[Detection of toxigenic genes nheA, nheB and nheC in Bacillus cereus strains isolated from powdered milk samples in Costa Rica]. / Presencia de los genes de toxigenicidad nheA, nheBy nheC en cepas de Bacillus cereus aisladas de leches deshidratadas en Costa Rica.

Powdered milk is a frequently consumed product that does not need to be kept under cold conditions. Nevertheless, different microorganisms may contaminate it. Powdered milk is a highly consumed product by Costa Rican population, and Bacillus cereus is a potentially pathogenic bacteria associated to it, with the ability to develop toxins depending on the presence of the respective codifying genes. The aim of this study was to determine the presence of the toxigenic genes nheA, nheB and nheC from B. cereus strains, found in powdered milk sold at the Costa Rican national market. Five different lots of ten brands of powdered milk, distributed in the metropolitan area of San José, Costa Rica were analyzed. B cereus load was quantified using the Most Probable Number technique and identified using the Vitek system. The presence of the toxigenic genes was determined using the PCR technique. The isolation frequency of this bacteria in the powdered milk samples analyzed reached 50%, with populations ranging from 3 to > 100 MPN/g. Five out from nineteen strains were found positive for the three toxigenic genes, indicating contamination with potentially toxigenic B. cereus in powdered milk distributed in the national market, and an important risk for public. health.

[Arcobacter: a foodborne emerging pathogen]. / Arcobacter: un patógeno emergente de origen alimentario.

In the last three decades, several emergent diseases affecting human beings have been identified, most of them from infectious origin including bacterial, viral, parasitic and even difficult to classify as spongiform encephalopathy. Most of these are zoonotic as it is the case of Arcobacter, currently considered as an emerging and food borne pathogen, of growing importance for public health. The increase in the prevalence and incidence of cases associated to this bacteria as well as in the number of actual researches and reports, suggest that the infection in human beings and animals has been underestimated due to a lack in knowledge about this bacteria and of a standardized isolation protocols, as well as the use of correct identification methods and techniques. Increasing trends in the isolation of Arcobacter from animal derivates used as food and from samples taken during production processes, cause an augment in public health awareness, since there is little knowledge about the pathogenic potential of Arcobacter species and the few focused in this bacterial group, show many different transmission routes and host species. Given this, the objective of the present review is to actualize the reader in the most important characteristics of this bacterium, including its morphology, distribution, classification, transmission, association with water, food, pets and animals, as well as the laboratory isolation techniques, virulence factors and their antibiotic susceptibility patterns.

[Analysis of the microbiological quality and potential presence of Listeria monocytogenes in custard apple (Annona muricata), mango (Mangifera indica) and passion fruit (Passiflora edulis) pulps from Costa Rica]. / Análisis de la calidad microbiológica y potencial presencia de Listeria monocytogenes en pulpas de guanábana (Annona muricata), mango (Mangifera indica) y maracuyá (Passiflora edulis) costarricenses.

The objective of this work was to determine some of the indicators associated to shelf life, hygiene, process and storage conditions for some of custard apple, mango and passion fruit pulps distributed by the main supermarket chains of the Metropolitan Area of San José, Costa Rica, as well as to examine the potential presence of Listeria monocytogenes in them. Sixty fruit pulp samples were analyzed. Tests included pH determination, total aerobic plate count, yeasts and mold count, lactic bacteria count, total and fecal most probable number and the presence/absence of Listeria monocytogenes in 25 g of the product. Fruit pulp's pH ranged between 3,1 and 3,9, and the microbiological counts obtained were relatively low except for one industry. None of the samples analyzed presented total or fecal coliforms. The presence of Listeria monocytogenes was confirmed in three samples, all of them coming from industry C. Low microbiological counts obtained may be due to the addition of preserving substances and to the pasteurization of some of the products; lack of these two elements may allow the presence of dangerous bacteria such as Listeria monocytogenes.

[Determination of the antimicrobial capacity of green tea (Camellia sinensis) against the potentially pathogenic microorganisms Escherichia coli, Salmonella enterica, Staphylococcus aureus, Listeria monocytogenes, Candida albicans and Aspergillus niger]. / Determinación de la capacidad antimicrobiana del té verde (Camellia sinensis) contra los agentes potencialmente patógenos Escherichia coli, Salmonella enterica, Staphylococcus aureus, Listeria monocytogenes, Candida albicans y Aspergillus niger.

