Testing of a hyperimmune serum in calves infected intranasally with Mycoplasma bovis

Mycoplasma (M.) bovis hyperimmune serum was subcutaneously injected to 16 of 26 calves repeatedly intranasally infected with M. bovis, during and/or after experimental infection. Antibody titres between 1:32 and 1:256 were recorded by means of indirect haemagglutination from the calves treated. Transmission of film-inhibitory antibodies failed to work. Neither clinical manifestations nor pathologico-anatomic alterations to the lungs of experimentally infected animals were mitigated by hyperimmune serum treatment. M. bovis, in high germ counts, was re-isolated from nasal swabs, trachea, pulmonary lymph nodes, and inflammatory lung tissue of both treated and untreated calves.     

On the aetiopathogenesis of Mycoplasma pneumonia in calf

A pure culture of Mycoplasma (M.) bovis was isolated from calves with respiratory disease, exhibiting the picture of lymphohistiocytic proliferative pneumonia with presence of eosinophil plasmatic cells. A mixed infection of M. bovis and Pasteurella (P.) multocida was demonstrated in calves with exudative pneumonia. Both M. bovis and Haemophilus (H.) somnus were recovered from calves with necrotic pneumonia. All 3 organisms--M. bovis, P. multocida, and H. somnus--were present in cases of exudative-necrotic pneumonia. It was also shown that M. bovis played a primary role in the aetiopathogenesis of respiratory diseases caused by mixed infections.     

Pathological and immunological changes after challenge infection with Pasteurella multocida in naive and immunized calves

Challenge infections of calves with Pasteurella multocida were established to characterize the local inflammatory response and determine the effect of previous exposure to live bacteria on the post-challenge immune response. Experimental infections were established by intratracheal inoculation of P. multocida in both naive calves and calves that had been previously vaccinated with two subcutaneous (s.c.) injections of live bacteria. Histological, immunohistological and cytokine expression studies were performed on bronchoalveolar lavage (BAL) samples, lung parenchymal tissues and lung lymph nodes (LN). In comparison to uninfected control animals in which no lung lesions were observed, a patchy to confluent bronchopneumonia was observed following infection of naive calves, characterized by abscess formation, haemorrhage, oedema and suppurative consolidation. Cellular analysis following infection of naive animals was characterized by an influx of neutrophils in the BAL, with macrophages and dendritic cells observed in the lesion perimeter. A significant increase in the number of CD8(+) blasts expressing MHC (major histocompatibility) II was also observed in the BAL of infected calves. Decreased expression of interleukin (IL)-1 beta and increased expression of IL-8 compared to naive unchallenged controls was apparent in lung LN. In comparison, a more limited pathology was observed in vaccinated animals post-challenge, indicating partial protection conferred by the s.c. immunization with live bacteria. Studies of vaccinated animals showed the presence of bronchial-associated lymphoid tissue (BALT) in the lung tissue and an increase in the number of B-cells and CD4(+) T-cells expressing MHCII in the lung LN after challenge. In contrast to primary infection, there was no significant influx of neutrophils in the BAL. Instead, a population of newly recruited monocytes/macrophages was observed. Increased IL-2 expression and decreased IL-8 expression was observed in the LN, while IL-1 beta expression was not detected. The reduced neutrophil and increase monocyte response in the vaccinated calves may be associated with significant changes in the gamma delta T lymphocyte population in the BAL.     

Clinical study of the disease of calves associated with Mycoplasma bovis infection

Clinical, bacteriological and serological examination of 35 calves from the age of 5 to 26 days was performed in a Holstein-Friesian dairy herd endemically infected with Mycoplasma bovis. M. bovis was isolated from 48.6% of nasal swabs taken from the calves at the age of 5 days, and from 91.4% of the same calves at the age of 26 days, indicating the gradual spread of infection. The isolation rate of Pasteurella multocida did not change much, and varied from 28.6 to 25.7%. No P. haemolytica could be detected. In addition to M. bovis and P. multocida, the herd was also infected with different viruses (including bovine viral diarrhoea virus, infectious bovine rhinotracheitis virus, bovine adenoviruses, parainfluenza-3 virus, and bovine respiratory syncytial virus) as a large proportion of the sera of newborn calves contained colostral antibodies against these viruses. In most of the newborn calves severe clinical signs (fever, depression, inappetence, hyperventilation, dyspnoea, nasal discharge and coughing) due to M. bovis infection developed. The clinical signs appeared already on the fifth day of life, and their incidence was the highest at the age of 10 to 15 days. Three calves (8.6%) died as a result of severe serofibrinous pneumonia. The surviving calves showed very poor weight gain (ranging from 1.5 to 3.5 kg) during the first two weeks of life.     

