Animal industry and veterinary science in eastern New Guinea: World War II to Independence, 1975.

We have described the efforts of the Australian administration of the Territory of Papua and New Guinea (TPNG) to establish a veterinary service and viable animal industries. These efforts began with planning before the end of World War II in 1945 and continued until independence in 1975. Whereas pre-war cattle had mostly been used to control grass on plantations, post-war, the objective was to use the country's extensive, unoccupied grasslands for cattle production. During this period, the cattle population increased from 4000 to more than 150,000. The greatest success was achieved in herds with crosses of Bos indicus and Bos taurus owned by expatriates. The only serious disease constraint on production was myiasis due to Chrysomya bezziana, a parasite throughout New Guinea and South-east Asia. Attempts were made to increase the productivity of the indigenous pig population. Success was limited by a failure to manage inadequate nutrition and internal parasites and to fully understand the cultural aspects of pig ownership. Similar problems inhibited chicken production in villages. The serious viral infections of pigs and birds were absent from TPNG, but a border with Indonesian Papua represents a potential route for the incursion of animal diseases exotic to both Papua New Guinea and to Australia.

Animal industry and veterinary science in eastern New Guinea 1: before World War II.

Until the latter part of the nineteenth century, there were no domestic animals other than pigs, dogs and poultry in the island of New Guinea. From 1889 onwards, occupying authorities, missionaries and settlers from Germany, the UK, Japan and Australia imported ruminants, pigs and horses. Some of these importations were from Asia. This paper describes some outcomes of those importations and the potential hazards for Australia entailed in them.

Comparison of footbathing and vaccination to control ovine footrot in an experimentally infected flock.

OBJECTIVE: Compare footbathing and vaccination for control of footrot during a transmission period in a sheep flock deliberately infected with multiple strains of Dichelobacter nodosus. METHODS: The strains included a known virulent strain, a benign strain and several intermediate strains. The resulting footrot was clinically intermediate. A total of 1450 Polwarth sheep aged 1-3 years were allocated to one of five treatment groups: untreated, weekly walkthrough zinc sulfate footbathing, 1-hour stand-in Footrite® footbathing every 3 weeks, vaccination with a commercial multivalent whole-cell vaccine and vaccination with a novel recombinant DNA fimbrial vaccine. There were four replicates, in four paddocks. RESULTS: Of the untreated animals, 76% had footrot. Footbathing, either weekly or every 3 weeks, restricted the prevalence to 6/283 (2%; 97% effective) and 18/275 (6.5%; 91% effective), respectively. This was significantly lower than the prevalence in either the untreated or vaccinated group (P < 0.001). Weekly footbathing resulted in significantly fewer affected sheep than footbathing for 1 h every 3 weeks (P < 0.05). Vaccination with either whole-cell or recombinant vaccines significantly (P < 0.001) reduced the prevalence ((142/280 (51%; 33% effective), 114/278 (41%; 46% effective) respectively), with the recombinant vaccine superior (P < 0.05) to whole-cell vaccination. Significantly (P < 0.05) fewer 1-year-old sheep had footrot than older sheep. A single Footrite treatment reduced the prevalence to 12% (53/445) compared with a prevalence of 57% (27/47) for untreated sheep (79% effective). CONCLUSION: In this study footbathing was more effective than vaccination at controlling and treating multistrain footrot.

Relationship between the likelihood of footrot elimination from a flock and the virulence of the strain of Dichelobacter nodosus present.

OBJECTIVE: To assess ability to eliminate different strains of footrot in sheep using inspection and culling of affected sheep. METHODS: A flock of 1417 Polwarth sheep that had deliberately been infected with seven different strains of Dichelobacter nodosus and undergone different control measures prior to eradication, including zinc sulfate footbathing and vaccination, were subjected to visual foot inspection on a number of occasions. Sheep identified as infected or having any foot abnormalities were removed from the flock at each inspection. The experiment had three replicates and a small number of untreated control animals. Sheep were examined following the inspections to assess the effectiveness of elimination and additional measures were implemented in two of the three replicates in an attempt to eliminate all strains of D. nodosus. RESULTS: Three strains of D. nodosus were apparently successfully eliminated from all replicates (strains A, E and H). Strains B and C were detected in one replicate each, despite additional inspections. The three stains that were eliminated were the more virulent strains and the two strains that remained were the least virulent. No assessment could be made on a further two strains. CONCLUSIONS: The application of a rigorous inspection and culling program resulted in the elimination of the more virulent D. nodosus strains, but did not result in the elimination of all D. nodosus strains on all occasions.

