Environmental factors associated With Toxoplasma gondii Exposure in Neotropical Primates of Costa Rica.

The apicomplexan parasite Toxoplasma gondii (T. gondii) has been found in more than 350 species of homoeothermic vertebrates in diverse climates and geographic areas. In most animals, T. gondii produces mild or asymptomatic infection. However, acute and hyperacute toxoplasmosis is associated with high mortality rates observed in Neotropical primates (NP) in captivity. These primates are distributed in 20 countries across the Americas, and although infection has been reported in certain countries and species, toxoplasmosis in the wild and its impact on NP population survival is unknown. Differences among species in exposure rates and disease susceptibility may be due in part to differences in host behavior and ecology. Four species of NP are found in Costa Rica, i.e., howler (Alouatta palliata), spider (Ateles geoffroyi), capuchin (Cebus imitator), and squirrel monkeys (Saimiri oerstedii). This study reports NP exposure to T. gondii using the modified agglutination test in 245 serum samples of NP (198 wild and 47 from captivity) from Costa Rica. Associations of serostatus with environmental (forest cover, annual mean temperature), anthropogenic (human population density), and biological (sex) variables in howler and capuchin monkeys were evaluated. The seroprevalence among wild NP was 11.6% (95% CI = 7.7-17.34), compared with 60% in captive monkeys (95% CI = 44.27-73.63), with significant differences between species (X 2 = 20.072; df = 3, p = 0.000164), suggesting an effect of behavior and ecology. In general, antibody titers were low for wild NP (<1:128) and high for captive NP (>1:8192), suggesting higher exposure due to management factors and increased life span in captivity. Seropositivity in howler monkeys was positively related to forest cover and inversely related to annual rainfall. For capuchins, annual rainfall was inversely related to seropositivity. Surveillance of T. gondii exposure in NP in captivity and in the wild is required to understand drivers of the infection and develop novel strategies to protect them.

Brucellosis caused by the wood rat pathogen Brucella neotomae: two case reports.

BACKGROUND: Brucellosis is a chronic bacterial disease caused by members of the genus Brucella. Among the classical species stands Brucella neotomae, until now, a pathogen limited to wood rats. However, we have identified two brucellosis human cases caused by B. neotomae, demonstrating that this species has zoonotic potential. CASES PRESENTATION: Within almost 4 years of each other, a 64-year-old Costa Rican white Hispanic man and a 51-year-old Costa Rican white Hispanic man required medical care at public hospitals of Costa Rica. Their hematological and biochemical parameters were within normal limits. No adenopathies or visceral abnormalities were found. Both patients showed intermittent fever, disorientation, and general malaise and a positive Rose Bengal test compatible with Brucella infection. Blood and cerebrospinal fluid cultures rendered Gram-negative coccobacilli identified by genomic analysis as B. neotomae. After antibiotic treatment, the patients recovered with normal mental activities. CONCLUSIONS: This is the first report describing in detail the clinical disease caused by B. neotomae in two unrelated patients. In spite of previous claims, this bacterium keeps zoonotic potential. Proposals to generate vaccines by using B. neotomae as an immunogen must be reexamined and countries housing the natural reservoir must consider the zoonotic risk.

Diversity of Aerobic Bacteria Isolated from Oral and Cloacal Cavities from Free-Living Snakes Species in Costa Rica Rainforest.

Costa Rica has a significant number of snakebites per year and bacterial infections are often complications in these animal bites. Hereby, this study aims to identify, characterize, and report the diversity of the bacterial community in the oral and cloacal cavities of venomous and nonvenomous snakes found in wildlife in Costa Rica. The snakes where captured by casual encounter search between August and November of 2014 in the Quebrada González sector, in Braulio Carrillo National Park. A total of 120 swabs, oral and cloacal, were taken from 16 individuals of the Viperidae and Colubridae families. Samples were cultured on four different media at room temperature. Once isolated in pure culture, colonies were identified with the VITEK® 2C platform (bioMérieux). In order to test the identification provided on environmental isolates, molecular analyses were conducted on 27 isolates of different bacterial species. Specific 16S rDNA PCR-mediated amplification for bacterial taxonomy was performed, then sequenced, and compared with sequences of Ribosomal Database Project (RDP). From 90 bacterial isolates, 40 different bacterial species were identified from both oral and cloacal swabs. These results indicate the diversity of opportunistic pathogens present and their potential to generate infections and zoonosis in humans.