Effect of a new macrolide antibiotic (tilmicosin) on pneumonia experimentally induced in calves by Mycoplasma bovis and Pasteurella haemolytica

Two gnotobiotic calves were treated once with tilmicosin (20 mg kg-1) six hours before they were infected by the intratracheal route with Mycoplasma bovis and Pasteurella haemolytica serotype 1. This treatment prevented colonisation of the lungs by P haemolytica and considerably reduced colonisation by M bovis, and the clinical scores and the extent of pneumonic consolidation, compared with two untreated gnotobiotic calves, both of which had to be killed in extremis for humanitarian reasons within 24 hours of infection. In a second experiment, 10 conventionally reared calves were similarly exposed to infection and, at the onset of clinical disease, five were treated once with tilmicosin (20 mg kg-1). Colonisation by P haemolytica and M bovis, the clinical scores and extent of pneumonic consolidation were suppressed or greatly reduced in the treated compared with the untreated calves, one of which had to be killed in extremis two days after infection. It was concluded that tilmicosin had a beneficial effect.     

The microbial flora of the respiratory tract in feedlot calves: associations between nasopharyngeal and bronchoalveolar lavage cultures.

The upper and lower respiratory tracts of 59 feedlot calves with clinical signs of naturally occurring respiratory disease (cases) and 60 comparison (control) animals were cultured before treatment, using nasopharyngeal swabs (NPS) and bronchoalveolar lavage (BAL). The most prevalent organisms were Pasteurella multocida and Mycoplasma bovis. Isolations of P. multocida from NPS and BAL fluid were found to be significantly associated with morbidity (p less than or equal to 0.05), but the frequency with which other organisms were isolated from the nasopharynx and lungs was similar in cases and controls. There was evidence of moderate agreement between NPS and BAL isolates at the individual calf level using the kappa statistic, (range of kappa values = 0.47-0.61) but the variability of the kappa statistics was large. Therefore, in an individual calf NPS cultures did not accurately predict BAL cultures. The NPS and BAL culture results were quite similar at the group level, however.     

Increased pulmonary secretion of tumor necrosis factor-alpha in calves experimentally infected with bovine respiratory syncytial virus

Bovine respiratory syncytial virus (BRSV) is an important cause of respiratory disease among calves in the Danish cattle industry. An experimental BRSV infection model was used to study the pathogenesis of the disease in calves. Broncho alveolar lung lavage (BAL) was performed on 28 Jersey calves, of which 23 were experimentally infected with BRSV and five were given a mock inoculum. The presence of the cytokine tumor necrosis factor alpha (TNF-alpha) in the BAL fluids was detected and quantified by a capture ELISA. TNF-alpha was detected in 21 of the infected animals. The amount of TNF-alpha in the BAL fluid of calves killed post inoculation day (PID) 2 and 4 was at the same very low level as in the uninfected control animals. Large amounts of TNF-alpha were detected on PID 6, maximum levels of TNF-alpha were reached on PID 7, and smaller amounts of TNF-alpha were seen on PID 8. The high levels of TNF-alpha appeared on the days where severe lung lesions and clinical signs were obvious and the amounts of BRSV-antigen were at their greatest. Although Pasteurellaceae were isolated from some of the BRSV-infected calves, calves treated with antibiotics before and through the whole period of the infection, as well as BRSV-infected calves free of bacteria reached the same level of TNF-alpha as animals from which bacteria were isolated from the lungs. It is concluded that significant quantities of TNF-alpha are produced in the lungs of the calves on PID 6-7 of BRSV infection. The involvement of TNF-alpha in the pathogenesis of, as well as the anti-viral immune response against, BRSV infection is discussed.     

