Detection of chlamydiae in boar semen and genital tracts

Chlamydiae cause abortion and reproductive disorders in sows. Although organisms can infect the male genital tract, little is known about the disease situation in boars. Hence, we examined the prevalence of chlamydial infection in semen and genital tracts of boars. Samples collected from Swiss boars (group A: n=42), and boars from Germany (group B: n=39) were examined by bacteriology, LPS-ELISA, immunohistochemistry (IHC) and polymerase chain reaction (PCR). The latter methodology involved use of three PCR assays including 16Sig rDNA, IGS-S (intergenic spacer 16S/23S-Short) and IGS-L (intergenic spacer 16S/23S-Long) PCR for comparison methods. PCR sensitivity and the presence of potential PCR inhibitors were determined by spiking semen with Chlamydophila (Cp.) abortus DNA. Detection limits of the 16Sig and IGS-S PCR were 10 templates, while the IGS-L PCR was less sensitive (100 templates). Of 25 semen samples that were collected from group A, one semen sample was positive for Cp. psittaci and two were positive for Chlamydia-like organisms by 16Sig PCR. Screening of sera from Swiss boars revealed three animals with positive reactions in the LPS-ELISA, although we failed to detect chlamydiae within organs of these or sera-negative animals by IHC or IGS-S PCR. In group B, 10 ejaculates were positive for Chlamydia (C.) suis and two were positive for Chlamydia-like organisms by 16S PCR. The identification of DNA from Chlamydia-like organisms in semen from both groups of boars was surprising and a role for these bacteria in reproductive diseases requires further assessment. In conclusion, the prevalence of chlamydial infection was low in group A animals indicating that venereal transmission may not be significant for Chlamydia-associated reproductive diseases in pigs, although rare cases may occur.     

Actinobacillus pleuropneumoniae infections in closed swine herds: infection patterns and serological profiles

Many farrow-to-finish herds are endemically infected with Actinobacillus pleuropneumoniae. In order to control the disease efficiently, a better knowledge of the ages at which pigs become infected is necessary. Furthermore, no information is available concerning the influence of maternally derived antibodies on the colonization of the upper respiratory tract. Therefore, A. pleuropneumoniae infection patterns were studied in five farrow-to-finish pig herds (A-E) with a history of pleuropneumonia. A longitudinal study was carried out in herds A and B. In these herds, piglets from sows carrying A. pleuropneumoniae in their noses or tonsils were sampled. Nasal and tonsillar swabs as well as sera, were collected from these animals at the age of 4, 8, 12, 16 (herds A and B) and 23 weeks (herd B). At these ages other pigs from the same sows were euthanized. The lungs were macroscopically examined and samples from nose, tonsils and lungs were collected at necropsy. A cross-sectional study was performed in herds C-E. In these herds nasal and tonsillar swabs, as well as sera, were taken from 10 animals of 4, 8, 12 and 16 weeks of age. Lung, nasal and tonsillar samples were tested for the presence of A. pleuropneumoniae by routine bacteriology and PCR with mixed bacterial cultures. The sera were examined for the presence of Apx toxin neutralizing antibodies. In herd A, A. pleuropneumoniae serotype 2 and 10 strains were isolated, whereas serotype 2, 3, 5b and 8 strains were demonstrated in herd B. In most herds, A. pleuropneumoniae was detected in mixed bacterial cultures of tonsillar and/or nasal samples by PCR from the age of 4 weeks onwards. Colonization of the lungs and development of lung lesions was observed in 12- and 16-week-old animals of herd A and 23-week-old animals of herd B. In most herds, high antibody titres were detected in 4-week-old piglets. These titres decreased during the first 12 weeks of age, but thereafter, increased. It was concluded that PCR with mixed bacterial cultures from tonsillar swabs is a valuable tool for the detection of infected animals. It was also concluded that colonization of tonsils and nasal mucosae can occur in the presence of maternally derived antibodies. Infection of the upper respiratory tract without lung involvement did not result in development of Apx toxin neutralizing antibodies. Therefore, such serological assays cannot be used for the detection of subclinically infected animals.     

