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We report a retrospective analysis of 1933 Brucella strains isolated from humans and animals in Latin American countries between 1968 and 1991 and in Argentina between 1994 and 2006. During the first period 50% of strains were from humans, mainly from Argentina, Mexico and Peru but, while B. suis was the main cause of infection in Argentina, B. melitensis was responsible for most infections in the other countries. In Argentina in the later years, B. melitensis and B. suis were observed more frequently than in the first period while isolation of B. abortus decreased. Of 145 B. melitensis human isolates, eight gave susceptibility patterns to dyes and penicillin and two were B. melitensis biovar 3, which has never been reported in animals. Forty-six percent of B. suis isolated were resistant to dyes which is an atypical feature in this species. [ABSTRACT FROM AUTHOR]

Ovine cattle was introduced into America during the Spanish conquest with the second journey of Columbus to the Antilles and was disseminated throughout the region. In 1587, sheep were introduced into Argentina, later developing into the "Creole" breed. We selected 486 animals from different Argentine provinces with the aim of determining the serological status of brucellosis caused by Brucella melitensis and Brucella ovis. For the detection of antibodies against smooth Brucella spp., the Rose Bengal test (RBT) was performed as screening test while the serum agglutination test (SAT) and 2 mercapto-ethanol (2ME) were run as a confirmatory technique. Moreover, for the detection of antibodies against rough Brucella spp., we used the rapid slide agglutination test (RSAT) for screening and an indirect ELISA (IELISA) as confirmatory assay. This study showed that the total positive percentage of brucellosis due to B. ovis was 2.9%. Excluding the animals mixed with the Suffolk breed; seropositivity would be 0.6%. All animals tested negative for brucellosis caused by B. melitensis., (Copyright © 2017 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.)

Consumption of inadequately pasteurized dairy products is the most common means of transmission of brucellosis. This report describes two foodborne outbreaks that occurred in families infected after consumption of fresh home-made cheese bought in different Argentine provinces. High resolution variable number of tandem repeats (VNTR)-based analysis revealed two well-defined groups comprising essentially identical profiles and corresponding to the two different outbreaks. Similar clinical findings in members of the same family could indicate that the differential virulence of different bacterial clones, as indicated by VNTR data, could have influenced the course of the disease. We observed the importance of adequate treatment in early stages of the disease; combination therapy and extended treatment for 6 weeks or longer yielded significantly better results. The risk of the foodborne transmission of this zoonotic disease and disease prevention should be considered.

The zoonotic risk of Brucella canis has been considered fairly high for persons who handle breeding dogs in kennels or are exposed to infected animals. Transmission to humans in other circumstances has been thought to be rare. We describe an uncommon outbreak of brucellosis caused by B. canis which, to the best of our knowledge, is the first reported in the literature. This outbreak involved six persons (three children and three adults), a bitch and three puppies which had close daily contact with the family. The clinical symptoms of the index case led to an erroneous diagnosis and the infection would have gone undiagnosed if culture had not been positive. This report aims to increase awareness of medical personnel of the need to order screening tests for children, immunodeficient persons or pregnant women presenting with fever of unknown origin, unexplained spleen or liver enlargement or other systemic signs. The emerging zoonotic potential of this disease in urban areas and the need to coordinate canine brucellosis surveillance systems should be evaluated.

There is little information in the literature on the clinical progress of brucellosis in patients affected by other non-infectious diseases; however, the infection can often trigger an exacerbation of existing underlying conditions in certain target organs. In this report we present four cases of brucellosis complicating previous diseases, and the difficulties in relation to their diagnosis and treatment. The study involved four patients with the following disorders: polycythaemia vera, pulmonary fibrosis, cirrhosis of the liver and arthritis of the knee. Brucellosis was diagnosed by classical serological and bacteriological methods. The strains involved could be isolated only in three of the four patients: two strains were Brucella abortus biovar 1 and one was Brucella suis biovar 1. Two patients relapsed 10 and 7 months after admission, another presented chronic brucellosis and received various therapy schemes, and one died. Since the best selection of antibiotics and the optimal duration of therapy remain unknown for patients having brucellosis complicated by previous pathologies, these remain at the discretion of the attending physician. Management of our patients was controversial in terms of the selection of antibiotics, duration of treatment and decision regarding surgery.

