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Summary

Effect of prior porcine respiratory coronavirus (PRCV) infection on replication of H1N1-influenza virus in the respiratory tract of swine was studied. In an initial experiment, 3 groups of 5 feeder pigs were studied. Pigs of 2 groups were inoculated sequentially with PRCV, followed by H1N1-influenza virus at 2- and 3-day intervals. Pigs of the other group were inoculated with H1N1-influenza virus only. Pigs were monitored clinically and examined for nasal excretion of influenza virus. In the singly influenza virus-inoculated group, 83% of nasal swab specimens were influenza virus-positive over a period of 6 days after inoculation. In the dually virus-inoculated groups, only 27% (2-day interval) and 53% (3-day interval) of nasal swab specimens were virus-positive over the same postinoculation period. However, clinical signs of infection in these dually inoculated pigs were more severe than those in the singly influenza virus-inoculated pigs. There were no significant differences in antibody responses against influenza virus among the 3 groups of pigs.

In a second experiment, 2 groups of pigs were studied. One group of pigs was inoculated sequentially with PRCV, followed by H1N1-influenza virus 2 days later; the other group was inoculated with H1N1-influenza virus only. Pigs of both groups were serially euthanatized on postinoculation days (pid) 1, 2, 3, and 4 (after influenza virus). At necropsy, influenza virus titer and immunofluorescence in lung tissue were determined and gross lung lesions were recorded. Influenza virus titer in the dually inoculated pigs (pid 1 and 2) was at least 100-fold reduced, compared with that in the corresponding singly inocu lated pigs, and fluorescence was either not detected (pid 1) or was scant (pid 2). Differences in influenza virus replication between pigs of dually and singly inoculated groups became gradually less pronounced at pid 3 and 4. Lung lesions in the dually virus-inoculated pigs were distinctly more severe than those in the corresponding singly virus-inoculated pigs, and became progressively more pronounced as time after influenza virus inoculation progressed.

These results indicate that PRCV infection may induce factors in the lungs that markedly interfere with replication and virus production during a subsequent influenza virus infection. On the other hand, clinical signs of infection and lung lesions were enhanced in the dually virus-inoculated pigs. It is believed that early nonspecific defense mechanisms in the lungs may have a role in the host antiviral response, as well as in development of lesions.

Free access
in American Journal of Veterinary Research

Summary

Adherence of Mycoplasma hyopneumoniae to the mucosa of the distal portion of the respiratory tract of swine is an important initial event in development of mycoplasmal pneumonia. A suitable in vitro model of adherence would be useful for investigation of mycoplasmal and host cell factors involved in this process. We have developed an adherence assay, using suspensions of porcine respiratory tract ciliated epithelial cells and M hyopneumoniae. Tracheal epithelial cells, collected by use of cytologic brushes, were mixed with broth cultures of M hyopneumoniae and the mixtures were incubated, diluted, vortexed, and sedimented. Pellets were spread on glass slides, stained with a fluorescent antibody against M hyopneumoniae, and evaluated by fluorescent microscopy. Fluorescence was observed principally among cilia on the ciliated tufts of epithelial cells. Only a few organisms were observed adhering on the nonciliated parts of ciliated cells or on other cell types. When mycoplasmas were preincubated with low dilutions of serum from swine convalescing from M hyopneumoniae disease, attachment was partially inhibited (P < 0.05). Significant inhibition of attachment was not observed when organisms were preincubated with higher dilutions of convalescent serum, with purified IgG from hyperimmune serum against M hyopneumoniae, or with low dilutions of lung lavage fluids (from convalescent swine) that contained specific IgA antibodies against M hyopneumoniae. Preincubation of the organisms with periodate and trypsin abolished attachment and formaldehyde decreased it (P < 0.05), whereas a variety of carbohydrates had no effect on attachment. Preincubation with dextran sulfate, ammonium sulfate, magnesium sulfate, and methionine reduced attachment (P < 0.05). Treatment of cell-Mycoplasma mixtures with the hydrophobic bond-breaking agent tetramethylurea, or incubation in absence of salt, or at low temperature also reduced attachment (P < 0.05). Attachment was not observed when ovine, rabbit, or guinea pig ciliated respiratory tract cells were mixed with M hyopneumoniae. Attachment of M dispar to porcine ciliated cells could not be detected, whereas M hyorhinis attached nonspecifically to all cell types in suspensions of porcine tracheal mucosa. These results indicate that adherence of M hyopneumoniae may be a host-specific event that is mediated by proteins and carbohydrates on the surface of the organism and by sulfur-containing molecules in the host cell membrane. The highly polarized location of the mycoplasmas on the cilia of epithelial cells indicates possible existence of stereospecific interactions between mycoplasmal adhesin(s) and receptor(s) on host cells. However, decreased adherence obtained by incubating Mycoplasma-ciliated cell mixtures with tetramethyl-urea, by incubating mixtures in absence of salt or at low temperature, indicates that nonspecific hydrophobic interactions might have a role in the attachment process.

