X. campestris pv. vesicatoria. However, the tomato strains hydrolyzed starch and pectate, caused compatible reactions on tomato but not on pepper, had higher percent of 15 : 0 ante–iso fatty acid, and were classified into B phenotypic group and identified as X. vesicatoria. PCR primers were developed which amplified conserved DNA regions related to the hrp genes of different strains of X. campestris pv. vesicatoria associated with pepper and tomato. Restriction analysis of the PCR product resulted in different patterns and enabled grouping of the strains into four groups. When xanthomonads isolated from pepper and tomato in Serbia were analyzed, they clustered into two groups corresponding to the grouping based on their physiological and pathological characteristics. According to the reaction of pepper and tomato differential varieties, the strains from pepper belong to races P7 and P8 and tomato strains belong to the race T2. All strains were sensitive to copper and streptomycin. Advantages and disadvantages of various bacterial spot management practices are discussed.     

Effect of hay type on cecal and fecal microbiome and fermentation parameters in horses

The effect of hay type on the microbiome of the equine gastrointestinal tract is relatively unexplored. Our objective was to characterize the cecal and fecal microbiome of mature horses consuming alfalfa or Smooth Bromegrass (brome) hay. Six cecally cannulated horses were used in a split-plot design run as a crossover in two periods. The whole plot treatment was ad libitum access to brome or alfalfa hay fed over two 21-d acclimation periods with subplots of sampling location (cecum and rectum) and sampling hour. Each acclimation period was followed by a 24-h collection period where cecal and fecal samples were collected every 3 h for analysis of pH and volatile fatty acids (VFA). Fecal and cecal samples were pooled and sent to a commercial lab (MR DNA, Shallowater, TX) for the amplification of the V4 region of the 16S rRNA gene and sequenced using Illumina HiSeq. The main effects of hay on VFA, pH, and taxonomic abundances were analyzed using the MIXED procedure of SAS 9.4 with fixed effects of hay, hour, location, period, and all possible interactions and random effect of horse. Alpha and beta diversities were analyzed using the R Dame package. Horses fed alfalfa had greater fecal than cecal pH (P ≤ 0.05), whereas horses fed brome had greater cecal than fecal pH (P ≤ 0.05). Regardless of hay type, total VFA concentrations were greater (P ≤ 0.05) in the cecum than in feces, and alfalfa resulted in greater (P ≤ 0.05) VFA concentrations than brome in both sampling locations. Alpha diversity was greater (P ≤ 0.05) in fecal compared with cecal samples. Microbial community structure within each sampling location and hay type differed from one another (P ≤ 0.05). Bacteroidetes were greater (P ≤ 0.05) in the cecum compared with the rectum, regardless of hay type. Firmicutes and Firmicutes:Bacteroidetes were greater (P ≤ 0.05) in the feces compared with cecal samples of alfalfa-fed horses. In all, fermentation parameters and bacterial abundances were impacted by hay type and sampling location in the hindgut.