Brucella melitensis is highly pathogenic and constitutes a serious risk to public health. In Argentina, biovar 1 has been isolated from infected animals, but the Rev.1 strain vaccine is not authorised for use. This report describes nine atypical B. melitensis isolates obtained from humans. These isolates grew slowly, produced small colonies and were susceptible to penicillin and dyes, similar to the B. melitensis Rev.1 vaccine strain, but were inhibited by streptomycin 2.5 mg/L. The isolation of such atypical B. melitensis variants has never been reported from animals in Argentina, and could indicate the emergence of a new mutant variant.

Brucella canis is considered a rare cause of human brucellosis. The clinical importance of this infection may have been underestimated so far because of difficulties with presumptive diagnosis. The case described here presented symptoms compatible with brucellosis but the routine tests using Brucella abortus antigen were negative. The infection would have remained undiagnosed if culture had not been positive. This case illustrates the potential for a favourable outcome in Brucella canis diagnosis and supports recommendations for the use of B. canis serology. The infection should be suspected in patients with compatible symptoms and negative serology for B. abortus antigen.

The transmission of Brucella canis to man commonly occurs through contact with infected dogs or their secretions, or through direct laboratory exposure. The disease is underdiagnosed due to a general lack of serological testing facilities and misconceptions concerning its prevalence. This report shows the potential use of an indirect ELISA (IELISA) for the diagnosis of human brucellosis caused by B. canis in a population of patients negative by smooth-Brucella antigen tests but positive by rapid slide agglutination test (RSAT). One hundred and ten sera from asymptomatic people found negative by tests using smooth Brucella abortus antigen and by RSAT showed an IELISA specificity of 100 % when a cut-off value of 27 % positivity (%P) was selected. For 17 sera from patients with positive B. canis culture or in close contact with culture-positive dogs, the IELISA sensitivity was 100 % with the same cut-off value. The positive patients presented clinical symptoms similar to brucellosis caused by other species of Brucella and some of them received antibiotic treatment and made good progress. Using this cut-off value, we studied 35 patients with negative blood cultures but positive RSATs, and IELISA detected 18 as positive; of the 17 IELISA-negative, two were RSAT-positive at dilution 1 : 2 and 15 were weakly positive with pure serum. These samples were probably from patients at an early stage of infection or indicate false-positive results. No cross-reaction was observed among the sera from nine cases with a diagnosis other than brucellosis, but cross-reactivity was evident in sera from patients infected with smooth-Brucella species. Since routine brucellosis diagnosis does not include B. canis investigation, infection with this species may be more widespread than is currently suspected. The RSAT could be a suitable screening test for the diagnosis of B. canis human brucellosis, and a supplementary technique, such as IELISA, performed on all positive RSAT samples that were negative by B. abortus antigen could ensure diagnostic specificity and confirm the diagnosis.

Fluorescence polarization immunoassay (FPA) uses molecular rotational properties to measure antibody binding to antigen directly. The potential use of this method was assessed in comparison to a competitive enzyme immunoassay (CELISA) and conventional serological tests for the diagnosis of brucellosis on a total of 587 human sera. Based on 340 sera from asymptomatic blood donors with no evidence of brucellosis, the specificity of the FPA was 97.9 % using a cut-off value of 72 mP. Sera from Brucella-infected patients (11 Brucella melitensis, 32 Brucella abortus, 32 Brucella suis and one Brucella sp.) yielded a sensitivity estimate of 96.1 %. In tests on 84 sera from suspected brucellosis patients, the FPA detected 80 cases. Of 87 sera from patients with probable infection, 15 were detected by both CELISA and FPA, three by CELISA only and four by FPA only. The discrepancies in both groups involved sera with low, declining titres. The FPA uses a sample of 40 micro l serum, takes about 5 min to complete and has been demonstrated to be accurate for the detection of antibodies to B. abortus, B. melitensis and B. suis and for identifying patients suffering relapses. Because of the ease of the procedure, it could be readily adopted for use in clinical laboratories and blood banks.

