Evaluation of the ability of tilmicosin to prevent adherence of Mycoplasma hyopneumoniae to cilia using a differentiated swine respiratory epithelial culture system.

Mycoplasmal pneumonia caused by Mycoplasma hyopneumoniae is a serious economic problem for swine producers in the United States. Mycoplasma hyopneumoniae colonizes ciliated respiratory epithelial cells. The organism has been shown to be sensitive to tilmicosin, a synthetic macrolide, in antibiotic sensitivity assays. The efficacy of tilmicosin to inhibit adherence of M. hyopneumoniae to ciliated epithelial cells without direct contact between the antibiotic and the organism was evaluated using in vitro methods. The study demonstrated that tilmicosin inhibited the adherence of M. hyopneumoniae at the highest level tested in the system (2 microg/ml) suggesting that tilmicosin may be efficacious against mycoplasmal pneumonia.

Field isolates of Mycoplasma ovipneumoniae exhibit distinct cytopathic effects in ovine tracheal organ cultures.

Ovine tracheal ring explants were infected with four different Mycoplasma ovipneumoniae and one M. arginini field isolate and their ability to induce cytopathic effects was tested by measuring ciliary activity and intracellular calmodulin release. Infected tracheal rings showed significantly decreased ciliary activity as compared to the non-infected control rings. There were, however, marked differences between isolates in the onset and severity of the effects which correlated with their ability to produce hydrogen peroxide. Infected tracheal rings released more calmodulin than the non-infected controls. The amount of calmodulin released also varied between isolates, and somewhat reflected the degree of loss of ciliary activity in the corresponding rings induced by the different isolates. Light and electron microscopic examinations of infected tracheal rings revealed disorganisation and sloughing of the epithelium, and association of mycoplasmas only with the cilia. Following repeated in vitro passages, the organisms had reduced ability to inhibit ciliary activity which correlated with decreased hydrogen peroxide production. Addition of catalase to the organ cultures delayed loss of ciliary activity. These results suggest that M. ovipneumoniae induced ciliostasis in ovine tracheal ring explants which correlated with hydrogen peroxide production. Furthermore, these M. ovipneumoniae-induced injuries to respiratory epithelial cells could contribute to the role that this organism may play in sheep respiratory disease.

Effect of environmental variables in turkey confinement houses on airborne Aspergillus and mycoflora composition.

Environmental conditions and airborne mycoflora were measured concurrently in 10 turkey confinement houses during warm and cold weather. The following variables in the environment were measured: numbers of feed- and litter-associated yeast and mold fungi, temperature, relative humidity, airspeed, carbon dioxide and ammonia concentration, airborne bacteria, and airborne particulate mass, particle number, and particle size distribution. Winter air in turkey confinement houses contained significantly higher concentrations of Aspergillus, Scopulariopsis, and Mucor sp. and significantly lower concentrations of Cladosporium, Fusarium, and Alternaria sp. when compared with summer air. Significantly greater numbers of Mucor sp. were recovered per cubic meter of air where the current turkey flock was present less than 100 d when compared to houses where the current flock resided 100 d or more. Management decisions regarding control of the internal environment of turkey confinement houses apparently influence airborne mycoflora composition.

Production of gliotoxin during the pathogenic state in turkey poults by Aspergillus fumigatus Fresenius.

Turkey poults were given either of two different dosages of two different gliotoxin-producing strains of Aspergillus fumigatus. Infected lung tissue was examined postmortem for the presence of gliotoxin. Gliotoxin was found in lung tissue of ten poults infected with one strain and in seven of ten poults infected with the other strain. Concentrations of gliotoxin in the tissue exceeded 6 ppm in some of the infected tissues. The concentration of gliotoxin found in infected tissue did not appear to be correlated with the dosage of organism given. Considering the pathologic changes observed in turkey poults with aspergillosis and the production of gliotoxin during the pathogenic state in turkey poults, gliotoxin is considered likely to be involved in avian aspergillosis.

Ciliostasis and loss of cilia induced by Mycoplasma hyopneumoniae in porcine tracheal organ cultures.

In vivo- and in vitro-grown Mycoplasma hyopneumoniae organisms were inoculated onto newborn piglet tracheal organ cultures to provide a model for interaction of this organism with ciliated respiratory epithelium. Ciliostasis and loss of cilia in tracheal rings were induced by M. hyopneumoniae grown in vivo and with low-passage cultures when grown in vitro. Levels of calmodulin or dehydrogenase enzymes in tracheal ring epithelium were not altered even though ciliostasis and loss of cilia induced by M. hyopneumoniae were extensive. The capacity for inducing epithelial damage diminished with in vitro passage of the organism. Attempts to induce higher-passage cultures to attach to cilia, cause ciliostasis, or cause ciliary damage by supplementation of mycoplasmal medium with porcine lung extract failed. Epithelial damage induced by M. hyopneumoniae in tracheal rings was averted by using porcine immune serum or by separating the organisms from ciliated epithelium with a 0.1-microns-pore-size membrane. Attachment, or at least close association, of M. hyopneumoniae to ciliated epithelium appeared to be necessary to induce ciliostasis and loss of cilia in this model.