Many studies can be found in scientific literature demonstrating the antimicrobial capacity of different herbs, including green tea. Never-theless, many results are divergent or cannot be compared. Several green tea formulations may be found in market, but there is scarce or non-information about its activity. In this work, the potential antimicrobial effect of 50 samples of dry green tea and in 10% infusion against Escherichia coli, Salmonella enterica, Listeria monocytogenes, Staphylococcus aureus, Candida albicans and Aspergillus niger distributed in the metropolitan area of Costa Rica, was determined. This activity was compared with the effect produced by Chinese origin green tea (Camellia sinensis). Different solvents were evaluated for preparing polyphenol enriched extracts from green tea samples. Total phenols were determined using the Folin-Ciocalteu spectrophotometric methodology, using galic acid as reference. Antimicrobial activity of green tea extracts and infusions was evaluated using the microplate methodology described by Breuking (2006). Ethanol was the most efficient solvent used for the polyphenol extractions. There was no antimicrobial effect of the different green tea extracts and infusions against the microorganisms evaluated, except for Listeria monocytogenes, where the extracts of 70% of samples analyzed and the control showed an inhibitory effect in the 10.5 mg/mL and 1.05 mg/L concentrations. None of the infusions tested, including the control, showed any effect against this bacteria.

[Bacteriological quality and toxigenic Bacillus cereus detection in cooked white rice sold at the metropolitan area of San José, Costa Rica]. / Calidad bacteriológica y detección de Bacillus cereus toxigénicos en arroz blanco cocido expendido en el área metropolitana de la provincia de San José, Costa Rica.

The wide use of rice is one of the factors that favors its implication in food borne diseases, and one of the most important pathogens associated to it is Bacillus cereus. The aim of this work was to evaluate the microbiological quality of 50 samples of white cooked rice sold in restaurants at the Metropolitan Area of San José, Costa Rica, including the determination of the total aerobic plate count, the Most Probable Number of total and fecal coliforms and Escherichia coli. MPN of Bacillus cereus and the detection of nheA, nheB and nHeC genes, associated to its toxicity, was also performed. Procedures described in the Compendium of Methods for the Microbiological Examination of Foods were followed for the bacteriological analysis, multiplex PCR was used for the detection of genes following the methodology described by Hansen et al, 2001. 46% of the samples analysed were positive for total coliforms, 34% for fecal coliforms, 16% for E. coli and 10% for B. cereus, being 8% toxigenic. These facts suggest that white cooked rice may represent a risk for Pubic Health and that improvements shall be performed in order to offer a safe and high quality product to consumers.

Arcobacter butzleri: first isolation report from chicken carcasses in costa rica.

Arcobacter butzleri isolation from chicken carcasses in Costa Rica is reported for the first time. The isolated strains (P and R) were presumptively identified by their phenotypic characteristics. Definitive identification was made using a multiplex PCR assay for the simultaneous detection and identification of Arcobacter butzleri, Arcobacter cryaerophilus and Arcobacter skirrowii. These first isolations indicate the necessity of further investigation about the prevalence, distribution, ecology and interactions with human beings of this and other Arcobacter species.

[Bacteriological quality of traditional, organic and hydroponic cultured lettuce in Costa Rica]. / Comparación de la calidad bacteriológica de la lechuga (Lactuca sativa) producida en Costa Rica mediante cultivo tradicional, orgánico o hidropónico.