Prevalence of Chlamydiaceae and Mollicutes on the genital mucosa and serological findings in dairy cattle

It was the aim of this project to obtain information on the prevalence of Chlamydiaceae and Mollicutes and their potential importance for reproductive problems in cattle. Cervical or vaginal swabs were taken from 644 animals in 196 farms and blood samples were collected from 375 cattle. Out of the animals, 6.8% had aborted within the last 12 months, 2.6% showed clinical vaginitis and 11.6% clinical endometritis. Chlamydiaceae were detected and identified by PCR followed by restriction fragment length polymorphism (RFLP) analysis. For the detection and identification of Mollicutes cultivation procedures, biochemical differentiation and serological identification were used. Sera were tested for antibodies against Chlamydiaceae and Mycoplasma (M.) bovis by ELISA and against M. bovigenitalium by Western blot analysis. Chlamydophila (Cp.) abortus was found in three cervical swabs. Cp. pecorum was detected in 9% of cervical or vaginal swabs. The majority of Cp. species found was Cp. pecorum and thus fertility problems caused by Cp. abortus are limited. M. bovis was found in only one genital swab. M. bovigenitalium was rarely diagnosed (3% of cervical and 2% of vaginal swabs). M. bovigenitalium was found more often in cattle having aborted (4/32 animals) than in cattle without history of abortion (5/220, p<0.05). Ureaplasma (U.) diversum existed in 12% of cervical and 36% of vaginal swabs and was found in 8 out of 17 animals with vaginitis. Out of the animals tested, 44.9% were seropositive for Chlamydiaceae, 14.8% for M. bovis and 27.3% for M. bovigenitalium.     

Comparison of methods for the sampling and isolation of toxigenic Pasteurella multocida from the nasal cavity of pigs

The efficacy of detecting toxigenic Pasteurella multocida from nasal swabs of slaughtered and live pigs was assessed. The isolation of toxigenic P multocida from nasal cavities of slaughtered bacon pigs from two herds with atrophic rhinitis was reduced by immersion in the hot water tank by 25 per cent and 75 per cent. Individual sows from one of the infected herds were repeatedly swabbed to find the best method of isolating toxigenic P multocida. Toxigenic P multocida were isolated from 50 per cent of cotton swabs inoculated on to selective medium the same day. After 24 hours in the post, 45 per cent of cotton swabs placed in transport medium, 38 per cent of alginate swabs dissolved in transport medium and inoculated into mice, and 36 per cent of the dissolved swabs inoculated directly on to selective medium yielded toxigenic P multocida. These bacteria were isolated from only 25 per cent of cotton swabs held in transport medium at 10 degrees C for 48 hours to simulate prolonged postage times; from slaughtered pigs a similar reduction in isolation was seen with swabs kept for 24 or 48 hours. The reduced isolation caused by a delay before culture was associated with an overgrowth of other flora. The development of this flora was prevented by storage of swabs at 4 degrees C in the laboratory or by the use of cool boxes for postage.     

Prevalence of respiratory pathogens in diseased, non-vaccinated, routinely medicated veal calves

The prevalence of respiratory pathogens in diseased veal calves was determined in 24 respiratory disease outbreaks in 15 herds in Belgium. Bacteria were cultured from nasopharyngeal swabs and seroconversion against viruses and Mycoplasma bovis was determined on paired sera. At the individual calf level, Mycoplasma species, Mannheimia haemolytica and Pasteurella multocida, were isolated from 70.5 per cent, 21.5 per cent and 26.0 per cent of swabs, respectively. At the herd level, the presence of M bovis could be confirmed in 84.6 per cent of the herds examined. Seroconversion against bovine viral diarrhoea virus (BVDV) was present in 71.4 per cent of herds, parainfluenzavirus type 3 in 53.3 per cent, bovine respiratory syncytial virus in 40.0 per cent, bovine adenovirus type 3 in 46.7 per cent, bovine coronavirus in 30.0 per cent, and bovine herpesvirus type 1 in 26.7 per cent. At postmortem examination, Mycoplasma species could be cultured from 61.9 per cent of pneumonic lungs (n=21). Sixty per cent of calves tested were positive for BVDV (n=20), and 20.0 per cent were positive for bovine respiratory syncytial virus (n=16).     