Artificial infection of sheep with multiple strains of Dichelobacter nodosus to induce footrot.

OBJECTIVES: To establish multiple strains of Dichelobacter nodosus in two flocks of sheep and to assess the virulence of five of these strains. METHODS: In experiment 1, sheep were challenged with five D. nodosus strains, which varied in both virulence and serotype. In experiment 2, consisting of four replicates (paddock groups), sheep were challenged with seven different strains of D. nodosus. In experiment 3, sheep were challenged with one of five D. nodosus strains. RESULTS: In experiment 1, at 28 days post challenge, four of the five challenge strains were present. Multiple-strain infections were present in 27 feet, with isolates from three serogroups being recovered from 5 feet, and four serogroups from 1 foot. Challenged hind feet were more frequently affected (P < 0.001). In experiment 2, four of the seven strains were recovered from one replicate and three strains from the remaining three replicates. Significantly more hind feet were affected (262/471, 55.6%) than front feet (198/481, 42%) (P < 0.001). Clinically, in both experiments 1 and 2 the footrot resembled an intermediate form, despite the inclusion of a virulent strain of D. nodosus. In experiment 3, this virulent strain caused a higher prevalence of more severe footrot, a greater mean total foot score and, in Merino sheep, resulted in significantly lower weight gains (P < 0.05). Interaction between D. nodosus strain and breed occurred, with Polwarth sheep being significantly more severely affected by one strain than Merino sheep. DISCUSSION: The clinical expression of multiple-strain infections has implications for both research and control of footrot. A novel method of control is proposed.

Effect on time in quarantine of the choice of program for eradication of footrot from 196 sheep flocks in southern New South Wales.

OBJECTIVE: To identify and compare programs for eradicating virulent footrot (VFR) chosen by owners of quarantined sheep flocks in southern New South Wales. METHOD: Data from 196 sheep flocks in the Wagga Wagga and Young Rural Lands Protection Boards were used to determine the program chosen, the influence of flock size on the program chosen and the effects of the program chosen and the use of contractors on the time in quarantine. RESULTS: The most popular programs in flocks using a single program were: total destocking (61/173; 35.3%) and inspection and culling of affected animals (71/173; 41.0%). Treatment of known infected animals was chosen in 41 flocks and of those, 10 (5.8%) used antibiotics for treatment and 31 (17.9%) used foot-bathing. Combined programs were used in 23 flocks and in 10 flocks a change of program occurred before eradication was achieved. The choice of program was, to some extent, affected by flock size, with owners of small flocks (<500 sheep) more likely to destock. The chosen program strongly influenced the time in quarantine, the shortest time being for destocking (mean 284 days), followed by culling of infected sheep (395 days), treatment with antibiotics (433 days) and finally foot-bathing (502 days). Time in quarantine was significantly shorter when contractors were used. CONCLUSION: All the options chosen led to the eradication of VFR. However, in this sample both the choice of program and the use of contractors influenced the time taken to achieve eradication and therefore the time in quarantine. Based on time in quarantine, foot-bathing was the least desirable option for the eradication of VFR because of the significantly greater time involved, perpetuation of risk to neighbours and increased cost of inspections. These findings were derived from flocks that were quarantined, but they are relevant to all flock owners considering eradication of VFR.

Meningitis, sepsis and endocarditis among workers occupationally exposed to pigs.

BACKGROUND: Workers exposed to pigs can develop meningitis, sepsis or endocarditis due to infection with Streptococcus suis transmitted from pigs to man. AIMS: To estimate the risk of these diseases. METHODS: We used the Occupational Hospitalization Register (OHR) which holds information about occupation and hospital treatments for all adults in Denmark. A dynamic population of male workers exposed to pigs was identified every year from 1995 to 2006 by occupational and industrial groups. First hospital treatment or death in the following year due to meningitis, sepsis or endocarditis was identified by ICD-10 codes from the OHR. By comparison with all other economically active men in Denmark, the standardized incidence ratio (SIR) was calculated for these diseases. RESULTS: Among those exposed, we observed 32 cases of meningitis, sepsis and endocarditis during 140,118 person-years. In the reference group, we observed 2680 cases during 15,209,394 person-years. The SIR of the exposed group was 1.35 (95% CI: 0.95-1.92). Among the 32 cases, 7 cases of meningitis and sepsis were specified as caused by infection with streptococci. The SIR for these seven cases was 2.4 (95% CI: 1.1-5.0). CONCLUSIONS: Our study did not find that workers exposed to pigs had an overall increased risk of developing meningitis, sepsis or endocarditis.