The main objective of this work was to evaluate the microbiological quality of lettuces commercialized in the Metropolitan Area of San José, Costa Rica, and cultured in different ways, in order to detect differences between the culturing methods and the risk that these products may represent for Public Health. The study was done at the Food Microbiology Laboratory, Universidad de Costa Rica, from March to July, 2010. 30 lettuce samples were analyzed (10 obtained by traditional culture, 10 by organic culture and 10 by hydropony). All samples were obtained from markets where their origin was certified. Total aerobic plate count, total and fecal coliforms count and Escherichia coli were determined to all samples, as well as the presence/abscense of Salmonella spp. and Listeria monocytogenes in 25 g. Results obtained show that there is no statistically significant difference (p < 0.001) between the different types of cultures analyzed for any of the parameters evaluated. An important percentage of the samples presented coliforms, nevertheless, just one E. coli strain was isolated from a traditionally cultured lettuce sample. Four different Salmonella spp. strains were isolated from the samples as well as one Listeria monocytogenes strain. Data obtained show that the consumption of this product, raw or without an adequate hygiene and disinfection may represent a risk for health. Also, from the bacteriological point of view, there is no significant difference between the culturing methods evaluated, suggesting that the specific directions for each type of culture are not followed or that there is an inadequate handling of the products or post harvest contamination.

[Evaluation of the polymerase chain reaction (PCR) for the detection and identification of Listeria monocytogens in fresh cheese coming from the metropolitan area of San José, Costa Rica]. / Evaluación de la prueba de reacción en cadena de la polimerasa (PCR) en la detección e identificación de Listeria monocytogenes en queso fresco proveniente del Area Metropolitana de San José, Costa Rica.

Food borne diseases are very important worldwide and their frequency is still high despite the different efforts focused in diminishing their morbidity and mortality. Listeria monocytogenes is one of the agents associated in this kind of diseases. In the lactic industry, this bacteria is important since raw milks as well as dairy products have been associated in outbreaks, being fresh cheese one of the most vulnerable products to the contamination with this bacteria. The traditional identification of the bacteria is done by a laborious, time consuming and low sensitive technique and the polymerase chain reaction may allow more precise and exact results in shorter time. For this reason the objective of the present study was to optimize the procedure to determine the sensitivity and specificity limits for the detection of L. monocytogenes from fresh cheese and the predictive value of the test. In order to achieve this objective, 76 pasteurized cheese samples were evaluated (45 samples were artificially inoculated at the lab and 31 were used as negative controls). The validation of the technique was done in 50 samples of non pasteurized fresh cheese. Traditional culture isolation was performed according to the methodology described in Compendium of Methods for the Microbiological Examination of Foods. PCR reaction for the detection of L. monocytogenes was based on the methodology described by Poutou,using primers characteristic of the genus and the listeriolisine O gene that is specie's specific. The optimal incubation period determined for the selective enrichment broth was of 48h, and a 100% sensitivity, specificity, predictive value (positive and negative) were obtained by PCR. The technique validation showed the specificity ofthe test in the detection of only the L. monocytogenes species, and not other genus or species that may appear in food matrixes or in food environments.

[Microbiological evaluation of ready-to-eat foods manufactured by small Costa Rican industries]. / Evaluación microbiológica de alimentos listos para consumo procesados por pequeñas industrias costarricenses.

Ready-to-eat (RTE) foods are processed foodstuffs which have gained popularity in recent times because they can be ingested without further thermic treatments. In this work, the microbiological quality and safety of 90 samples of RTE foods manufactured by small Costa Rican industries was determined to evaluate whether they represent a Public Health risk. Twenty-six samples of pickled vegetables, 18 dips, 18 salads, and 12 sweet treats were studied. Each sample was analyzed with regard to its pH, the presence of culturable microbiological quality indicators and recognized foodborne pathogens (Salmonella, Listeria monocytogenes. Clostridium perfringens, C. botulinum, and Bacillus cereus) Selected genes encoding toxins of C. botulinum and C. perfringens were screened by PCR. Thirty-seven percent of the samples had a level of acidity that could allow the growth and proliferation of bacterial pathogens (pH > or = 4.5). The shelf-life indicators were acceptable but only if the RTE foods are kept at adequate conditions of temperature and humidity. Sixty-four percent of the RTE foods had total coliforms values that evidence inadequate hygiene practices during its elaboration (MPN/g >1000). This result was confirmed by the finding of fecal coliforms in 56% of the samples, which, by the way, are inacceptable for human consumption. All cultures for pathogens were negative, except for 4 samples that contained B. cereus. Toxins of C. botulinum were not detected and one single sample was positive for the PCR for C. perfringens. The elevated degree of fecal contamination detected in the RTE could be prevented by means of good manufacturing practices, better hygiene measures and a deeper attention to critical control points.