The efficacy of valnemulin (Econor) in the control of disease caused by experimental infection of calves with Mycoplasma bovis

Mycoplasma bovis infection was experimentally induced in groups of six young calves. A further group was uninfected and served as a control. Ten days after infection, medication with either enrofloxacin (Baytril, Bayer) or valnemulin (Econor, Novartis) was instituted via the milk replacer for a further 10 days, after which all calves were killed. Infection resulted in depression, pyrexia, inappetance and prominent respiratory signs. Arthritis occurred in two animals and two (unmedicated) animals died. At post-mortem examination extensive lesions were present in the lungs and M. bovis was re-isolated from infected unmedicated calves' lungs. Medication with either enrofloxacin or valnemulin resulted in a rapid diminution of clinical signs, restoration of appetite and reversal of weight loss. Isolation of Pasteurella multocida from the calves' lungs was suppressed by both medicaments. Valnemulin resulted in a more rapid reduction of clinical scores and eliminated M. bovis from the lungs more effectively than enrofloxacin.     

Changes in the bacterial flora of the upper and lower respiratory tracts and bronchoalveolar lavage differential cell counts in feedlot calves treated for respiratory diseases.

Serial nasopharyngeal swab and bronchoalveolar lavage cultures were used to estimate changes in the bacterial flora of the respiratory tracts of calves during the first month after arrival in the feedlot. Bronchoalveolar lavage (BAL) differential cell counts served to evaluate pulmonary inflammatory changes during this period. Two groups of calves were studied, one consisting of clinically normal controls (n = 60), the other, of cases (n = 59) which received treatment for respiratory disease (penicillin +/- trimethoprimsulfadoxine). A variety of organisms, including Pasteurella multocida, Pasteurella haemolytica, Haemophilus somnus, Mycoplasma bovis and Mycoplasma bovirhinis, were present in the upper and lower airways of both groups during the postarrival period. With the exception of M. bovis, an overall decline in the prevalence of these organisms was observed during the course of the study. In cases, there was a marked decrease in the number of Pasteurella spp. and H. somnus isolates immediately following treatment. For the Pasteurella spp., however, this effect was shortlived as they often appeared to recolonize the respiratory tract within eight days of terminating antimicrobial therapy. Treatment did not appear to affect the frequency of isolating M. bovis. Its prevalence, in both groups of calves, increased to levels approaching 100% during the course of the study. All Pasteurella spp. isolates were tested for susceptibility to several commonly used antimicrobials. Resistance was only evident among P. haemolytica isolated from cases and in every instance this was to a combination of penicillin, ampicillin and tetracycline. Significantly more isolates were resistant after treatment than before. There were BAL differential cell count abnormalities indicative of inflammation in both cases and controls.(ABSTRACT TRUNCATED AT 250 WORDS)     

Treatment of Moraxella bovis infections in calves using a long-acting oxytetracycline formulation

George, L.W. & Smith J.A. Treatment of Moraxella bovis infections in calves using a long-acting oxytetracycline formulation. J. vet. Pharmacol. Therap. 8, 55–61.
Studies were undertaken to determine the effectiveness of an oxytetracycline HCl formulation for the prophylaxis and treatment of chronic Moraxella bovis ocular infections in calves. Two separate experiments were performed. For the first, calves were separated into two groups and the eyes were infected with M. bovis. The eyes of these calves were observed and cultured for 37 consecutive days. On the 37th and 40th day, each of the five calves were treated intramuscularly with the drug (20 mg/kg of body weight). The other five calves (second group) remained untreated as controls. The cultures from the five treated calves were negative after the first antibiotic administration and remained so for 14 days. M. bovis was isolated from each eye of the control calves at least once during that time. None of the antibiotic-treated calves was completely resistant when reinfected with M. bovis. For the second experiment, calves were given a prophylactic administration of the formulation and were then infected with M. bovis 48 ( n = 4 calves) or 72 ( n = 4 calves) h later. These treatments resulted in a lower incidence of keratoconjunctivitis and a decreased duration of bacterial shedding, as compared to controls ( n = 8 calves), but did not completely prevent the occurrence of disease or the establishment of ocular infections.
Lisle W. George, Department of Medicine, School of Veterinary Medicine, University of California, Davis, CA 95616, U.S.A.     