Free access
in American Journal of Veterinary Research

contemporary CIV was isolated that same year. In mid February 2007, there was an outbreak of respiratory disease in dogs at a humane shelter. The outbreak involved all 27 dogs housed at the shelter at the time. Clinical signs included fever, lethargy, coughing

Full access
in American Journal of Veterinary Research

Abstract

Objective

To determine the apparent molecular weight for 24 ruminant respiratory syncytial viruses (RSV) on the basis of differences in the electrophoretic mobility of the phosphoprotein (P protein).

Procedure

29 bovine RSV (BRSV), 20 of which were not previously tested, 3 ovine RSV, and 1 caprine RSV isolates were selected for determination of electrophoretic mobility of the P protein. Virus radiolabeled with [35S]methionine was immunoprecipitated with polyclonal antiserum to BRSV and analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

Results

On the basis of apparent molecular size of the P protein, all isolates could be categorized into 2 electropherotypes, low molecular size of 36 kd and high molecular size of 38 kd. Twenty-three BRSV, the 3 ovine RSV, and 1 caprine RSV isolates had a high molecular size P protein; 6 BRSV isolates had a low molecular size P protein.

Conclusions

The apparent molecular size of the P protein of the ruminant RSV strains is greater than that of the human RSV subgroups, providing further evidence of their distinctiveness. Whether categorization of electrophoretic mobility of the P protein of BRSV underlies distinct antigenic subgroups, as it does in human RSV, requires further antigenic and genetic analysis.

Clinicai Relevance

Antigenic subgroups of ruminant RSV may have relevance in the development of new vaccines for control of the disease. (Am J Vet Res 1997;58:478–481)

Free access
in American Journal of Veterinary Research

Abstract

Objectives

To follow incidence of Pasteurella haemolytica (PH) in the upper respiratory tract of healthy calves at the farm and through the marketing process, and to determine the effect of vaccination on PH colonization of the upper respiratory tract and on the incidence of respiratory tract disease (RTD).

Animals

2- to 5-month-old calves (n = 104) from 4 farms.

Procedure

Calves were vaccinated with a killed PH serotype-1 product. Nasal secretion and tonsil wash specimens were cultured for PH, and serum antibody was measured by indirect hemagglutination. Calves with RTD were treated with tilmicosin phosphate.

Results

At the feedyard, 73 calves had RTD. The incidence of RTD was significantly related to the farm of origin, and was inversely related to the PH serum titer at the farm, but was not influenced by vaccination. Isolations of PH serotype 1 however, were reduced by vaccination. The major serotypes of PH encountered were 1 and 6.

Conclusion

Vaccination can reduce the frequency of colonization of the uoper respiratory tract by PH. (Am J Vet Res 1936;57:1317-1320)

Free access
in American Journal of Veterinary Research

Abstract

Objective

To determine the pathogenic potential of an adenovirus isolated from a goat.

Animals

14 colostrum-deprived, isolation-reared goat kids approximately 3 weeks old.