Prevalence of Streptococcus suis types 1 and 2 in domestic pigs in Australia and New Zealand

Using an indirect fluorescent antibody test, 54 per cent of 734 palatine tonsils of conventional pigs slaughtered in Australia and New Zealand were found to be infected with Streptococcus suis type 1 and 73 per cent of 959 were infected with S suis type 2. Variations in the prevalence of infection in pigs from different herds were thought to be due to differences in the sample sizes rather than to real differences in the prevalence between herds. The prevalence of infection with S suis was similar in pigs of either sex and in different age groups. Streptococcus suis type 2 was detected in the blood of 3 per cent of apparently normal pigs slaughtered at a meat processing plant. The presence of this organism in edible tissue may pose a health risk to consumers and meat-workers. Both S suis types 1 and 2 were detected in the vaginas and uteri of slaughtered pigs and the female reproductive tract could be another site for the carriage of infection. Piglets from sows with vaginas infected with S suis type 2 became infected earlier than piglets from sows with uninfected vaginas. No infected male reproductive tracts were detected and venereal transmission of S suis therefore appears unlikely. Three specific pathogen free herds were found to be free from infection with both S suis types 1 and 2. It is concluded that hysterectomy derived piglets are delivered free from infection, whereas some piglets born to sows with uterine and vaginal infections are either born infected or become infected at, or soon after, birth.     

Causes of culling and death in sows.

Over a 4-year period, the annual number of culled sows in 9 Danish herds averaged 54.8 per cent of the year-sows and the number of culled sows in per cent of total number of farrowings averaged 25.8 (Table I). -- The culling rate varied considerably from herd to herd within the same year and from year to year (Tables I and II); however, the average annual culling rate for all the herds only presented small variations (Table II). The average number of litters reared per sow before culling was 3.6. The culling rate was higher in pedigree herds than in commercial herds, and it was highest in the small pedigree herds (Table III). The hygiene level in the herds and the introduction of new female breeding stock did not influence the culling rate (Table IV). A proportionally lower percentage of the sows was culled in herds where the dry and pregnant sows were housed in stalls and/or were tethered, as compared to herds where these sows were housed in pens (Table V). -- The culling rate in the age groups of sows with less than 8 farrowings remained at approximately the same level (Table VI). The main reason for culling was infertility problems, which were recorded in 41.4 per cent of the culled sows, while 16.7 per cent of the sows were culled because their litters were poor and/or small (Table VII). The mortality rate among the culled sows was 11.9 (Table VII), and the main causes of death were chronic pyogenic infections, which occurred in 25.5 per cent of the fatal cases (Table VIII). Certain aspects concerning the recording and calculation of culling rates in the different herds are discussed and it is emphasized that the culling rate per se may not have any direct relationship to the productivity in the herd.     

Salmonella in sows: a longitudinal study in farrow-to-finish pig herds

Besides finishing pigs, sows are also believed to be important in the epidemiology of Salmonella. The study objective was to investigate the prevalence of Salmonella excretion in sows during an entire reproductive cycle. In 3 farrow-to-finish herds, groups of 34, 40 and 32 sows, respectively, were sampled serially. Faecal samples, environmental swabs and feed samples were taken and submitted to a qualitative Salmonella isolation. All isolates were characterised using RAPD and a representative number of isolates was serotyped. The prevalence of Salmonella excretion was < 10% during gestation, around farrowing and during lactation, but a significant increase in the number of Salmonella excreting sows was found in herds A (p < 0.01) and C (p = 0.02) after weaning. S. Infantis was the most prevalent serotype in herd A, S. Derby in herds B and C. Except for the S. Infantis group in herd A, all isolates within each group of the RAPD analysis belonged to the same serotype. Three sows in herd A and 1 sow in herd C shed different serotypes at different time points. The present results indicate that sows can maintain Salmonella infections in farrow-to-finish herds and that culled sows, leaving the herd after weaning, may constitute a substantial risk for contamination of their carcasses with Salmonella.     

Study of agglutinins to Brucella abortus, B canis and Actinobacillus equuli in horses

Horses at a veterinary teaching hospital and a slaughterhouse were surveyed for antibodies to Brucella abortus, B canis and Actinobacillus equuli. Four of the 141 hospitalised horses and none of the 73 slaughtered horses had titres of 1:100 or greater to B abortus. Six horses of both populations reacted to the card test. One was culture positive. A card test using B canis antigen was positive in 38 per cent of the sera from hospitalised horses and all of the slaughtered horses. Twenty (27.4 per cent) of the latter group had high tires in a tube agglutination test. High titres could not be reduced by 2-mercaptoethanol serum treatment. The titres appeared to be associated with advanced age but not to sex. Adsorption of sera with B canis did not affect titres to A equuli but the reverse was true.     

