Prevalence of mycoplasmas in the respiratory tracts of pneumonic calves.

The prevalence of mycoplasmas in the respiratory tracts of 148 pneumonic calves originating from 25 herds in the Netherlands is reported. Four types of culture media were used to isolate mycoplasmas: solid modified Edward medium, 2 types of Friis media, and A7B differential agar medium. Mycoplasmas were isolated both from nasal swab specimens and lung lavage fluids collected from live calves and from nasal mucosa and lung tissue specimens collected post mortem. All of the mycoplasma strains isolated could be identified as either Ureaplasma diversum (isolated from 80% of 25 herds), Mycoplasma dispar (92%), M. bovirhinis (88%), M. bovis (20%), M. bovigenitalium (4%), M. arginini (16%), or M. canis (12%). Isolation rates of M. dispar and U. diversum were considerably higher from lung lavage fluids than from nasal swab specimens. M. bovis was detected only in fattening herds and not in dairy herds. The respiratory tracts of 75% of the calves examined contained at least 2 mycoplasma species. In total, 25 different combinations of mycoplasma species were detected in specimens collected from noses and lungs. The pathogenic species U. diversum and M. dispar had not been isolated before in the Netherlands.     

Transmission of Mycoplasma dispar among calves collected and reared for beef production

Fortytwo calves, 28 to 117 days old, were collected from 23 dairy farms and transported in a lorry, allowing direct contact between the calves, to 8 calf rearing farms. The average transport time per calf was 4.5 h, ranging from 0.3 to 12.8 h. The calves were sampled by nasal swabbing for mycoplasmas first before loading and then immediately after transport. Thirteen of the calves were transferred to farm I. They were placed in individual pens in a separate room to themselves, and were sampled at intervals for a period of 4 weeks.Ten of the 42 calves (23.8 %) originating from 5 of the source farms were found initially positive for M. dispar with titers > 4 log₁₀ ecu; 3 of these 10 calves were delivered to farm I and 7 calves to 6 others of the 8 receiving farms. Three initially infected calved delivered to farms I continued to be positive throughout the follow-up period; among the 10 initially negative calves the frequency of detected infection, and the geometric mean titer (within parenthesis), developed so that on days 1, 7, 14, and 28 the figures were: 2 (2.5), 8 (4.3), 9 (4.7), and 10 (5.5), respectively.After transport 3 initially negative calves were found positive with low titers. Two of them were placed on farm I. In one of them positivity proved to be only transient; the case seems to represent a phenomenon of transfer of mycoplasmas without establishment of infection. In contrast, at least 4 (possibly 7) calves, negative both before and after transport–ascribed above to the group of 10 initially negative calves arriving on farm I–had in all likelihood caught the infection during the transport. Two of the 10 calves most likely caught the infection on the farm; for 3 calves the evidence was equivocal as to the 2 alternatives.Seven of the 42 calves (16.7 %) were found to be initially infected with M. bovir-hinis, 2 of the 42 with Acholeplasma laidlawii. Among the 13 calves transported to an reared on farm I, 8 were found to be positive at least once for M. bovirhinis during the study. Colonisation by this mycoplasma was partly detected only intermittently and the detectable prevalence among the 13 calves at its highest was only 38.5 %.     

Colonisation pattern of the respiratory tract of calves by Mycoplasma dispar

Eight conventionally reared, 1- to 11-week old Ayrshire calves were naturally infected by a strain of Mycoplasma dispar (M. dispar). The colonisation was quantitatively followed by nasal swab samples, transtracheal aspiration samples and by the examination of the whole of the respiratory tract for mycoplasmas at slaughter after a follow-up period of 7–10 months.The fairly uniform pattern of the colonisation by M. dispar was revealed: A high degree of colonisation, measuring 10⁵–10⁸ colour change units (ccu) per nasal sample, lasted for a period of 2–5 months and was followed by a slow decrease in titres. Seven of the calves still harboured M. dispar in their respiratory tracts at slaughter. Intermittently obtained transtracheal aspiration samples were all positive for M. dispar and the titres were regularly higher than those for the simultaneously taken nasal samples indicating a high ability of M. dispar to continuously colonize the more distal parts of the respiratory tract. It was demonstrated that the sensitivity of nasal swabbing in the detection of M. dispar infection largely depended on the phase of colonisation : The method was good for the detection of a fairly recent infection of M. dispar, but inadequate for detection of low grade carriers.In various phases, the calves also became infected by Mycoplasma bovirhinis and Acholeplasma laidlawii. Their ability to colonize the whole respiratory tract was lower than that of M. dispar.     

The nasal mycoplasmal flora of healthy calves and cows.