Epidemiology of bovine brucellosis in Costa Rica: Lessons learned from failures in the control of the disease.

Brucellosis, caused by Brucella abortus is a major disease of cattle and a zoonosis. In order to estimate the bovine brucellosis prevalence in Costa Rica (CR), a total 765 herds (13078 bovines) from six regions of CR were randomly sampled during 2012-2013. A non-random sample of 7907 herds (532199 bovines) of the six regions, arriving for diagnoses during 2014-2016 to the Costa Rican Animal Health Service was also studied. The prevalence estimated by Rose Bengal test (RBT) ranged from 10.5%-11.4%; alternatively, the prevalence estimated by testing the RBT positives in iELISA, ranged from 4.1%-6.0%, respectively. However, cattle in CR are not vaccinated with B. abortus S19 but with RB51 (vaccination coverage close to 11%), and under these conditions the RBT displays 99% specificity and 99% sensitivity. Therefore, the RBT herd depicted in the random analysis stands as a feasible assessment and then, the recommended value in case of planning an eradication program in CR. Studies of three decades reveled that bovine brucellosis prevalence has increased in CR. B. abortus was identified by biochemical and molecular studies as the etiological agent of bovine brucellosis. Multiple locus variable-number tandem repeat analysis-16 revealed four B. abortus clusters. Cluster one and three are intertwined with isolates from other countries, while clusters two and four have only representatives from CR. Cluster one is widely distributed in all regions of the country and may be the primary B. abortus source. The other clusters seem to be restricted to specific areas in CR. The implications of our findings, in relation to the control of the disease in CR, are critically discussed.

High Prevalence of Multidrug-Resistant Community-Acquired Methicillin-Resistant Staphylococcus aureus at the Largest Veterinary Teaching Hospital in Costa Rica.

Methicillin-resistant Staphylococcus aureus (MRSA) is a pathogen associated with severe infections in companion animals present in the community, and it is diagnosed in animals admitted to veterinary hospitals. However, reports that describe the circulation of MRSA in animal populations and veterinary settings in Latin America are scarce. Therefore, the objective of this study was to determine the prevalence and investigate the molecular epidemiology of MRSA in the environment of the largest veterinary teaching hospital in Costa Rica. Preselected contact surfaces were sampled twice within a 6-week period. Antimicrobial resistance, SCCmec type, Panton-Valentine leukocidin screening, USA type, and clonality were assessed in all recovered isolates. Overall, MRSA was isolated from 26.5% (27/102) of the surfaces sampled, with doors, desks, and examination tables most frequently contaminated. Molecular analysis demonstrated a variety of surfaces from different sections of the hospital contaminated by three highly related clones/pulsotypes. All, but one of the isolates were characterized as multidrug-resistant SCCmec type IV-USA700, a strain sporadically described in other countries and often classified as community acquired. The detection and frequency of this unique strain in this veterinary setting suggest Costa Rica has a distinctive MRSA ecology when compared with other countries/regions. The high level of environmental contamination highlights the necessity to establish and enforce standard cleaning and disinfection protocols to minimize further spread of this pathogen and reduce the risk of nosocomial and/or occupational transmission of MRSA.

Brucella canis is an intracellular pathogen that induces a lower proinflammatory response than smooth zoonotic counterparts.

Canine brucellosis caused by Brucella canis is a disease of dogs and a zoonotic risk. B. canis harbors most of the virulence determinants defined for the genus, but its pathogenic strategy remains unclear since it has not been demonstrated that this natural rough bacterium is an intracellular pathogen. Studies of B. canis outbreaks in kennel facilities indicated that infected dogs displaying clinical signs did not present hematological alterations. A virulent B. canis strain isolated from those outbreaks readily replicated in different organs of mice for a protracted period. However, the levels of tumor necrosis factor alpha, interleukin-6 (IL-6), and IL-12 in serum were close to background levels. Furthermore, B. canis induced lower levels of gamma interferon, less inflammation of the spleen, and a reduced number of granulomas in the liver in mice than did B. abortus. When the interaction of B. canis with cells was studied ex vivo, two patterns were observed, a predominant scattered cell-associated pattern of nonviable bacteria and an infrequent intracellular replicative pattern of viable bacteria in a perinuclear location. The second pattern, responsible for the increase in intracellular multiplication, was dependent on the type IV secretion system VirB and was seen only if the inoculum used for cell infections was in early exponential phase. Intracellular replicative B. canis followed an intracellular trafficking route undistinguishable from that of B. abortus. Although B. canis induces a lower proinflammatory response and has a stealthier replication cycle, it still displays the pathogenic properties of the genus and the ability to persist in infected organs based on the ability to multiply intracellularly.