Efficacy of tulathromycin in the treatment of bovine respiratory disease associated with induced Mycoplasma bovis infections in young dairy calves

The efficacy of tulathromycin in the treatment of bovine respiratory disease (BRD) due to Mycoplasma bovis was determined following experimental infection. Two highly pathogenic strains of M. bovis (with minimum inhibitory concentration values for tulathromycin of 1 and >64 microg/ml) were inoculated into 145 calves. Four days after inoculation, calves with clinical BRD were treated subcutaneously with saline or tulathromycin (2.5 mg/kg). Compared with saline, BRD-related withdrawals, peak rectal temperatures, and lung lesion scores were significantly lower for tulathromycin-treated calves (P < .01). Tulathromycin was highly effective in the treatment of BRD due to M. bovis in calves regardless of the minimum inhibitory concentration of the challenge strain (1 or >64 microg/ml).     

Experimental Mycoplasma bovis infection of the respiratory tract of calves

Multiple intranasal experimental Mycoplasma (M.) bovis infection was induced to 22 calves and resulted in clinical manifestations. M. bovis was re-isolated from 17 of 24 lungs which had been altered by pneumonia. Pasteurella haemolytica was the only pathogen isolated from the pathologically affected lung of one of the experimental animals. The haematogenic proliferation phase was successfully identified in 5 of 8 examined calves. Arthritis was not recordable from any of the experimental animals. Interstitial and catarrhal to catarrhal-purulent bronchopneumonias were the most common histological findings. Peribronchial lymphoid cell accumulations occurred frequently, as time passed from onset of infection. Antibodies against M. bovis were detected in blood serum of infected animals by means of film inhibition, as of the 3. week from infection, and by means of indirect haemagglutination, as of the 4. week from infection.     

The association between serological evidence of mycoplasma infection and respiratory disease in feedlot calves.

Calves from five Ontario feedlots were bled on arrival and approximately 28 days later. Calves treated during this interval for undifferentiated respiratory disease were classified as cases and untreated calves were classified as controls. Serum was titrated blindly for antibodies to Mycoplasma bovis and Mycoplasma dispar. Indirect hemagglutination titers of 1:20 or more were assumed to reflect recent or current exposure, whereas 1:10 or less were not. The titers to M. bovis increased in all feedlots indicating active infection. The initial titers to M. dispar were higher than the titers against M. bovis, yet they increased in all feedlots except one, suggesting widespread infection with this organism. There was an increased risk (although not statistically significant) of being treated if the titer against M. bovis rose during the period. Calves with low M. dispar titers on arrival were at increased risk of being treated and titer increases were strongly associated with treatment (statistically significant). Thus, the serological results indicate high prevalence of M. bovis and M. dispar in the feedlot calves and that calves with increasing titers in particular to M. dispar are at increased risk of being treated for respiratory disease.     

Pneumonia in calves: characterization of the bacterial spectrum and the resistance patterns to antimicrobial drugs

The population under study included young calves with pneumonia (group A, n = 13) and their controls (group B, n = 9), as well as older calves from which the lungs with (group C, n = 90) or without (group D, n = 10) lesions were collected after slaughter. Arcanobacterium pyogenes was the organism most commonly isolated from calves in group A (46%), followed by Haemophilus somnus (23%), Mannheimia haemolytica (15%), Streptococcus suis and Pasteurella multocida (7.7% each). Only S. suis (22%) and P. multocida (11%) were found in group B. P. multocida was isolated from 32% group C calves, H. somnus from 11%, A. pyogenes from 7.8%, M. haemolytica from 2.2% and S. suis from 1.1%. No specific pathogens were isolated in group D. Prevalence of Mycoplasma bovis infection was 69% in group A and 37% in group C. Ninety-eight strains were tested for resistAnce to antibiotics. Resistance to penicillin and ampicillin was present only in M. haemolytica (46%). High percentages of resistant strains were observed for streptomycin (48-100%), tetracycline (15-43%), sulfonamides alone (14-100%) or in combination with trimethoprim (0-100%). Therapeutic approaches to bacterial calf pneumonia in the area under study should be modified according to the isolated bacterial population, the observed antimicrobial resistances and the growing importance of Mycoplasma bovis.     