The nasal mycoplasmal flora of 270 healthy cows from 27 herds in the Netherlands and 35 healthy calves from 7 of these herds was examined. Various methods for isolating mycoplasmas were compared. The prevalence of the various species was as follows: Ureaplasma diversum in 3 (9%) calves; Mycoplasma dispar in 14 (40%) calves; M. bovis in 1 (3%) calf; M. bovirhinis in 23 (66%) calves and 16 (6%) cows; M. bovoculi in 8 (23%) calves and 53 (20%) cows; M. canis in 1 (3%) calf; M. equirhinis in 2 (1%) cows; M. conjunctivae in 2 (1%) cows; Acholeplasma laidlawii in 1 (3%) calf and 3 (1%) cows; and A. axanthum in 7 (3%) cows. The noses of healthy calves were less frequently colonized by the pathogenic species U. diversum and contained fewer U. diversum and M. dispar organisms than the noses of pneumonic calves. We concluded that the mycoplasmal flora of calves and healthy cows was quite different and also that cows play only a minor role in the epidemiology of pathogenic mycoplasma species of calves in the Netherlands.     

The incidence of Mycoplasma dispar, Ureaplasma and conventional Mycoplasma in the pneumonic calf lung.

This report describes the incidence of Mycoplasma dispar, ureaplasma and conventional (large colony) mycoplasma isolated from the pneumonic lungs of groups of young calves and the identification to species level of mycoplasmas in mixed populations with the aid of the indirect fluorescent antibody test. Pneumonic lung tissue yielded one or more mycoplasma species from 88% of the 153 calves cultured. The mycoplasmas identified and percent of the calves with lungs positive for each species were: M. dispar (56%), ureaplasma (44%), Mycoplasma bovis (37%), Mycoplasma arginini (33%) and Mycoplasma bovirhinis (23%). Conventional mycoplasmas isolated from two calves (1%) could not be identified using the antisera available.     

Effects of age, environmental temperature and relative humidity on the colonization of the nose and trachea of calves by Mycoplasma spp

Nasal and tracheal swabs sequentially collected from three groups of eight calves between the ages of 1 and 98 days indicated that the nose and trachea were colonized by Mycoplasma spp. during the first weeks of life. Over 92% of all calves harboured Mycoplasma spp. in their noses when they were 2 weeks old, the rate of recovery falling gradually thereafter. The peak period of recovering mycoplasmas from the noses and tracheas of calves was at 6 weeks old. M. bovirhinis, M. arginini and Acholeplasma laidlawii predominated in the nose while M. dispar and M. bovirhinis predominated in the trachea. There was no association between rates of isolation and clinical signs of respiratory disease. There were no significant differences between the frequencies of isolation of Mycoplasma spp. from groups of calves kept under different environmental temperatures and relative humidities.     

Use of a polymerase chain reaction for detection of Mycoplasma dispar in the nasal mucus of calves.

A polymerase chain reaction (PCR) assay was developed for the detection of Mycoplasma dispar in nasal mucus samples collected from calves. The target DNA sequence was the 16S rRNA gene, and the fragment was selected within a region of high polymorphism. The specificity and detection limit of the method were determined. This method was then used for the detection of M. dispar in nasal swabs collected from 301 calves, including 155 clinical samples from animals showing signs of respiratory disease and 146 samples from healthy animals. PCR with generic primers was applied to the detection of Mollicutes, followed by the detection of M. dispar. Mollicutes were detected in 52.05% of clinical samples from healthy animals and in 90.96% of samples from sick animals. Mycoplasma dispar was detected in 6.16% of healthy animals and in 34.84% of sick animals. The PCR assay was useful in verifying the presence of M. dispar in calves and may be a useful tool in monitoring this mycoplasma in cattle herds.     

Isolation of mycoplasma species from the lower respiratory tract of healthy cattle and cattle with respiratory disease in Belgium

Between 1997 and 2000, a total of 150 healthy cattle and 238 animals with respiratory disease were examined for six Mycoplasma species. Attempts were made to detect Mycoplasma canis, Mycoplasma dispar and Ureaplasma diversum in calves with recurrent disease, and all three of these species were identified in calves with recurrent disease and in healthy lungs. In healthy calves, 84 per cent of bronchoalveolar lavage fluids were mycoplasma free; when cultures were positive, Mycoplasma bovirhinis was the only species isolated. Mycoplasmas were isolated from 78 per cent of animals suffering recurrent respiratory disease and from 65 per cent of acute respiratory cases. Mycoplasma bovis was isolated from bronchoalveolar lavages from 35 per cent of calves suffering recurrent respiratory disease, and from 50 per cent of acute cases, and from 20 per cent of pneumonic cases examined postmortem. M bovis was associated with other Mycoplasma species in 44 per cent of cases. M dispar was also isolated from 45.5 per cent of calves suffering recurrent respiratory disease, often in association with M bovis. M canis was identified for the first time in diseased Belgian cattle. Other mycoplasmas, including Mycoplasma arginini, Mycoplasma alkalescens and U diversum, were isolated less frequently. Associations between mycoplasmas and other pathogens were often observed. Among lungs infected with Pasteurella and/or Mannheimia species, more than 50 per cent were mixed infections with M bovis.     