Distribution and prevalence of footrot in Bhutan.

The first cases of footrot in Bhutan were reported in sheep in 1990 at the National Sheep Breeding Centre (NSBC), which supplies breeding animals to village sheep flocks throughout Bhutan. Despite the presence of footrot at the Centre the distribution of apparently disease-free sheep continued. Cases of footrot were reported in village flocks soon after the disease was diagnosed at NSBC. A national survey was designed to establish the distribution and prevalence of footrot in Bhutan. This detected footrot in 19/94 village sheep flocks surveyed. The 19 affected flocks were distributed among nine different administrative districts whereas the villages selected were in 13 of a total of 16 sheep growing districts. The highest within-flock prevalences were among the seven flocks sampled in Bumthang district (mean 20.4%). The prevalence of the disease within flocks was generally much lower in other affected districts and in three districts a single affected animal was identified in the sample of 14 sheep examined in each village. Nationally, footrot prevalence was estimated to be 3.1% (95% CI 2.16-4.04%). There was a positive association between the receipt of animals from NSBC and the presence of footrot. The prevalence of the disease was higher in flocks with a migratory system of management than in those using a sedentary system. The relative risk of there being footrot in a migratory flock was nine-times higher than in a non-migratory flock. Only one strain of Dichelobacter nodosus (serogroup B) was identified among the 234 isolates obtained from the 19 affected flocks. Sheep with footrot healed quickly when treated with a vaccine made from this strain.

The use of an autogenous Dichelobacter nodosus vaccine to eliminate clinical signs of virulent footrot in a sheep flock in Bhutan.

An outbreak of virulent footrot was investigated in a flock of 605 Merino cross-bred sheep in Bhutan. Conventional control methods in the preceding eight years had reduced its prevalence from 36-79% in different components of the flock to about 15% overall. Only one serogroup (B) of Dichelobacter nodosus was identified among 40 isolates cultured from affected sheep. A vaccine prepared from this strain was used in a pilot trial to compare the response of 14 treated and 14 untreated sheep. All affected, vaccinated animals in this trial healed quickly and were protected against re-infection while additional cases developed among untreated sheep during a period favourable for the spread of footrot. The serogroup B vaccine was administered to the whole flock for two successive years. No other footrot treatment was given during these or subsequent years. The whole flock was examined three times, foot by foot, for two years and twice yearly for another two years. When vaccination began there were 88 affected sheep in the flock, an affected sheep being defined as an animal with a foot-score of 2 or greater in one or more feet. There were neither affected sheep in the flock 30 days after the first dose of vaccine nor were any identified in later inspections. Virulent footrot, originating from the farm under investigation, persisted in neighbouring village flocks during this period. It was concluded that whole flock specific D. nodosus vaccination made a major contribution to the elimination of all clinical signs of footrot from the flock of 605 sheep where the condition had previously persisted for 10 years.

Effect of climatic region on the clinical expression of footrot of lesser clinical severity (intermediate footrot) in sheep.

OBJECTIVE: To determine if the clinical classification of intermediate footrot (IFR) is changed to virulent footrot (VFR) by a transfer of the infected flock to a region where climatic conditions are more favourable for the transmission of the disease. DESIGN: Clinical examination of two groups of Merino wethers infected with IFR; one group of 309 in a region considered less favourable for footrot and another group of 343 at a second site considered more favourable. PROCEDURES: After characterising the form of footrot at the first site, infection was established at the second site by mixing 142 wethers from the first site with 201 unrelated wethers considered to be free of IFR and VFR. Observations of clinical characteristics were made over a 16 month period during which an outbreak of footrot occurred. Clinical assessments were made by inspecting every foot of every sheep at regular intervals and allocating a footscore. Evidence that the same clonal lines of D. nodosus were responsible for the footrot at both sites was provided by serotyping of isolates and using omp gene RFLP as a molecular epidemiological tool. RESULTS: The disease at the first site was classified as IFR because 7% of the sheep developed a maximum footscore (MFS) of 4, the most severe category, despite relatively low rates of transmission. When the outbreak occurred at the second site, which was more suitable for footrot transmission, the maximum proportion of the flock that developed a MFS of 4 was 3.6%, confirming the initial classification of IFR. CONCLUSIONS: When a flock infected with IFR was moved to a region where climatic conditions were more favourable for footrot transmission, the clinical classification of the disease remained the same in both the original flock and in sheep exposed to the infection for the first time.