Prevalence of Coxiella burnetii infection in Dutch dairy herds based on testing bulk tank milk and individual samples by PCR and ELISA

A study using an ELISA and a real-time PCR assay based on the detection of the repetitive transposon-like gene of Coxiella burnetii revealed that infection with the bacterium was widespread among Dutch dairy herds, with antibodies detected in bulk tank milk (BTM) from 268 of 341 herds (78.6 per cent) and bacterial DNA detected in 193 of 341 herds (56.6 per cent). The BTM samples were taken in November and December 2007. Serological and molecular studies in young and adult cattle selected from 100 herds showed that antibodies were present in the blood of 470 of 2936 (16.0 per cent) lactating cows but only in 19 of 1831 (1.0 per cent) young animals. Bacterial DNA was detected in the milk of 254 of 2925 (8.7 per cent) lactating cows; bacterial DNA was not detected in any of the faecal samples obtained from youngstock. The blood and milk samples were taken from the cattle in the period January to April 2008.     

Isolation of leptospires from the genital tract and kidneys of aborted sows

Twenty sows were examined as part of an investigation into the relative importance of the genital and urinary tracts as sites of leptospiral persistence in pigs. The sows had previously either aborted leptospire-infected fetuses or aborted on farms where leptospire infection had been demonstrated in other litters. Leptospires belonging to the Australis serogroup were recovered from the upper genital tract of all the sows. In contrast, leptospires were only recovered from 75 per cent of their kidneys. Isolates from 10 of the sows were identified to serovar level: six were serovar muenchen and four were bratislava. Persistence was observed in renal and genital tissues for up to 147 days after abortion.     

集约化养猪场嗜流产衣原体病流行状况的初步调查

本试验对北京郊区5个规模化猪场断奶仔猪、育成猪、育肥猪和怀孕母猪采集血清共507份,同时采集密切接触饲养人员、仔猪、育肥猪的喉头拭子、母猪阴道拭子及公猪的精液共183份。分别使用间接血凝诊断试剂盒、法国ELISA试剂盒检测抗体;利用直接荧光染色法检测抗原,包括166份猪喉头拭子、阴道拭子和精液以及17份饲养人员喉头拭子。结果发现间接血凝诊断试剂盒、ELISA试剂盒抗体阳性率分别为4.14%和2.17%;抗原平均阳性率为14.8%,其中精液阳性率达到37.5%,母猪阴道拭子阳性率为27.5%,饲养人员阳性率为23.5%。本试验证实北京郊区猪嗜流产衣原体在种公猪、母猪群和饲养员呈高流行趋势。同时研究结果显示,5个被调查的规模化猪场嗜流产衣原体抗体阳性率显著低于有报道的其他省市,这与间接血凝诊断试剂盒检测结果高于ELISA试剂有关。针对发现的问题,必须采取有效措施控制种公猪精液污染衣原体现象,阻断向母猪和饲养人员传播,以降低人兽共患病发生的风险。     

Winter dysentery in dairy herds: electron microscopic and serological evidence for an association with coronavirus infection.

Faeces and, or, paired sera were collected from cows in six dairy herds with classical winter dysentery. Similar samples were collected from cows in three other dairy herds experiencing non-haemorrhagic diarrhoea during the survey period. Coronavirus was the only enteric pathogen identified by immune electron microscopy (IEM) in all six outbreaks, occurring in 26 of 29 (90 per cent) of the affected cows and in one of 11 normal cows from the same herds. Nineteen of 26 affected cows (73 per cent) developed greater than four-fold increases in neutralising antibody titres to the Mebus strain of bovine coronavirus, compared with two of eight normal cows in the same herds. No cows showed greater than four-fold increases in antibody titres to bovine virus diarrhoea virus. None of the cows from the three herds with non-haemorrhagic diarrhoea shed coronavirus in faeces detectable by IEM or developed greater than two-fold rises in coronavirus antibody titres in paired sera. No enteric pathogens were identified in two of the herds. However, two cows in the third herd shed a group B rotavirus detected by IEM. These findings provide additional evidence for a possible role for bovine coronavirus in the aetiology of winter dysentery. Furthermore, this is the first report of a group B rotavirus associated with diarrhoea in adult cattle.     