Pathogenicity of some Mycoplasma and Acholeplasma species in the lungs of gnotobiotic calves

Cloned cultures of 16 strains, representing nine different species of Mycoplasma and Acholeplasma, were inoculated intratracheally into gnotobiotic calves. Strains of M bovirhinis, M canadense, M verecundum, A axanthum and A modicum did not produce visible pneumonic lesions and were not reisolated from the lungs. Strains of M alkalescens and M arginini colonised the lower respiratory tract but failed to produce visible pneumonia. M bovigenitalium (strain M991/70) and M dispar (strain Gri226) both colonised the respiratory tract and induced pneumonic lesions estimated to involve up to 8 per cent (M bovigenitalium) and 17 per cent (M dispar) of the lung. Histologically M bovigenitalium produced a cuffing pneumonia and M dispar produced a interstitial alveolitis.     

Transmissibility of Mycoplasma dispar under experimental conditions

Twenty three 1- to 18-week old Ayrshire calves were exposed to infection by Mycoplasma dispar, 1 or 2 at a time, in a series of consecutive experiments. Exposure took place either at direct contact, i. e. with the possibility of physical contact between susceptible and infected calves (type I), or through separation of susceptibles from the nearest infected calf at a distance of 0.8 m (type II) or 1.3 m (type III). Frequent nasal swabbing was used for the detection of the infection and the measurement of the level of colonisation. Ten, 4, and 9 calves were subjected to type I, II, and III exposures, respectively. When the first positive nasal specimen preceding a rise in titres in the following specimens was used to signify an establishment of infection of a susceptible calf, the medians (ranges) of the times for the transmission of infection (including the latent period of early colonisation undetectable by nasal swabbing) in the 3 exposure groups were: 4.5 (1-27), 8.5 (7-9), and 17 (6-32), respectively. The difference between groups I and III was significant (p < 0.05). The last figures for type III of exposures represent the results of only 5 calves succesfully exposed; the remaining 4 of the 9 calves of this exposure type were not found to pick up the infection within the periods of 2 to 3 weeks of exposure. The results indicate a transmission mode via droplets.Four calves exposed only for 4 h by direct contact were found to be infected on days 0, 4, 4, and 7, respectively. This indicated variability in the length of the latency as regards the detection by nasal swabbing.Four of the 7 calves in the type I experiments sampled at hour 4 showed low titers for M. dispar. Only in 1 of these calves was this early finding of transfer followed by positivity and a rise in titers through subsequent specimens. The other 3 calves became positive anew, with subsequent developing colonisation, first on days 4, 4, and 12, respectively.     

Increasing prevalence of Mycoplasma bovis in Danish cattle

A study on the prevalence of mycoplasmas in pneumonic bovine lungs was performed on material submitted for diagnostic purposes at the Danish Veterinary Laboratory, Copenhagen. Among the 50 examined cases 43 (86.0%) were found to be infected with mycoplasmas. The predominant mycoplasmas were Ureaplasma spp. (72.0%), M. dispar (48.0%) and M. bovis (24.0%). Other mycoplasmas were M. bovirhinis (20.0%) and M. bovigenitalium (6.0%). Among the infected lungs multiple species infections were predominant (76.7%) over single species infections (23.3%) with M. dispar-Ureaplasma (25.6%), M. bovis-Ureaplasma (18.6%) and M. dispar-M. bovirhinis-Ureaplasma (11.6%) infections being the most frequently encountered combinations. There appears to be an increasing prevalence of M. bovis (24.0%) as compared to earlier reports (0.6-2.0%), thus calling for special attention upon this mycoplasma. Pulsed field gel electrophoresis (PFGE) analysis of 11 field isolates of M. bovis from 9 different farms revealed different profiles except for 2 isolates which were recovered from the same farm. Because mycoplasmas belonging to the 'M. mycoides cluster' were not encountered during this study; it appears that the Danish cattle population is still free from this group of mycoplasma in spite of their presence in some other European countries.     