Crystal structures and biological activity of 1,1,4-triphenyl-substituted 1,3-enyne compounds.

1,3-Enyne structural motifs are versatile building blocks in organic synthesis and occur widely in various natural products with many of them being highly active as cytotoxic macrolides and antitumour antibiotics. This article presents the crystal structure of three 1,1,4-triphenyl-substituted 1,3-enynes, viz. 4-(2-methylphenyl)-1,1-diphenylbut-1-en-3-yne, C23H18 (1), 4-(2-methoxyphenyl)-1,1-diphenylbut-1-en-3-yne, C23H18O (2), and 4-(4-nitrophenyl)-1,1-diphenylbut-1-en-3-yne, C22H15NO2 (3). The benzene ring at position 4 of the but-1-en-3-yne group bears a weakly activating methyl group in compound 1, a moderately activating methoxy group in 2 and a strongly deactivating nitro group in 3. The crystal structures of 1 and 3 both have monoclinic symmetry, while that of 2 is orthorhombic, and all of them have one molecule in the asymmetric unit. All three compounds were investigated for their antibacterial and antifungal activities. Interestingly, enyne 2 is the only compound tested that inhibited the growth of Aspergillus niger.

[Characterization of Listeria monocytogenes isolates obtained from raw cheese samples acquired from different Costa Rican producer zones]. / Caracterización de cepas de Listeria monocytogenes realizados a partir de queso fresco proveniente de diferentes zonas productoras costarricenses.

In Costa Rica, almost 25% of the national milk production is used for the elaboration of non pasteurized soft cheese, and the annual intake of this product is around 4-5 kg per capita. This product has been identified as the source of food borne outbreaks due to Listeria monocytogenes. Given that, the isolation and identification of this bacterium from non pasteurized soft cheese samples coming from two producer zones of Costa Rica was performed. 110 cheese samples were collected, from which 27 L. monocytogenes strains were isolated. These were characterized using biochemical and serological tests, also, susceptibility to common used antibiotics, test tube hemolysis and invasion in Hela cells trials were performed. 85% of the strains evaluated were sensible to all the antibiotics analyzed, nevertheless, four strains presented resistance to different agents, including streptomycin, kanamycin, cephalotin and tetracycline. Also, multiple resistance patterns were found. 88.9% of the studied isolates were positive for the test tube hemolysis trial; 22.2% presented invasion percentages higher than the clinical origin strain used as control. It is important to point out that all the invasive strains were completely susceptible to the antibiotics tested. The results found demonstrate the presence of L. monocytogenes in Costa Rican soft cheese samples. Also, demonstrate its high percent of susceptibility to common use antibiotics. Same time, invasion trials show that soft cheese may be a source of invasive and potentially pathogenic strains for human being.

[Identification, quantification and antimicrobial susceptibility pattern of probiotic bacteria added to common use food products in Costa Rica]. / Identificación, cuantificación y determinación del perfil de sensibilidad a antibióticos de bacterias prebióticas adicionadas a productos de consumo frecuente en Costa Rica.

In the last years, due to the high demand of food products supplemented with probiotics and the multiple nutritional and therapeutic benefits associated with them, research on these microorganisms has advanced considerably, including their selection and characterization. As a general recommendation, several entities as World Health Organization (WHO) and United Nations Organization for Agriculture and Food recommend that the specification of the alive species contained and their number shall appear in the label of the product. In the present study, six different commercially available products, supplemented with probiotics were analyzed, in order to evaluate the concentration of microorganisms through the shelf life of the product, identify the strains isolated and determine the antibiotic susceptibility pattern of these. Results demonstrated that the strains isolated kept acceptable concentrations during the 28 days of storage. Nevertheless, the identification of these strains variated from the one reported on the label on several of the products tested. This can be due to the commercial method used for the identifications, which is based in the carbohydrate fermentation pattern and not in genotypic trials. The antimicrobials' susceptibility patterns found show that further research shall be performed in order to establish the intrinsic or acquired nature of the resistance determinants, and if these are codified by transferable elements among bacteria.

[Toxigenic Bacillus cereus detection in lactic products with spices and dehydrated milk collected in Costa Rica]. / Detección de Bacillus cereus toxigénicos en productos láacteos con especias y leches deshidratadas colectadas en Costa Rica.