Aspergillus fumigatus keratitis with intraocular invasion in 15-day-old chicks.

Eye infections were initially observed in single-comb white leghorn breeder chicks at 5 days of age, and morbidity increased from 0.05% to 1.5% after debeaking at 7 days of age. All chicks necropsied at 15 days of age had cheesy yellow exudate within the conjunctival sac of one eye and small (1 mm diameter) white nodular lesions in lungs and on thoracic air-sac membranes. Histopathologic examination of the eyes revealed septate fungal hyphae and inflammatory cells in the anterior chamber, cornea, and conjunctival sac. Similar fungal hyphae were present within lung granulomas. Aspergillus fumigatus was isolated from the eyes. Eye infections were the only health problem reported for several consecutive flocks on this farm. Elimination of moldy feed from the diet and environment and proper management of sawdust litter have prevented fungal ophthalmitis in subsequent flocks.

Effect of building ventilation design on environment and performance of turkeys.

Environmental variables in 10 commercial turkey confinement buildings, representing 2 natural ventilation designs, were measured during summer and the following winter. Sliding doors spaced at intervals along the walls of 5 of the buildings provided about 35% opening, and continuous wall curtains provided 60 to 80% opening in the other 5 buildings. Environmental variables assessed included airspeed; temperature; relative humidity; gases; particle number, size, and mass per cubic meter of air; and colonies of bacteria, yeasts, and other fungi per cubic meter of air. Colonies of yeasts and other fungi were quantitated in feed and litter. For most of the variables evaluated, significant differences were not attributable to building ventilation design; however, in winter, the total mass of particulate matter per cubic meter of air was higher in the curtain-type houses, compared with sliding door-type houses. Ammonia concentration in the air of sliding door-type houses progressively increased during summer and winter sampling periods. A significant effect of building ventilation design on turkey performance was not detected when using mortality, average daily gain, feed conversion, condemnations at slaughter, or average individual bird weight as measures of production.

Enhancement of the increase in intracellular calcium concentration in stimulated neutrophils by Mycoplasma hyopneumoniae.

Neutrophils isolated from the peripheral blood of pigs free of infection with Mycoplasma hyopneumoniae were loaded with a fluorescent indicator (Fura-2) for detection of cytosolic free calcium concentration. The kinetics of the intracellular calcium flux were examined after incubation with or without a pathogenic or a non-pathogenic strain of M. hyopneumoniae. The basal intracellular calcium concentration was not altered by incubation with M. hyopneumoniae. However, the relative increase in cytoplasmic calcium concentration caused by the addition of opsonized zymosan was significantly (p < 0.05) higher in neutrophils incubated with M. hyopneumoniae as compared to neutrophils not incubated with M. hyopneumoniae. Additionally, after zymosan stimulation, the intracellular calcium concentration was greater in neutrophils incubated with a pathogenic strain of M. hyopneumoniae than in those incubated with a non-pathogenic strain. This suggests that M. hyopneumoniae alters the signal transduction mechanisms in neutrophils and that this alteration may be related to virulence.

Histochemical and morphologic changes of porcine airway epithelial cells in response to infection with Mycoplasma hyopneumoniae.

Mycoplasma hyopneumoniae causes pneumonia in pigs. The effect of infection by this organism on histochemical characteristics of airway mucin within epithelial cells was studied. Seven- to 10-week-old pigs were inoculated intratracheally with M hyopneumoniae or culture broth, and lung tissues were collected from inoculated and control pigs at 2, 4, and 6 weeks after inoculation. Tissue sections were stained with periodic acid-Schiff/Alcian blue, pH 2.5 or high iron diamine/Alcian blue. Histologic features of randomly selected bronchi, bronchioles, and submucosal glands were compared in sections stained with periodic acid-Schiff/Alcian blue. Bronchial goblet cell sulfomucin and sialomucin were quantitated by image analysis of sections stained with high iron diamine/Alcian blue. Bronchi and bronchioles of infected pigs contained proportionately fewer goblet cells with mucin at all stages of infection than age-matched control pigs. Goblet cells in bronchi of infected pigs contained significantly less total mucin and sialomucin, and significantly more sulfomucin than goblet cells of control pigs. Increased sulfated mucin in bronchial goblet cells may reflect altered glycoprotein production or secretion in response to infection with M hyopneumoniae.

Rhodococcus equi in fecal and environmental samples from Kansas horse farms.

Horse farms in Kansas were surveyed for the incidence of Rhodococcus equi. Fecal specimens and soil or cobweb samples were collected from each farm and cultured on selective media. One control farm (with no history of R. equi infection), one farm which had an outbreak 3 and 4 years previously and 2 farms which had R. equi-infected foals that season were surveyed. In addition, fecal samples from 21 horses hospitalized in the Kansas State University Veterinary Hospital were cultured. There was no significant difference in the incidence of R. equi in fecal samples from the 2 farms with recent disease problems. The farm with a history of disease had a significantly higher percentage of positive fecal cultures than the 2 farms with a more recent history of disease. Neither ration composition nor sex of the horses appeared to affect the fecal culture results. Fecal samples from 2 birds on 1 farm were positive for R. equi.