Identification of Mycoplasmatales in pneumonic calf lungs

Lungs from 153 calves with clinical signs of pneumonia were examined post-mortem (PM) for the presence of mycoplasmas and ureaplasmas during a 38-month period. Sixty-two percent of the cases were submitted during the months when wide fluctuations in climatic conditions occur. Using indirect fluorescent antibody tests (IFAT) and culture, mycoplasmas and/or ureaplasmas were detected in 63% of the lungs examined. Mycoplasma dispar was detected in 39%, M. bovis in 36%, Ureaplasma spp. in 22% and M. bovirhinis in 8.5% of the lungs. Thirty percent of the lungs were infected with more than one species; the most frequent combination was M. bovis, M. dispar and Ureaplasma spp. (10.5%). M. arginini, M. bovigenitalium and acholeplasmas were not cultured. M. dispar was shown to remain viable for up to 15 days PM in apical and cardiac lobes held at 4 degrees C and also was detected by IFAT in the same tissues for 49 days.     

Isolation of Mycoplasmas from nasal swabs of calves affected with respiratory diseases and antimicrobial susceptibility of their isolates

Nasal swabs of 293 calves were examined for Mycoplasma. The samples were collected from calves affected with respiratory diseases on 71 farms in various parts of Japan between 1996 and 1997. Mycoplasma bovirhinis was isolated from 47 of 293 calves (16.0%). Mycoplasma alkalescens, M. bovis, M. arginini, M. bovigenitalium and Acholeplasma spp. were isolated from 19 (6.5%), seven (2.4%), four (1.4%), four (1.4%) and 18 (6.1%) calves, respectively. Pasteurella multocida and P. haemolytica were isolated from 60% of Mycoplasma-positive calves. However, other bacteria were not isolated from calves. To evaluate the antimicrobial susceptibility of their isolates, 68 M. bovirhinis, 21 M. alkalescens and 10 M. bovis strains were examined for 12 antimicrobial agents. All isolates showed higher susceptibility to tiamulin than to the other drugs used in the study. However, erythromycin had no effect on any of the Mycoplasma strains studied. The field isolates were less susceptible than the type strains to some drugs, such as spiramycin, oxytetracycline and tylosin.     

Prevalence of Giardia spp. infection in pre-weaned and weaned calves in relation to management factors

Two hundred and forty calf faecal samples from 16 Malaysian farms were screened by PCR for Giardia spp. The overall prevalence was 12.5% and the overall farm prevalence was 68.8% (11/16 farms). The prevalence in pre-weaned and weaned calves was 16.7% and 8.3%, respectively. Sequence analysis of 25 isolates identified all as G. duodenalis assemblage E. Management factors associated with an increased risk of infection with Giardia spp. included keeping weaned calves in pens with sand floors and calf age. Keeping pre-weaned calves in pens with concrete floors and calving in single cow calving areas decreased the risk.     

Epidemiologic Survey and Species Determination of Coccidia Infection in Dairy Cattle in Yuzhong,Gansu Province

This investigation aimed to examine coccidia infection in diary cattle in Yuzhong,Gansu province.An investigation was carried out on the Holstein calves less than one year old in six large-scale dairy farms.A total of 234 fecal samples were examined,and oocysts were identified to the species level on the basis of morphological features after positive samples were mixed thoroughly with 2.5% potassium dichromate solution.The results showed that the prevalence rate of coccidia in Holstein calves reached 46.15%.The average prevalence rate of the calves under 3 months old was 52.43%,the 4 to 6 months old calves was 37.50% and the 7 to 12 months old calves was 43.37%.Seven species of Eimeria coccidians and one species Isospora sp.including Eimeria bovis,Eimeria auburnensis,Eimeria zuernii,Eimeria alabamensis,Eimeria subspherica,Eimeria ellipsoidalli,Eimeria canadensisi,were identified in the investigation,and Eimeria bovis,Eimeria auburnensis,Eimeria zuernii were the dominant species.In conclusion,the prevalence rate of coccidiosis in dairy cattles in Yuzhong,Gansu province was high.Therefore,appropriate strategies and measures should be taken to control coccidiosis prevalence in dairy cows in this region.     