The diagnosis of B. canis infection in dogs is based on bacteriological examination and serological methods including agglutination and gel diffusion tests. Bacteriological studies are the only methods that have been considered specific but, as intermittent periods of abacteraemia may occur, a negative blood culture cannot be used as a criterion for excluding canine brucellosis. Close contact between people and infected dogs increases the risk of transmission; however, its impact on public health is probably underestimated due to lack of reporting and inadequate diagnostic services. This paper describes an indirect enzyme-linked immunoassay (IELISA) procedure for the diagnosis of brucellosis caused by B. canis in a population of normal and infected dogs previously screened by the buffered plate antigen test (BPAT) and rapid slide agglutination test (RSAT). The serological survey was performed with 446 field sera. The 270 sera from the asymptomatic group found negative by BPA, RSAT and blood culture showed IELISA specificities of 96.7% and 100%, respectively, when cut-off values of OD 0.237 and 0.281 were selected. For 52 sera from culture-positive dogs, IELISA sensitivity was 100% with cut-off values of OD414 0.237 and 0.281. OD414 0.281 was selected because this value provided the highest accuracy with minimal false-negative and false-positive results. This cut-off value was used to study 124 blood culture-negative but RSAT positive sera. IELISA produced 107 positive results; the 17 sera that were negative by IELISA presented a wide range of reactivities by RSAT (2 were RSAT positive at 1 in 2 dilution and 15 were weakly positive with pure serum). These samples were probably from animals at an early stage of the infection or were false-positive results. The IELISA described here detects IgG and IgA antibodies that are useful for evaluating the clinical status of dogs. Although RSAT is a practical screening test, a supplementary technique such as IELISA should be used on all positive RSAT samples to ensure diagnostic specificity. Furthermore, people in contact with infected dogs could be investigated for possible transmission. The procedure described in this study was relatively simple and could have widespread applications.

The methods commonly used for human brucellosis serological testing are agglutination tests and the complement fixation test (CFT). Among the newer serological tests, primary binding assays were developed to improve sensitivity and specificity. The competitive enzyme immunoassay (CELISA) for the detection of serum antibody to Brucella is a multispecies assay which appears to be capable of differentiating vaccinal and cross-reacting antibodies from antibodies elicited by field infection in cattle. The competing monoclonal antibody used in this assay is specific for a common epitope of smooth lipopolysaccharide (S-LPS). In this study, we compared the CELISA to the classical tests for the diagnosis of human brucellosis. The CELISA cutoff value was determined to calculate its diagnostic specificity and sensitivity. A survey was performed with 911 sera. Of the sera, 341 were from an asymptomatic population that tested negative with conventional serological tests (screening and confirmatory). Based on these samples, the CELISA specificities were determined to be 99.7 and 100% with cutoff values of 28 and 30% inhibition (%I), respectively. In a further study with 393 additional sera from an asymptomatic population found negative by the conventional screening tests, the CELISA specificities were calculated to be 96.5 and 98.8% with cutoff values of 28 and 30%I. The CELISA sensitivities were determined to be 98.3 and 94.8% with cutoff values of 28 and 30%I, respectively, for sera from 116 individuals found positive by the classical tests. For the 51 culture-positive patients, CELISA was positive for 100%, the CFT was positive for 92%, and the standard tube agglutination test (TAT) was positive for 100%. The CELISA specificity was 100% for 31 sera from patients found negative by conventional serological tests but with brucellosis-like symptoms. The CELISA is fairly rapid to perform, somewhat faster than TAT, and cross-reacts less with other antigens (or antibodies) than the conventional tests. Further, the CELISA is simpler to perform that the CFT and may readily be standardized by the use of purified S-LPS antigen and monoclonal antibody for competition.

Brucellosis in Argentina is currently investigated in bank donor blood by the standard plate agglutination test (PAT). This study evaluated the buffered plate antigen test (BPA), now used to screen for bovine brucellosis, as a screen for human disease. Of 57 sera from patients with culture-confirmed brucellosis, 100% were detected with the BPA. Of 142 sera positive by rose bengal (RB) and complement fixation (CF), from patients with clinical evidence of brucellosis, the BPA detected 100%. Of 307 sera from a nonsymptomatic population that were RB and CF negative, the BPA detected 99.67% of the negative sera. The data indicate that the BPA is satisfactory compared to the other agglutination tests employed. It is an inexpensive and practical screening test and reduces the nonspecific reactions detected by the PAT.