Bacillus cereus is a Gram positive rod widely distributed in nature and associated to different types of food that, under some circumstances, may cause pathology to human beings. Diarrheic and emetic strains have been described based on the type of toxins produced. In order to determine the risk to health represented by this bacteria, the toxigenic potential of strains isolated from cheese with spices, spread cheese with spices and dehydrated milk, all sold in San José, Costa Rica, were determined using a multiplex PCR technique with oligonucleotides specific for the genes coding toxins HBL and Nhe. From 45 samples collected, 15 isolates of B cereus were obtained (60% coming from spread cheese with spices 7% from dehydrated milk and 13% from cheese with spices). All the strains analyzed presented at least one of the genes analyzed; six of them, coming from dehydrated milk and spread cheese, showed molecular evidence of the genes nheB, nheA, nheC, hblD, hblA y hblC, confirming the correlation described for the presence of operons codifying for HBL and Nhe. Nevertheless, the no detection of a gene cannot be considered as a definitive proof of its absence, given the existence of polymorphism in the sequences of the genes analyzed. The results obtained show that multiple of the B cereus strains found in lactic products from Costa Rica have the necessary genes for synthesizing toxins, so the correct handling of these products is very important since they can represent a risk for public health.

[Bacteriological evaluation of goat milk and cheese distributed in the Metropolitan Area of San José, Costa Rica]. / Evaluación bacteriológica de la leche y queso de cabra distribuidos en el Area Metropolitana de San José, Costa Rica.

In the last years, there has been an increase in the production, industrialization and consumption of goat's milk and derivate products, including cheese, worldwide. Nevertheless, in Costa Rica there is no study of these products, reason why the objective of this work was to determine the microbiological characteristics of goat's milk and fresh cheese distributed in the Metropolitan Area of San José, Costa Rica, in order to evaluate its impact in the economical field and as a potential risk for Public Health. A total of 25 raw goat's milk samples, obtained by manual milking from 5 different producers tested in five different dates and 15 cheese samples, elaborated with pasteurized milk, commercially available and coming from three different producers were analyzed. The study included the analysis of spoilage bacteria (total aeobic count and lactic bacteria count), indicators of hygiene (total coliforms), fecal contamination (fecal coliforms), manipulation (Staphylococcus aureus) and pathogens (Listeria monocytogenes and Salmonella spp). High results were obtained for the total aerobic count and lactic bacteria count of the milk and cheese samples, showing a reduced shelf life. Total coliforms, in limits beyond the established ones by the Costa Rican legislation for human consumption raw milk, were found in 100% of milk samples, as well as for fecal coliforms in 76% of them. All cheese samples, except one, were negative for these indicators, suggesting good manufacturing practices. S. aureus counts were low and both Salmonella spp. and L. monocytogenes were not isolated from samples analyzed.

[Staphylococcus aureus enterotoxin A detection using the polymerase chain reaction (PCR) and its correlation with coagulase and thermonuclease tests]. / Deteccion de la enterotoxina A de Staphylococcus aureus mediante la reaccion en cadena de la polimerasa (PCR) y su correlacion con las pruebas de coagulasa y termonucleasa.

Staphylococcus aureus is a pathogenic bacterium, widely distributed on nature and associated to general infection and food borne outbreaks. The relationship between this bacterium and food borne outbreaks has been done, historically, using several tests, including coagulase, thermonuclease and actually, PCR for the genes codifying for the enterotoxin responsible of clinical symptoms. The objective of this work is to detect enterotoxin A codifying gene through PCR in a group of S. aureus strains isolated from food samples, and also to correlate the presence of this gene with the production of coagulase and thermonuclease enzymes. A total of 69 staphylococcal strains were analyzed, 58 obtained from non pasteurized milk samples from the Estación Experimental Alfredo Volio Mata and 11 from the Food and Water Microbiology Laboratory collection, Universidad de Costa Rica. Coagulase, thermonuclease and enterotoxin A were analyzed in all the strains, and a statistical correlation was performed in order to verify possible associations. Results show that there is no correlation between the three variables, nevertheless, all coagulase positive strains were thermonuclease positive, and all enterotoxin positive strains were coagulase and thermonuclease positive, but not inversely. These results show that the use of presumptive or indirect tests for establishing entorotoxigenity of S. aureus strains is not truthful, more sensible and specific analysis, as PCR, shall be performed.