The bovine immune response to Brucella abortus IV. Studies with a double immunodiffusion test for antibody against A2.

A double immunodiffusion test for precipitins against Brucella antigen A2 was developed and applied to a variety of samples. The A2 precipitins were produced by a heifer infected with B. abortus strain 2308, cattle vaccinated with killed B. melitensis strain H38 or live B. abortus strain 19 and by a dog infected with B. canis. Precipitins were also detected in the second International Standard for anti-Brucella abortus serum, in several anti-B. canis sera and at low levels in one anti-B. ovis serum tested. Antisera produced in calves against Yersinia enterocolitica serotype 0:9 had no anti-A2 activity despite titers greater than or equal to 1/1024 and greater than or equal to 1/80 in standard Brucella agglutination and CF tests, respectively. The test for A2 precipitins lacked specificity as weak reactions were obtained with five of 295 sera from brucellosis-free herds. This test was relatively insensitive, detecting precipitins in only 16 of 24 sera from infected cattle and 27 of 54 sera positive by complement fixation and enzyme labelled antiglobulin tests performed with whole cell and smooth lipopolysaccharide antigens, respectively. The A2 precipitins were detected in nine sera from five cattle, in two infected herds, which were negative by agglutination and complement fixation tests.     

Indirect fluorescence antibody studies of porcine cytomegalo virus infections in the Netherlands

Summary Sera from 683 pigs of 41 swine herds with clinical atrophic rhinitis (A R), from 477 pigs of 37 herds with no A R history, from 267 breeding sows and breeding boars for slaughtering, from 22 boars at an artifical insemination centre, and from 103 SPF pigs were tested for the presence of antibodies to porcine cytomegalo virus (PCMV). The herds examined were spread all over the Netherlands. For the presence of antibodies to PCM V the indirect fluorescence antibody test was used. To obtain the antigen, the PCMV had been grown in pig lung macrophage cultures in Petri dishes for 10-12 days. These macrophages were dropped into the wells of slides. The serum dilution 1:20 of all the 103 sera from SPF pigs were negative, but 93 per cent of the other sera were positive. No marked differences were found between swine herds with clinical atrophic rhinitis and herds with no A R history. The FA titres in both types of herds seem to be at a comparable level.     

Case-control study of factors associated with arthritis detected at slaughter in pigs from 49 farms

Data were collected on the housing, management and disease factors in the weaning and finishing units of 49 integrated pig herds, 24 of them with a high incidence of arthritis at slaughter (case herds) and 25 with a low incidence (control herds). A median of 5.2 per cent (range 3.7 to 12.4 per cent) of the slaughtered pigs in the case herds had arthritis at meat inspection, compared with 2.2 per cent (range 0.3 to 2.8 per cent) in the control herds. In the farrowing units, high clinical sign scores for the lactating sows and piglets less than one week old and a low age at castration were associated with the case herds. In the weaning units, the herds with open partitions between the pens were 5.6 times more likely to be a case herd than the herds with solid walls. A higher age at weaning and moving the piglets at weaning from the farrowing pen instead of the sows decreased the likelihood of being a case herd. In the finishing units, a higher score for clinical signs, using a proper hospital pen, disinfecting the pens between the groups and using a feeding plan increased the likelihood of being a case herd. In total, 145 condemned joints, a median of four (up to six per herd), were collected at the slaughterhouse. In the case herds, 71 of 76 joints (93.4 per cent) had lesions related to osteochondrosis and in the control herds 66 of 69 joints (95.6 per cent) had such lesions. Only two of 11 joints from the case herds and one of 12 joints from the control herds that were examined bacteriologically were positive for Stapylococcus aureus and/or Streptococcus species.     

Survey of campylobacter, salmonella and mycoplasmas in house crows (Corvus splendens) in Malaysia

House crows (Corvus splendens) in Selangor, Malaysia were examined for the presence of Campylobacter species, Salmonella species, Mycoplasma gallisepticum and Mycoplasma synoviae by serology, culture and pcr. For the detection of Campylobacter and Salmonella species swabs were taken either from the intestine or cloaca. For the detection of mycoplasmas, swabs were taken either from the choanal cleft or trachea for culture and pcr and serum samples were tested by the rapid serum agglutination (rsa) and monoclonal antibody-blocking elisa (mbelisa) for antibodies to M gallisepticum and M synoviae. For campylobacter, 25.3 per cent of the crows were positive by culture, and the species identified were Campylobacter jejuni and Campylobacter coli. No Salmonella species were isolated. Four of 24 swabs were positive for M gallisepticum dna but none gave positive results for M synoviae dna. No M gallisepticum or M synoviae antibodies were detected by rsa but 60 per cent of the sera gave positive reactions for M gallisepticum and 13 per cent gave positive reactions for M synoviae by mbelisa.     