Eradication of footrot of lesser clinical severity (intermediate footrot).

OBJECTIVE: To determine if intermediate footrot (IFR) can be eradicated from a flock of sheep by inspection and culling of cases during a non-transmission period and if prior antibiotic treatment or vaccination increases the likelihood of eradication. PROCEDURE: A replicated field experiment that compared the three eradication strategies was followed by an observational study of the best of these applied in a commercial flock of 3000 sheep. RESULTS: In the replicated experiment, IFR was eradicated either by inspection and culling alone, or when combined with vaccination. Eradication failed when the sheep were treated with parenteral antibiotics before inspection and culling during the non-transmission period. In the whole-flock program, eradication by repeated inspection and culling of footrot cases during the non-transmission period was successful and the flock remained free of infection 3 years later. CONCLUSIONS: IFR can be eradicated by inspection and culling but latent infections, which may persist undetected for at least 34 weeks, require surveillance inspections to be repeated during the non-transmission phase of the program. The use of parenteral antibiotics as an aid to the eradication of IFR is contraindicated.

Eradication of virulent footrot from sheep and goats in an endemic area of Nepal and an evaluation of specific vaccination.

Programmes based on the identification and treatment of cases and the culling of animals refractory to treatment had failed to eradicate virulent footrot from two districts in the western region of Nepal. From 1993 to 1996 vaccination against two endemic virulent strains of Dichelobacter nodosus was tested for its potential to contribute to the eradication of footrot from the region. Only sheep and goats which had been free of signs of footrot at three inspections at monthly intervals before their annual migration to alpine pastures were eligible for inclusion. From November 1992, the treatment of cases identified during inspections included the injection of specific vaccine. Successfully treated cases migrated with their flocks but were excluded from the vaccine trial. Non-responding cases were culled. Forty combined flocks of sheep and goats (approximately 9500 animals) were used initially to compare three vaccination regimens. Eleven flocks (sheep and goats) were treated with two doses of specific vaccine (group A), nine (sheep and goats) were treated with commercial vaccine followed by specific vaccine (group B) and 10 (sheep and goats) were treated with two doses of commercial vaccine (group C) in March to April 1993 before the annual migration; 10 flocks (sheep and goats) remained unvaccinated (group D). Only sheep and goats free of signs of footrot were allowed to migrate. Nevertheless, virulent footrot recurred in many flocks three months later. However, its prevalence was significantly lower in group A than in the other three groups combined. Groups A, B and C then received the specific vaccine before their migrations in 1994 to 1996; group D remained unvaccinated. The annual programme of inspection and identification and treatment of cases continued for seven years, but the vaccinations ceased after four years. There was no recurrence of virulent footrot after November 1993. After the first season the virulent strains of D nodosus used in the specific vaccine could no longer be isolated, although antigenically distinct, benign strains of the organism persisted in cases of benign footrot.

Serogroup specific single and multiplex PCR with pre-enrichment culture and immuno-magnetic bead capture for identifying strains of D. nodosus in sheep with footrot prior to vaccination.

The identification of Dichelobacter nodosus present in a flock is a prerequisite to specific (autogenous) vaccination. Current methods of identification of the serogroup present in a population requires that the organisms be isolated, identified visually in mixed culture on streak plates, subcultured to purify and subjected to antigenic analysis. This process takes at least 3 to 4 weeks. This study describes the development of a simple and rapid serogroup specific PCR test for D. nodosus. A common forward primer was designed from the conserved amino-terminal region of the fimbrial gene (fimA) and 9 (A-I) serogroup specific reverse primers were designed from the carboxy-terminal regions of fimA of the different serogroups. To verify the specificity within D. nodosus, each specific primer pair was tested in PCR against 18 serogroups/serotypes (prototypes) and found to be specific for all the serotypes within the homologous serogroups. Eighty four other bacterial strains, either commonly occurring in sheep or found in the environment of sheep, and including organisms related taxonomically to D. nodosus, were used to check the specificity of these assays. They were found to be specific for D. nodosus as none of the 84 bacterial stains reacted. These primers detected 1 pg of purified chromosomal DNA, or 50-100 cells of D. nodosus in crude lysates. Sensitivity was markedly improved when an immuno-magnetic capture was employed. Single tube multiplex PCRs were tested with different combinations of common forward primer and groups of 3, 4 or 5 reverse primers chosen so that amplicon size for each reaction product was different. These were able to amplify DNA of isolates from all the relevant serogroups included in the reactions. These tests were evaluated with samples taken directly from lesions of footrot, either directly or preceded by DNA purification, immuno-magnetic capture, enrichment broth culture and culture on hoof agar media. Of these methods only PCR on mixed colonies from 4-day-old cultures on 4% hoof agar media yielded results of practical value.