Molecular Detection of Bartonella Species in Fleas Collected from Dogs and Cats from Costa Rica.

The bacterial genus Bartonella includes several species with zoonotic potential, some of which are common in domestic dogs and cats, as well as in their fleas. Because there is no previous information about the presence of Bartonella species in fleas from Central America, this study aimed at evaluating the presence of Bartonella spp. in fleas collected from dogs and cats in Costa Rica. A total 72 pools of Ctenocephalides felis and 21 pools of Pulex simulans were screened by conventional PCR to detect Bartonella DNA fragments of the citrate synthase (gltA) and the ß subunit RNA polymerase (rpoB) genes. Three (4.2%) pools of C. felis and five pools (22.7%) of P. simulans were found positive for Bartonella DNA. Sequences corresponding to Bartonella vinsonii subsp. berkhoffii strain Winnie, B. rochalimae, and an undescribed Bartonella sp. (clone BR10) were detected in flea pools from dogs, whereas Bartonella henselae and B. clarridgeiae sequences were identified in flea pools from cats. The detection of zoonotic Bartonella spp. in this study should increase the awareness to these flea-borne diseases among physicians and public health workers and highlight the importance of flea control in the region.

Resistencia antimicrobiana de cepas de Staphylococcus aureus, Costa Rica / Antimicrobial resistance of Staphylococcus aureus, Costa Rica

Determinar y comparar los perfiles de resistencia de cepas de S. aureus aisladas de quesos, producidos en la Zona Sur de Costa Rica y de un centro hospitalario de la misma región. Materiales y Métodos: Se analizaron 35 muestras de queso fresco, adquiridas durante los meses de setiembre y octubre del 2010 en la zona de San Vito de Coto Brus. A cada muestra se le realizaron recuentos de coliformes totales, coliformes fecales y Staphylococcus aureus. Adicionalmente se analizó presencia/ausencia de Listeria monocytogenes en 25 gramos del producto. A las cepas identificadas como S. aureus se les realizó la prueba de sensibilidad a los antibióticos mediante el sistema automatizado Vitek y la interpretación de los datos se realizó siguiendo las pautas del Clinical and Laboratory Standards Institute antimicrobial susceptibility testing 2011. Adicional a esto se recolectaron datos acerca de la sensibilidad de las cepas de S. aureus aisladas e identificadas en el Hospital de San Vito de Coto Brus en el mismo período. Resultados: El promedio obtenido para el recuento de coliformes totales fue de 9,7 X 10(6) UFC/g, para coliformes fecales de 6,7 X 10(5) y para S. aureus de 2,8 X 10(5) UFC/g, obteniéndose un 83 % de muestras positivas por esta bacteria. En cuanto a la resistencia antimicrobiana, se obtuvieron porcentajes de resistencia mayores en las cepas de origen clínico. Se encontró también que 23 de las cepas (96%) provenientes de muestras clínicas, presentaban resistencia a más de un antibiótico, mientras que siete de las obtenidas a partir de queso (27%) presentaban esta característica. Con respecto a los betalactamicos (ampicilina, oxacilina y penicilina) se observó la existencia de una diferencia estadísticamente significativa (p<0,01) entre las cepas de ambas fuentes, presentándose mayor resistencia en las de origen clínico. Conclusión: Ninguna de las cepas analizadas mostró resistencia a vancomicina, trimethoprim/sulfa ni linezolid...