Serological survey of encephalomyocarditis virus infection in pigs in France

Samples of serum from 76 gilts, 1440 sows, 1473 piglets and 3093 finishing pigs from 96 farrow-to-finish herds were tested for antibodies to encephalomyocarditis virus (EMCV) in microtitre serum neutralisation tests employing two strains of virus, one associated with myocarditis and the other with reproductive failure. The total seroprevalence of EMCV infection was 2.48 per cent. There was no significant difference between the seroprevalence of the reproductive failure strain (1.6 per cent) and the myocardial strain (1.85 per cent). The seroprevalence was higher in the gilts (6.57 per cent) and sows (5.13 per cent) than in the piglets (1 per cent) and finishing pigs (1.84 per cent), and the highest titres were observed in the sows (1:540) and finishing pigs (1:640). In the gilts, the difference in seroprevalence between the reproductive failure strain (3.95 per cent) and the myocardial strain (5.33 per cent) was wider than in the other groups.     

Prevalence of Chlamydia infections in breeding sows and their importance in reproductive failure

To investigate the prevalence of chlamydial infection and their significance for reproductive disorders in sow breeding herds in Germany, blood samples of 1493 pigs were taken for a serological survey by enzyme-linked-immunosorbent-assay (ELISA). Antibodies to Chlamydiae were found in 33% of the samples, in all herds investigated responders could be detected. The rate of seropositive animals in different farms varied from 4.3% to 72.7%. The percentage of positive responders in the farms correlated positively with the occurrence of MMA-syndrome (mastitis, metritis, agalactia), return to oestrus and diseases of the piglets. Also these herds showed less weaned piglets per sow and litter. Comparison of seronegative and seropositive sows within single farms revealed also worse reproductive data for seropositive sows. A significant relationship could be found between farms with a high quota of sero-positive sows and poor hygiene status as well as poultry keeping. As a second step 124 cervical swabs and 9 aborted piglets were investigated for chlamydial antigen by capture-ELISA and polymerase chain reaction (PCR). Using the capture-ELISA for investigation only 3 probes with chlamydial antigen could be detected, however, examination by the more sensible PCR gave positive results in 50% of the probes. 20% of the PCR-positive animals were clinically healthy sows, 80% of the PCR-positive probes originated from sows with reproductive disorders. A significant relationship could be shown between PCR-positive probes and the incidence of abortion and litters with stillborn piglets and piglets with low viability. Swabs from 93 of the 124 sows were investigated as well for other bacterial pathogens of reproductive disorders. A high degree of micro-organisms of different species could be detected in 70% of the samples of sows with reproductive disorders and in 35% of the samples of clinically healthy sows. Species differentiation of the chlamydial antigen positive samples was done by southern blot hybridisation. Herewith C. psittaci could be diagnosed in all positive samples. Additionally 8 probes revealed a mixed infection with C. psittaci and C. trachomatis. The results of the present study show, that the prevalence of chlamydial infections in breeding herds is high and underline the importance of chlamydial infections for reproductive disorders. Single chlamydial infections as well as mixed infections with other pathogens must be considered.     

Characterization of an atypical biotype of Brucella abortus.

Brucella abortus strains were isolated from bovine tissue and milk samples from seven Ontario herds. The isolates were characterized by colonial morphology, requirement of CO2 for growth, lysis by Tbilisi phage, biochemical tests and agglutination in monospecific sera. They resembled B. abortus biotype 2 (on the basis of sensitivity to thionin and basic fuchsin) and biotype 4 (on the basis of agglutination with anti-Brucella "M" but not anti-Brucella "A" absorbed sera). Sodium dodecyl sulphate-polyacrylamide gel electrophoresis of these isolates and B. abortus biotypes 1, 2 and 4 showed similar profiles. Immunoblots with anti-A and anti-M absorbed sera showed different antigenic regions reacting with the specific sera and also confirmed that the atypical B. abortus isolates were serologically similar to biotype 4.