Higher diversity of Brucella canis in Latin America, according to an MLVA_13 Bc analysis.

Brucella canis is the main causative agent of canine brucellosis, which affects domestic and wild canids and leads to clinical signs and symptoms of the reproductive and locomotor systems. Owing to the scarce information on this pathogen, here we addressed the genetic diversity of the circulating strains of this species in Argentina by following an MVLA_13 Bc scheme. The analyzed sample set consisted of 101 strains of B. canis isolates collected between 2006 and 2020 from canines of the Autonomous City of Buenos Aires (CABA) and other regions of Argentina, as well as 235 isolates from North America. The analysis yielded 336 variants (Hunter-Gaston Diversity Index, HGDI equal to 1.0) showing high diversity on a global scale. The analysis of the six most variable markers also reveled high diversity and allowed further analysis regarding variant relationships. Although the diversity obtained using both schemes (all or the 6 most variable markers) was higher for the Latin American than for the North American strains, we cannot discard that this was due to biases in the sampling methodology or to the different health policies employed in these regions regarding the management of infected individuals. Altogether, the Argentine circulating strains are genetically diverse, but with no apparent geographical association. The markers used in the MLVA_13 Bc are variable and highly useful for the evaluation of outbreaks. Furthermore, the reduced panel of 6 markers (MLVA_6 Bc) proposed in this study is convenient for the study of B. canis strain diversity.

Brucella canis Group 2 isolated in Argentina.

The aim of this study was to estimate the diversity and prevalence of both groups of Brucella canis 1 and 2 with and without deletion respectively in different areas of Argentina. A total of 104 bacterial cultures were typed as B. canis strains using the classical biotyping method. Two PCR assays were performed to confirm that all isolates were B. canis and not Brucella suis. The differentiation between groups 1 and 2 was achieved using another PCR assay and the diversity of B. canis isolates was assessed with four MLVA_16 markers. All strains belonged to Group 2. Bruce 09 marker (MLVA_16 assay) showed the greatest diversity. Only Group 2 of B. canis was identified among the strains evaluated. The markers chosen from the MLVA_16 allowed us to detect genetic diversity among the strains of B. canis studied.

Scavenger birds exploiting rubbish dumps: Pathogens at the gates.

Different bacteria are present in rubbish dumps used as food resources by various bird species. Birds may be good indicators of the presence of zoonotic diseases in these sites since they can be infected with zoonotic pathogens by foraging on organic waste, and can also act as carriers. We studied if foraging in rubbish dumps increases the occurrence of Salmonella spp. and Chlamydia psittaci in American black vultures (Coragyps atratus, hereafter black vultures) from northwest Patagonia. We compared these pathogens isolated from or detected in cloacae and oropharynx swabs in two different groups of black vultures: individuals trapped in (a) the Patagonian wild steppe and (b) in a rubbish dump. We found that black vultures are colonized by Salmonella spp. (particularly Salmonella enterica serotypes Typhi, Paratyphi A, Salmonella enterica subsp. arizonae) and Chlamydia psittaci. Interestingly, there were differences in the prevalence of Salmonella spp., especially Salmonella Typhi, between individuals foraging in the rubbish dump and the steppe, but not in the prevalence of Chlamydia psittaci. The pathogens isolated from black vultures may impact their health status but could also have health impacts in other bird species and even humans. In fact, Salmonella Typhi can cause severe disease in humans leading to death. Our results are globally relevant given that bacterial infections from rubbish dumps may affect different species exploiting these sites around the world. There is a need to better control pathogens in rubbish dumps to avoid the risk of infecting wildlife, which could act as potential dispersers and reservoirs of these pathogens.

Comparison of four polymerase chain reaction assays for the detection of Brucella spp. in clinical samples from dogs.

AIM: This study aimed to compare the sensitivity (S), specificity (Sp), and positive likelihood ratios (LR+) of four polymerase chain reaction (PCR) assays for the detection of Brucella spp. in dog's clinical samples. MATERIALS AND METHODS: A total of 595 samples of whole blood, urine, and genital fluids were evaluated between October 2014 and November 2016. To compare PCR assays, the gold standard was defined using a combination of different serological and microbiological test. Bacterial isolation from urine and blood cultures was carried out. Serological methods such as rapid slide agglutination test, indirect enzyme-linked immunosorbent assay, agar gel immunodiffusion test, and buffered plate antigen test were performed. Four genes were evaluated: (i) The gene coding for the BCSP31 protein, (ii) the ribosomal gene coding for the 16S-23S intergenic spacer region, (iii) the gene coding for porins omp2a/omp2b, and (iv) the gene coding for the insertion sequence IS711. RESULTS: The results obtained were as follows: (1) For the primers that amplify the gene coding for the BCSP31 protein: S: 45.64% (confidence interval [CI] 39.81-51.46), Sp: 95.62% (CI 93.13-98.12), and LR+: 10.43 (CI 6.04-18); (2) for the primers that amplify the ribosomal gene of the 16S-23S rDNA intergenic spacer region: S: 69.80% (CI 64.42-75.18), Sp: 95.62 % (CI 93.13-98.12), and LR+: 11.52 (CI 7.31-18.13); (3) for the primers that amplify the omp2a and omp2b genes: S: 39.26% (CI 33.55-44.97), Sp: 97.31% (CI 95.30-99.32), and LR+ 14.58 (CI 7.25-29.29); and (4) for the primers that amplify the insertion sequence IS711: S: 22.82% (CI 17.89 - 27.75), Sp: 99.66% (CI 98.84-100), and LR+ 67.77 (CI 9.47-484.89). CONCLUSION: We concluded that the gene coding for the 16S-23S rDNA intergenic spacer region was the one that best detected Brucella spp. in canine clinical samples.