Procedure

Kids were inoculated with either cell culture fluid containing adenovirus (n = 10) or uninfected cell culture fluid (n = 4): 2 ml transtracheally and 1 ml/nostril. Clinical signs of disease and rectal temperature were recorded daily; nasal secretion and fecal specimens were collected daily. Control kids were necropsied, 2/d, on postinoculation days (PID) 5 and 10. Virus-inoculated kids were necropsied on PID 3, 5, 7, 10, and 28. After necropsy, lung, liver, kidney, and brain specimens were aseptically collected for virus isolation attempts. Tracheal fluid was collected on sterile cotton swabs. Turbinate, trachea, lung, mediastinal lymph node, liver, kidney, duodenum, jejunum, ileum, mesenteric lymph node, colon, and brain specimens were collected for histologic evaluation.

Results

Kids developed mild-to-moderate clinical respiratory tract infection. Virus was recovered consistently from nasal secretion and sporadically from fecal specimens. Grossly, there were multiple areas of atelectasis and hyperemia, principally in the cranioventral portion of the lungs. Microscopically, there was detachment and sloughing of foci of epithelial cells of the terminal bronchioles and alveoli. In kids necropsied late in the disease, these changes were accompanied by hyperplasia of type-II epithelial cells. Viral inclusions were not an obvious feature, but a few cells contained probable inclusions.

Conclusions and Clinical Relevance

The caprine adenovirus reported here is capable of inducing respiratory tract disease and lesions in the lungs of young kids. (Am J Vet Res 1997;58:608–611)

Free access
in American Journal of Veterinary Research

Abstract

Objective

To determine whether porcine reproductive and respiratory syndrome virus plaque variants vary in their pathogenicity in causing late-term reproductive failure.

Design

Four groups of 2 sows each at 86 days of gestation were inoculated intranasally with PRRSV small (MN-Hs) and large (MN-HI) plaque variants, field isolate, and cell culture medium, respectively. In addition, 2 sows each at 86 days of gestation were inoculated intranasally or IM with MN-Hs virus.

Animals

14 pregnant sows.

Procedure

Inoculated sows were allowed to deliver at term, and each litter was examined for clinical abnormality and presence of virus.

Results

Two sows infected with MN-Hs virus delivered 14 live and 5 dead pigs, whereas 2 sows infected with MN-HI virus delivered 0 live and 25 dead pigs. Two sows inoculated with a field isolate (MN-W) delivered 10 live and 20 dead pigs. Two control sows had 26 normal fetuses at slaughter at 107 days of gestation. Virus was isolated from 16 (66.7%) of 24 liveborn pigs, 9 (64.3%) of 14 stillborn pigs, and 3 (12.0%) of 25 mummified fetuses of the 6 infected sows. Subsequently, 4 MN-Hs-infected sows delivered 40 live and 11 dead pigs.

Conclusions

Marked difference in the pathogenicity in pregnant sows between porcine reproductive and respiratory syndrome virus strains was documented. The MN-Hs virus is considered to be of low pathogenicity, but the other viruses are highly pathogenic for late-term pregnant sows.(Am J Vet Res 1996;57:320-323)