Diagnosis of footrot in goats: application of ELISA tests for response to antigens of Dichelobacter nodosus.

Goats are an important natural host for footrot and are infected with Dichelobacter nodosus that have virulence characteristics similar to those of sheep strains. However, the humoral response of goats to D. nodosus antigens and the possibility of a serological diagnosis of footrot in goats have not been studied. With the aim of evaluating a diagnostic ELISA test, we investigated the primary immune response of goats to experimental and natural infection, the memory response in recovered animals, and the transfer and persistence of colostral antibodies in kids. Footrot stimulated the goat's immune system and, as in sheep, under-running lesions were the primary stimulus for production of anti-D. nodosus antibodies. The immune response could be detected in ELISA using either fimbrial or outer membrane protein (KSCN) antigens of D. nodosus. Antibody titres resulting from infection declined quickly after recovery and reached pre-infection levels within 3-4 months. Previously affected animals, however, mounted a memory response when injected with purified D. nodosus antigens. Antibody levels attained after anamnestic challenge were correlated with the maximum levels attained during infection, and were therefore indicative of the infection status. Anti-D. nodosus antibodies were also transferred to kids via colostrum, but these antibodies did not persist and therefore were unlikely to interfere with the diagnostic ELISA after 3 months of age. Though these ELISA tests were highly specific, their sensitivity was rather low. Therefore, they are only suitable for a herd diagnosis of footrot in goats and are dependent on the development of advanced under-running infections in a proportion of affected goats.

The type IV fimbrial subunit gene (fimA) of Dichelobacter nodosus is essential for virulence, protease secretion, and natural competence.

Dichelobacter nodosus is the essential causative agent of footrot in sheep. The major D. nodosus-encoded virulence factors that have been implicated in the disease are type IV fimbriae and extracellular proteases. To examine the role of the fimbriae in virulence, allelic exchange was used to insertionally inactivate the fimA gene, which encodes the fimbrial subunit protein, from the virulent type G D. nodosus strain VCS1703A. Detailed analysis of two independently derived fimA mutants revealed that they no longer produced the fimbrial subunit protein or intact fimbriae and did not exhibit twitching motility. In addition, these mutants were no longer capable of undergoing natural transformation and did not secrete wild-type levels of extracellular proteases. These effects were not due to polar effects on the downstream fimB gene because insertionally inactivated fimB mutants were not defective in any of these phenotypic tests. Virulence testing of the mutants in a sheep pen trial conducted under controlled environmental conditions showed that the fimA mutants were avirulent, providing evidence that the fimA gene is an essential D. nodosus virulence gene. These studies represent the first time that molecular genetics has been used to determine the role of virulence genes in this slow growing anaerobic bacterium.

Pilus ELISA and an anamnestic test for the diagnosis of virulent ovine footrot and its application in a disease control program in Nepal.

The immunological memory (anamnestic) responses in sheep recovered from virulent footrot (VFR) can be aroused by subcutaneous injection of outer membrane protein (OMP) antigens of Dichelobacter nodosus. The magnitude of this response is directly correlated to the highest antibody response attained during infection and memory lasts at least a year after recovery from VFR. However, some older animals show non-specific responses to OMP antigens. In this study an evaluation of D. nodosus pilus antigen for the anamnestic diagnosis of footrot in sheep was undertaken. The results indicated that the primary and anamnestic responses to pilus were similar in character to OMP antigen but were highly specific. The sensitivity of the procedure for detection of sheep with a history of VFR was approximately 80%. A low proportion of sheep with mild lesions due to virulent strains of D. nodosus reacted to anamnestic challenge. Anamnestic challenge with 10 microg pilus was used in a VFR surveillance program in migratory sheep flocks in Nepal. Conventional diagnostic methods could not be applied during the disease transmission periods in these flocks because of their migration to alpine pastures far away from human habitation. The results supported clinical and bacteriological findings suggesting that virulent strains of D. nodosus have apparently been eliminated from these flocks in Nepal.