Mycoplasma agalactiae subsp. bovis in pneumonia and arthritis of the bovine.

The pneumonic lungs of 42 cattle from 26 feedlots were examined for the presence of mycoplasma, pathogenic bacteria and viruses. Four animals representative of two lots failed to yield mycoplasma. One of these yielded the virus of infectious bovine rhinotracheitis and Pasteurella hemolytica, the other yielded only P. P. multocida. Nine animals in eight lots yielded Mycoplasma sp.: five of these were M. bovirhinis, two were M. arginini and two were untypable. All of these animals yielded one or more of P. hemolytica, P. multiocida, infectious bovine rhinotracheitis virus or bovine virus diarrhea virus. Twenty-five of 29 animals in 16 lots yieled M. agalactiae subsp. bovis from lung tissues. The same organism was recovered from the arthritic joints of 12 of these animals. Eight of the 25 animals yielded no other pathogen and all of these had not received any treatment. Nine of the 25 M. agalactiae subsp. bovis positive animals also yielded one or more of P. hemolytica, P. multocida, Corynebacterium pyogenes or infectious bovine rhinotracheitis virus. Bacteriological and virological studies were not completed for the remaining eight of the 25 positive animals. In five lots of cattle which had not received medication for pneumonia and for arthritis only M. agalactiae subsp. bovis was recovered. Twenty-five grossly normal lungs obtained from normal cattle at the time of slaughter were cultured and all were negative. The possible role of M. agalactiae subsp. bovis in pneumonia and arthritis was discussed.     

Cytological findings in bronchoalveolar lavage fluid from feedlot calves: associations with pulmonary microbial flora.

Samples obtained by bronchoalveolar lavage (BAL) were used to evaluate pulmonary cytology in 59 feedlot calves with clinical signs of respiratory disease (cases) and 60 clinically normal comparison calves (controls). Many calves in both case and control groups had inflammatory changes in the lower respiratory tract, as determined by changes in proportions in the BAL differential cell count. Approximately 35% of cases and 40% of controls showed a normal differential cell count. It therefore appeared that the criteria used to select cases for treatment, which were similar to those often used in the field, were poor predictors of lower respiratory tract disease. A positive association was found between an increased proportion of neutrophils in BAL fluid and isolations of Pasteurella multocida and Mycoplasma bovis from BAL fluid.     

Investigation of factors of probable significance in the pathogenesis of pneumonic pasteurellosis in cattle.

Six groups of ten beef calves six to eight months of age were shipped from western Canada and observed untreated for one week after arrival. The following parameters were measured daily: body temperature, plasma fibrinogen, nasal bacterial mean colony counts of Pasteurella hemolytica and Pasteurella multocida, total and differential leukoyte counts, packed cell volumes and the following, twice during the week: serum and nasal antibody titres to P. hemolytica and parainfluenza-3 virus. The lungs from 44 of the calves were obtained at post mortem and given a numerical score based on the degree of pneumonia present. Animals were designated SICK and WELL according to body temperature and plasma fibrinogen. The SICK animals had higher nasal mean colony counts of P. hemolytica than the WELL animals. The SICK animals had lower levels of serum antibody to P. hemolytica than the WELL on day 1 but had a greater rise in titre over the week than did the WELL animals. Both groups were similar with regard to serum antibody to parainfluenza-3 virus and there was little change in these titres. The SICK animals had a much greater degree of pneumonia than the WELL. The values of some of the parameters were combined with the data of previously studied animals in order to provide a comparison of SICK and WELL with larger numbers of animals.     