Free access
in American Journal of Veterinary Research

Summary

Despite the high incidence of distal respiratory tract infection of undetermined cause on farms, to our knowledge, the microbiologic effects of conventional antimicrobial treatment for this condition have not been studied. We evaluated the possible pathogenic role of bacterial isolates from the distal airways of foals with clinical respiratory tract disease, by correlating changes in their numbers (increase or decrease) with clinical, endoscopic, and pulmonary cytologic signs of disease resolution during treatment with antimicrobial drugs. We also determined qualitative changes in in vitro antimicrobial susceptibility of bacterial isolates after 7 days of treatment and relapse rate of foals. Significant (P < 0.05) decrease in the numbers of an isolate in the airways was considered strong evidence of a pathogenic role in this disease syndrome. Foals with endoscopically confirmed distal respiratory tract infection (drti; n = 65) were selected at random for treatment (n = 56) or nontreatment (n = 9), and bronchial lavage specimens were cultured and evaluated cytologically before and after 7 days of treatment with trimethoprim-sulfamethoxazole (tms) and a β-lactam drug (penicillin, ampicillin, or sulbactam-ampicillin), the standard treatment in all foals. The effect of treatment was to abruptly reduce the clinical (nasal discharge, cough, adventitious lung sounds) and cytologic signs of airway infection. Severity of disease in nontreated foals, however, did not change or did worsen over time. Reduction in the frequency and numbers of Streptococcus zooepidemicus isolated during treatment supported a causal role for this organism in the clinical syndrome observed. On the other hand, the frequency of non-Str zooepidemicus isolates (eg, Staphylococcus epidermidis, Streptomyces spp, α-hemolytic streptococci) actually increased during treatment, compatible with a commensal or competitive role for these organisms. Significantly (P < 0.001) more pretreatment isolates were susceptible in vitro to either tms or β-lactam drugs than to β-lactam drugs alone; more posttreatment isolates were susceptible to either tms or β-lactam than to either drug alone. These data indicate that there may be some benefit to combined use of tms plus β-lactam drugs in foals with drti. Mean ± sem relapse rate was 31 ± 6% (range 0 to 57%); risk factors (clinical signs of disease, laboratory variables) for relapse could not be identified. In conclusion, treatment resulted in significant (P < 0.001) reduction in airway inflammation in foals with clinical drti. The high reinfection rate indicates that a predisposing factor, possibly age-related immunodeficiency, may predispose foals to illness and persists after treatment.

Free access
in American Journal of Veterinary Research

Summary

Pregnant gilts were exposed to porcine reproductive and respiratory syndrome virus (prrsv ) by iv inoculation at or about gestation day 30 (3 gilts), 50 (3 gilts), 70 (3 gilts), or 90 (5 gilts) to investigate the likelihood of transplacental infection with prrsvat various stages of gestation. At or about 3, 6, and 9 weeks after exposure, gilts were either euthanatized while still pregnant or allowed to farrow. Gilts and pigs were observed for clinical signs of infection, and gilts, pigs, and fetuses were tested for prrsvand homologous antibody. All gilts were healthy throughout the study, except that farrowing was sometimes difficult and prolonged, and 2 gilts failed to farrow the entire litter. One gilt farrowed on day 111 of gestation; all others farrowed on day 114 or later. Porcine reproductive and respiratory virus was isolated from significantly (x2 test, P < 0.01) more fetuses and live and stillborn pigs of the 5 gilts that were infected at 90 or 92 days of gestation than from the fetuses and live and stillborn pigs of the 9 gilts that were infected at 72 or fewer days of gestation (ie, 33 of 44, 75% vs 3 of 78,4%). After initial infection, prrsvwas isolated from gilts and their pigs for a maximum of 3 weeks and 8 to 11 weeks, respectively. Findings of this study, with regard to the temporal aspects of transplacental infection, may help explain why natural epizootics of prrsv-induced maternal reproductive failure are often recognized principally as problems of late-term gestation and neonatal survival.

Free access
in American Journal of Veterinary Research

SUMMARY

The ability of 25 Pasteurella multocida isolates to adhere in vitro to porcine respiratory tract mucus was examined. Microplate wells were coated with crude mucus preparation, then bacteria were added. After incubation, unbound bacteria were removed by washing, and the number of mucus-bound bacteria was estimated by quantitation of the adherent colony-forming units and by use of an elisa. Pasteurella multocida had affinity to respiratory tract mucus, although significant differences were not observed in affinity of capsular type-A and type-D isolates. Preliminary characterization, using ultrafiltration, gel filtration chromatography, electrophoresis, and enzymatic treatments, indicated that the receptors may be a class of protein molecules of low molecular weight (< 25,000). The origin of these receptors, however, is not known at this time.

Free access
in American Journal of Veterinary Research