Characterization of hemolysin of Moraxella bovis using a hemolysis-neutralizing monoclonal antibody.

A concentrated bacterial culture supernatant from the hemolytic Moraxella bovis strain UQV 148NF was used to immunize mice and generate monoclonal antibodies (MAbs). One, MAb G3/D7, neutralized the hemolytic activity of M. bovis and recognized a 94-kDa protein by Western blot analysis in hemolytic M. bovis strains representing each of the different fimbrial serogroups. Exposure of corneal epithelial cells to M. bovis concentrated culture supernatants demonstrated a role for an exotoxin in the pathogenesis of infectious bovine keratoconjunctivitis, while neutralization of hemolytic and cytotoxic activities by MAb G3/D7 implies that these activities are related or have common epitopes. The action of M. bovis hemolysin was further characterized in sheep erythrocyte preparations with a binding step and Ca(2+) required for lysis to proceed, similar to the RTX family of bacterial exotoxins. Neutralization of lytic activity in vitro is evidence for the presence of M. bovis antigens, which may be capable of protecting cattle from the development of infectious bovine keratoconjunctivitis.

Transmission of virulent footrot between sheep and goats.

OBJECTIVE: To determine the infectivity of ovine and caprine strains of Dichelobacter nodosus for both sheep and goats. DESIGN: Pen experiments in which 20 sheep and 19 goats were challenged directly with the two strains, and transmission experiments on pasture, using donors infected by experimental challenge. RESULTS: Sheep and goat strains of D nodosus infected both animal species in experimental challenges. Animals so infected transmitted footrot to both sheep and goats on pasture plots. A significantly smaller proportion of goats than sheep was infected when challenged with either strain. The interval between exposure and development of footrot in goats was longer than in sheep when recipient animals were exposed to infected donors on pasture. The disease was less invasive in goats than in sheep. CONCLUSIONS: With the strains of D nodosus used there was no evidence of host specificity. Direct transmission of footrot can occur between sheep and goats in the same environment. There is a need to include goats in ovine footrot eradication programs and vice versa.

PCR-RFLP of outer membrane proteins gene of Dichelobacter nodosus: a new tool in the epidemiology of footrot.

Currently only phenotypic epidemiological markers, serogrouping and virulence testing of Dichelobacter nodosus, are available for investigating footrot outbreaks in small ruminants. These methods have limitations in tracing the source of infection. In this study, a genotypic marker, PCR-RFLP of outer membrane protein gene, was used to characterize D. nodosus. The technique was evaluated in a controlled experiment involving two strains of bacteria. PCR-RFLP was found to be highly specific in differentiating isolates obtained from recipient animals infected with different strains. Subsequently, this technique was used to characterize isolates obtained from field cases of footrot in Nepal. A total of 11 patterns was recognized among 66 Nepalese D. nodosus isolates representing four different serogroups. PCR-RFLP also discriminated isolates with similar phenotypic characteristics. However, all isolates which, phenotypically, were virulent were represented by only two patterns irrespective of their serogroups. It is suggested that PCR-RFLP described here could be a useful epidemiological marker in the study of footrot.

Identification and characterisation of serogroup M among Nepalese isolates of Dichelobacter nodosus, the transmitting agent of footrot in small ruminants.

One thousand and sixty three isolates of Dichelobacter nodosus cultured between 1992 and 1996 from cases of footrot in sheep and goats of migratory flocks of Nepal were characterised by agglutination test using prototype antisera of the Australian classification system. Of those, sixty six isolates could not be classified into any of the nine serogroups (A-I). This study was therefore undertaken to characterise these isolates. It was established that they were agglutinated by antiserum against serotype M of an alternative classification system. The distinct antigenic character of these isolates was further confirmed by DNA sequence analysis of the gene for the fimbrial subunit protein of two of them. At a molecular level, these isolates were closer to the prototype of serogroup F, VCS 1017. However, when compared with VCS 1017, the number of amino acid substitutions (28) in the fimbrial protein of these isolates was similar to that expected between isolates of different serogroups. Because these isolates are antigenically similar to 'serotype' M, but meet all the criteria to be classified into an independent serogroup, it is proposed that these isolates together with isolates previously classified as serotype M be classified as 'serogroup M'.