Molecular detection of Neorickettsia risticii in Brazilian free-tailed bats (Tadarida brasiliensis) from Buenos Aires, Argentina

Neorickettsia risticii is the causative agent of Potomac Horse Fever, a severe febrile disease affecting horses, transmitted by trematodes species with a complex life cycle. A total of 30 insectivorous bats (Brazilian free-tailed bat Tadarida brasiliensis) were analyzed by PCR for presence of genus Anaplasma, Ehrlichia, Neorickettsia and Rickettsia. Three samples showed positive reactions for genus Anaplasma, Ehrlichia and Neorickettsia, and the sequences were 99.67% identical to Neorickettsia risticii. The role of bats in the life cycle of N. risticii has yet to be elucidated; however bats may be reservoirs for this bacterium. To our knowledge, this is the first evidence of N. risticii in Argentina.

Brucella canis causing infection in an HIV-infected patient.

From the blood culture of an HIV-positive patient with a febrile syndrome (CD4 count 385 cells/microL and viral load nondetectable), Brucella canis was isolated. The patient was presumptively infected from his dogs, which tested positive, and showed good outcome after the therapy with doxycycline-ciprofloxacin, and the HIV infection would seem not to have been influenced by brucellosis. To our knowledge, no other case of B. canis in the setting of HIV infection has been reported in the literature, and the emerging zoonotic potential of the disease in urban areas should be considered.

[Canine brucellosis in dogs in the city of Buenos Aires]. / Brucelosis canina en perros de la ciudad de Buenos Aires.

Canine brucellosis, caused by Brucella canis, provokes epidydimitis, testicular atrophy and sterility in male dogs, while in females the major symptom is miscarriage. Transmission to humans may be through contact with semen, urine and/or aborted fetuses of infected animals. Our study, observational and cross-sectional, focused on dogs in lower class neighborhoods and slums with a high rate of unmet basic needs (UBN) in 8 areas of the city of Buenos Aires. We studied 219 dogs: 184 females and 35 males, that tested negative to the buffered plate antigen test (BPAT), which ruled out infection with smooth species of Brucella. We detected anti-B. canis antibodies in 16 dogs (7.3%): 9 females and 7 males, relevant data since the urine of males is considered one of the vectors for the spread of the infection. Although we could run blood cultures on only 175 animals, we isolated B. canis in 3 (2 females and 1 male). Only 3 of the owners of dogs that tested positive consented to a serological diagnosis and two of them were positive. We highlight that the agar gel immunodiffusion test (IGID) proved to have low sensitivity, having detected only one of the 16 positive cases and none of the three confirmed by isolation. We conclude that in the areas studied, the detection of serologically positive dogs and the isolation of B. canis in 3 cases are indicators of the health hazard for the population exposed to it.

Brucelosis canina en perros de la ciudad de Buenos Aires / Canine brucellosis in dogs in the city of Buenos Aires

La brucelosis canina, causada por Brucella canis, provoca epididimitis, atrofia testicular y esterilidad en los perros, mientras que en las hembras el síntoma principal es el aborto. La transmisión al hombre puede ser por contacto con el semen, orina, y/o fetos abortados de animales infectados. El presente estudio de tipo observacional de corte transversal, se realizó en caninos de barrios y asentamientos con alto índice de necesidades básicas insatisfechas (NBI) en 8 áreas de la ciudad de Buenos Aires. Se estudiaron 219 perros, 184 hembras y 35 machos, que fueron negativos a la prueba de aglutinación con antígeno tamponado (BPAT), que descartó la infección con especies lisas del género Brucella. Sedetectaron anticuerpos anti-B. canis en 16 perros (7.3%), 9 hembras y 7 machos, este último dato es relevante ya que la orina de los machos es considerada uno de los medios de diseminación de la infección. Aunque sólo se pudieron tomar hemocultivos a 175 animales, en 3 (2 hembras y un macho) se aislaron B. canis. Sólo 3 de los dueños de los perros positivos accedieron al diagnóstico serológico y dos resultaron positivos. Destacamos que la prueba de inmunodifusión en gel de agar (IGDA) ha demostrado ser poco sensible, detectó sólo uno de los 16 casos positivos y ninguno de los tres confirmados por aislamiento. Concluimos que en las áreas estudiadas el hallazgo de perros serológicamente positivos y el aislamiento de B. canis en 3 casos, son indicadores del riesgo en el que se encuentra la salud de la población expuesta.

Canine brucellosis, caused by Brucella canis, provokes epidydimitis, testicular atrophy and sterility in male dogs, while in females the major symptom is miscarriage. Transmission to humans may be through contact with semen, urine and/or aborted fetuses of infected animals. Our study, observational and cross-sectional, focused on dogs in lower class neighborhoods and slums with a high rate of unmet basic needs (UBN) in 8 areas of the city of Buenos Aires. We studied 219 dogs: 184 females and 35 males, that tested negative to the buffered plate antigen test (BPAT), which ruled out infection with smooth species of Brucella. We detected anti-B. canis antibodies in 16 dogs (7.3%): 9 females and 7 males, relevant data since the urine of males is considered one of the vectors for the spread of the infection. Although we could run blood cultures on only 175 animals, we isolated B. canis in 3 (2 females and 1 male). Only 3 of the owners of dogs that tested positive consented to a serological diagnosis and two of them were positive. We highlight that the agar gel immunodiffusion test (IGID) proved to have low sensitivity, having detected only one of the 16 positive cases and none of the three confirmed by isolation. We conclude that in the areas studied, the detection of serologically positive dogs and the isolation of B. canis in 3 cases are indicators of the health hazard for the population exposed to it.