Sub-clinical parasitism in spring-born, beef suckler calves: epidemiology and impact on growth performance during the first grazing season

Sub-clinical parasitism in spring-born single suckled beef calves was investigated from the middle of their first grazing season until weaning or housing later the same year. The study was conducted on four beef suckler herds in southern England over a 3-year period and involved a total of 334 spring-born beef suckler calves and their dams. The animals were grazed extensively on pastures naturally infected with nematode larvae. At the start of each period of observation, faecal samples were taken from calves and cows and subjected to routine worm egg counts; calves were re-sampled at the end of the grazing season.In July in each year and at each location the calves were ranked by initial weight within sex, paired according to rank and randomly allocated to either an untreated control group or a group in which the calves were each treated with an ivermectin sustained-release (SR) bolus. The calves in both trial groups, and their dams, were grazed together until weaning or housing. The calves were weighed at the initial allocation and at the end of the study. The adult cows were not treated with any anthelmintic during the study.The faecal nematode egg counts (FECs) conducted in July showed that the suckler cows were excreting worm eggs at low concentrations: range 0-100 eggs per gram (epg), with one individual count of 500epg, 88% of the cows sampled had counts of <50epg. Similarly, the counts from the calf samples were fairly low in July: range 0-250epg, 73% of the calves sampled had counts of <50epg. By the end of the grazing season, the faecal samples from the untreated control calves showed higher values: range 0-650epg, with only 58% having an epg of <50.The average rate of daily liveweight gain in the untreated heifer calves was 0.79kg per day, the corresponding figure for the heifer calves treated with the ivermectin SR bolus in mid-summer was 0.88kg per day; the difference of 90g per day was significantly different (P=0.0118). The average rate of daily liveweight gain in the untreated bull calves was 0.91kg per day, the corresponding figure for the bull calves treated with the ivermectin SR bolus in mid-summer was 1.01kg per day; the difference was significantly different (P=0.0169).     

Examination of the role of Mycoplasma bovis in bovine pneumonia and a mathematical model for its evaluation

The authors screened 34 large cattle herds for the presence of Mycoplasma bovis infection by examining slaughtered cattle for macroscopic lung lesions, by culturing M. bovis from lung lesions and at the same time by testing sera for the presence of antibodies against M. bovis. Among the 595 cattle examined, 33.9% had pneumonic lesions, mycoplasmas were isolated from 59.9% of pneumonic lung samples, and 10.9% of sera from those animals contained antibodies to M. bovis. In 25.2% of the cases M. bovis was isolated from lungs with no macroscopic lesions. The proportion of seropositive herds was 64.7%. The average seropositivity rate of individuals was 11.3% but in certain herds it exceeded 50%. A probability model was developed for examining the relationship among the occurrence of pneumonia, the isolation of M. bovis from the lungs and the presence of M. bovis specific antibodies in sera.     

Effect of bovine granulocyte colony-stimulating factor on the development of pneumonia caused by Mannheimia haemolytica

The role of recruited neutrophils in Mannheimia haemolytica infection is controversial. We hypothesized that the neutrophilia induced by recombinant bovine granulocyte colony-stimulating factor (GCSF) would lead to rapid bacterial clearance and less severe lesions after infection with M. haemolytica. Two experiments (A and B) were conducted in which four calves per experiment were treated daily with 5 microg/kg GCSF and four calves per experiment were treated with saline. All 16 calves were challenged with 5 x 10(9) colony-forming units (cfu)/ml (experiment A) or 4.5 x 10(8) cfu/ml (experiment B) of M. haemolytica bacteria, into the right bronchus by bronchoscope-placed catheter. The mean maximal blood neutrophil counts in non-GCSF-treated and GCSF-treated calves before bacterial challenge were 5.6 +/- 0.7 x 10(9)/liter and 25.4 +/- 2.7 x 10(9)/liter, respectively. Two untreated calves became neutropenic and were euthanatized 2 days after infection because of severe respiratory distress. GCSF-treated calves had a 37% reduction in lung lesions compared with nontreated calves, and this difference was significant (P=0.04) when the effect of previous antibody titre to leukotoxin was considered. The effect of GCSF treatment on the severity of clinical signs seemed to be influenced by the antibody titre to M. haemolytica leukotoxin, although this effect could not be conclusively addressed. In conclusion, GCSF induced neutrophilia and partially protected calves against experimental infection with M. haemolytica. These results imply that increased numbers of neutrophils may, under some circumstances, protect against severe pneumonia caused by M. haemolytica.