甘肃榆中县奶牛球虫病流行病学调查与虫种鉴定

奶牛球虫病呈世界性分布,中国部分省市也有报道,但是甘肃省至今尚未有关于奶牛球虫病的相关调查。甘肃省榆中县是甘肃省奶牛主产区之一,为了解甘肃省榆中县奶牛球虫感染情况,对当地6个规模化奶牛养殖场1岁以内荷斯坦奶牛犊牛球虫感染情况进行了调查。试验采用显微镜下观察的方法检测了该地区的234份奶牛粪便样品,并对球虫阳性粪便进行重铬酸钾法孵化来鉴定球虫种类。结果发现,犊牛球虫总感染率达到46.15%,其中1~3月龄犊牛平均感染率为52.43%,4~6月龄犊牛平均感染率为37.50%,7~12月龄犊牛平均感染率为43.37%。经虫种鉴定,共发现7种艾美尔属球虫和1种等孢属球虫,7种艾美尔属球虫分别是牛艾美尔球虫(Eimeria bovis)、奥博艾美尔球虫(Eimeria auburnensis)、邱氏艾美尔球虫(Eimeria zuernii)、阿拉巴艾美尔球虫(Eimeria alabamensis)、亚球形艾美尔球虫(Eimeria subspherica)、椭圆艾美尔球虫(Eimeria ellipsoidalli)、加拿大艾美尔球虫(Eimeria canadensis),其中牛艾美尔球虫、奥博艾美尔球虫、邱氏艾美尔球虫为优势种。本次调查结果表明,甘肃省榆中县奶牛球虫感染情况严重,应引起相关部门重视。     

Mycoplasma species and related organisms isolated from ruminants in Britain between 1990 and 2000

Between 1990 and 2000, more than 1600 mycoplasmas and the related acholeplasmas were identified from ruminant animals by the Mycoplasma Group at the Veterinary Laboratories Agency--Weybridge. Mycoplasma bovis was the most commonly identified pathogen, mostly from pneumonic calves but occasionally from cattle with mastitis and arthritis. Mycoplasma canis was first isolated in Britain in 1995 from pneumonic calves and the number of isolates increased to 18 per cent of the total mycoplasmas isolated from cattle in 1999. The ELISA for antibodies to M. bovis detected 1971 positive samples (22 per cent) among 8959 serum samples, mainly from pneumonic cattle. Other mycoplasmas identified included Mycoplasma dispar from the lungs of cattle with respiratory disease, and Mycoplasma bovigenitalium from the reproductive tract of cows with vulvovaginitis and infertility. Mycoplasma bovirhinis and Acholeplasma species were found commonly but are thought to be more opportunistic than pathogenic. In sheep and goats, the majority of Mycoplasma species isolated were identified as Mycoplasma ovipneumoniae from pneumonic sheep, Mycoplasma conjunctivae from sheep with keratoconjunctivitis, and the ubiquitous Mycoplasma arginini.     

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.     

Studies of bovine mycoplasma mastitis. 1. Review of literature on the occurrence of bovine mastitis with Mycoplasma involvement]

The clinical pattern as well as the pathologico-anatomic or histological changes due to mycoplasma mastitis are neither specific nor pathognomic. Mastitis pathogens so far described included M. bovis, M. bovigenitalium, A. laidlawii, A. axanthum, M. alkalescens, M. canadense, M. dispar, M. bovirhinis, strains of Group 7 according to Leach, strain ST 6, and ureaplasma strains. In the GDR, enzootic mastitis has been confined to A. laidlawii and A. axanthum.     

Prevalence of antimicrobial resistance among Salmonella on midwest and northeast USA dairy farms

The objective of this study was to determine the prevalence of antimicrobial resistance among Salmonella isolated from dairy herds in New York, Minnesota, Michigan, and Wisconsin, USA. Serogroup and antimicrobial susceptibility characteristics were determined for Salmonella from cattle and environmental samples collected during August 2000–October 2001 as part of a longitudinal study where 129 herds were visited at 2-month intervals. Salmonella isolates were tested (using a broth microdilution method) for susceptibility to amoxicillin/clavulanic acid, ampicillin, ceftiofur, ceftriaxone, cephalothin, chloramphenicol, ciprofloxacin, gentamicin, kanamycin, nalidixic acid, streptomycin, sulfamethoxazole, tetracycline, and trimethoprim/sulfamethoxazole. Of the 1506 isolates tested for minimum inhibitory concentrations to these 14 antimicrobial agents, 81.2% were pan-susceptible and for most herds (81.6%) the predominant antimicrobial resistance pattern was pan-susceptible. At least 1 Salmonella isolate resistant to 5 or more antimicrobial agents was found on 23.6% of herds. This resistance phenotype was most common among serogroups B and E1 and among samples from calves and farmer-designated sick cows. Resistant samples most frequently exhibited resistance to tetracycline, streptomycin, and/or ampicillin. No samples were resistant to ceftriaxone (though 13 were in the intermediate range), and very few samples were resistant to ciprofloxacin (n = 1), nalidixic acid (n = 5), or trimethoprim/sulfamethoxazole (n = 7).