Prevalencia de anticuerpos séricos contra la bacteria Brucella sp. en una población humana tropical

Prevalence of serum antibodies for the bacterium Brucella sp. in a tropical human population. Brucellosis is a systemic bacterial disease that can last years if not treated properly. The laboratory diagnosis is made by serological tests or isolation of the agent. We studied the prevalence of antibodies against Brucella sp. in inhabitants of rural communities of Cartago, Costa Rica, who are dedicated mainly to dairy cattle activities. We analyzed 714 representative samples with the Rose Bengal plate agglutination test (RBT) and the standard agglutination test in microplate (SAT). The overall prevalence was 0.87 %. No seroconversion was demonstrated for seropositive individuals. Even though 71 % of the population mentioned consuming non-pasteurized milk, no statistically significant association was found (1.09 %, IC 0.30-2.77) between milk consumption and the presence of antibodies suggestive of infection by Brucella sp. As a brucellosis high-risk population, the absence of symptoms in the population agrees with the observed titers of antibodies, suggesting a low previous exposure to the agent. Rev. Biol. Trop. 55 (2): 385-391. Epub 2007 June, 29.

La brucelosis es una enfermedad bacteriana generalizada con tendencia a la cronicidad si no se trata adecuadamente. El diagnóstico de laboratorio se realiza por el aislamiento e identificación del agente o en la mayoría de los casos mediante pruebas serológicas. En este estudio se informa la prevalencia de anticuerpos séricos contra Brucella sp. en habitantes de comunidades rurales de Cartago dedicadas principalmente a la ganadería de leche. Se analizó un total de 714 muestras representativas de la población mediante la técnica de aglutinación Rosa de Bengala en lámina (RBT) y la prueba de aglutinación en microplaca (SAT). Se encontró una prevalencia de 0.87 %, y en ninguna persona seropositiva se demostró seroconversión. Aún, cuando el 71 % de la población refirió consumir leche no pasteurizada, no se encontró asociación estadísticamente significativa (1.09 %, IC 0.30-2.77) entre el consumo de leche no pasteurizada y la presencia de anticuerpos sugestivos de infección por Brucella sp. A pesar de ser una zona de alta prevalencia de brucelosis bovina por Brucella abortus, la ausencia de síntomas en la población y los bajos títulos de anticuerpos observados sugieren una escasa exposición previa al agente.

Colonization of mutans streptococci in Costa Rican children from a high-risk population.

PURPOSE: The aim of the study was to determine the colonization of mutans streptococci (MS) in dental biofilm and saliva of children from a high-risk population with low socioeconomic status from San José, Costa Rica. METHODS: A total of 68 healthy babies from 8 to 20 months of age (mean age=13 months), with caries-free status and cariogenic feeding habits (eg, nursing bottle- or breast-feeding on demand) were examined. Children divided in two groups (<13 months and 14-20 months). Samples from dental biofilm and saliva from each subject were processed and plated in blood agar (BA) and Mitis-Salivarius-Bacitracin agar (MSB) to determine MS percentage. Statistical analysis included the analysis of variance test, and stratification by age of MS levels. RESULTS: Detectable MS levels in dental biofilm and saliva were found in 75% and 72% of children, respectively. Counts higher than 20000 colony forming units (CFU) per ml were found in 4% of saliva and in 35% of biofilm samples. The two groups showed significant differences in MS levels for dental biofilm (P=.036) and saliva (P=.009). Children older than 17 months presented higher MS levels (P<.05). Analysis by MS levels (<0.1%; 0.1-1.0%;>1.0% of MS) The two groups showed an insignificant association with increasing mean age. MS density was associated with mean number of erupted primary molars. CONCLUSIONS: Mutans streptococci colonization of dental biofilm and saliva from high caries risk infants is associated with age and dental development.

[Prevalence of serum antibodies for the bacterium Brucella sp. in a tropical human population]. / Prevalencia de anticuerpos séricos contra la bacteria Brucella sp. en una población humana tropical.

Prevalence of serum antibodies for the bacterium Brucella sp. in a tropical human population. Brucellosis is a systemic bacterial disease that can last years if not treated properly. The laboratory diagnosis is made by serological tests or isolation of the agent. We studied the prevalence of antibodies against Brucella sp. in inhabitants of rural communities of Cartago, Costa Rica, who are dedicated mainly to dairy cattle activities. We analyzed 714 representative samples with the Rose Bengal plate agglutination test (RBT) and the standard agglutination test in microplate (SAT). The overall prevalence was 0.87%. No seroconversion was demonstrated for seropositive individuals. Even though 71% of the population mentioned consuming non-pasteurized milk, no statistically significant association was found (1.09 %, IC 0.30-2.77) between milk consumption and the presence of antibodies suggestive of infection by Brucella sp. As a brucellosis high-risk population, the absence of symptoms in the population agrees with the observed titers of antibodies, suggesting a low previous exposure to the agent.