[Microbiological contamination and antimicrobial activity of cristalised cane sugar on some medically important microorganisms in Costa Rica]. / Contaminación microbiológica y actividad antimicrobiana del azúcar de caña cristalizado sobre algunos agentes de interés médico en Costa Rica.

Microbiological contamination and antimicrobial activity of cristalised cane sugar on some medically important microorganisms in Costa Rica. Unrefined cristalised cane sugar, obtained after the filtration and evaporation of sugar cane juice, is a nutritional product of traditional consumption in Costa Rica and other Neotropical countries. It has been used in the topic treatment of infected wounds, with satisfactory results even with some antibiotic-ressistant bacteria. We studied the microbiological quality of 50 commercial samples. The analyses included total aerobic and anaerobic bacteria plate count; aerobic and anaerobic spore count; mold and yeast count; total and fecal coliforms; and presence of Clostridium botulinum. The antimicrobial effect was tested for Staphylococcus aureus (ATCC 25923), S. epidermidis (UCR 2902), Pseudomonas aeruginosa (ATCC 9027), Escherichia coli (ATCC 25922), Salmonella enteritidis (ATCC 13076), Listeria monocytogenes (ATCC 19116) and Aspergillus niger (Asni 06). Most of the samples (76%) presented counts lower than 100 CFU/g especially for sporulated forms (90% lower than 20 CFU/g), the mold and yeast count was higher (38% higher than 10(2) CFU/g), demonstrating the importance of these microorganisms in the spoilage of the product; 76% of the samples presented fecal contamination; C. botulinum was not isolated with the methodology employed. No inhibitory effect was observed for A. niger, but all samples han an inhibitory effect over the other species, especially for P. aeruginosa and S. aureus.

[Evaluation of the effect of Lactobacillus rhamnosus probiotic culture added to yogurt over Staphylococcus aureus, Escherichia coli O157:H7, Listeria monocytogenes and Salmonella enteritidis populations]. / Evaluación del efecto del cultivo probiótico Lactobacillus rhamnosus adicionado a yogurt natural y con probi6ticos comerciales sobre poblaciones de Staphylococcus aureus, Escherichia coli O157:H7, Listeria monocytogenes y Salmonella enteritidis.

The effect of different types of probiotics present in yogurt over known populations of Staphylococcus aureus, Escherichia coli O157:H7, Listeria monocytogenes and Salmonella enteritidis was evaluated. The three types of yogurt used were: without added probiotics, with added probiotics (Lactobacillus casei CRL_431 and L. acidophilus CRL_730 CHR HANSEN) and another one with the same probiotics mentioned above and Lactobacillus rhamnosus (LR-35) culture. About 10(9) CFU/ mL of each potentially pathogenic bacteria was added to each type of yogurt tested, and kept in refrigeration at 4 degrees C during its shelf life, about 30 days. Bacterial count was done the initial day and every four days. Results obtained show that there is a difference in the inhibition between yogurts without added probiotics and the commercial yogurt with added probiotics; there is a clear inhibitory effect of the last one over S. aureus, E. coli O157:H7 and Listeria monocytogenes. The yogurt with added probiotics and L. rhamnosus did not show any additional inhibitory effect over the bacteria tested when compared with the yogurt with added probiotics. S. enteritidis could not be evaluated because it was not detectable in any yogurt samples evaluated four days after its inoculation. This study confirms the antagonic effect of probiotic cultures over potentially pathogenic bacteria for human beings and animals that may be present in food. Nevertheless, the use of L. rhamnosus did not produce any additional inhibitory effect.

Isolation and Identification of Arcobacter Species from Costa Rican Poultry Production and Retail Sources.

Arcobacter is a gram-negative rod recognized as a potential food- and waterborne pathogen; nevertheless, little is known about the effects of this pathogen on human and animal health. Although Arcobacter species are commonly found in nature, poultry is suspected to be the main vehicle for the transmission of this pathogen. The aims of this work were to determine the prevalence of Arcobacter spp. in broilers produced in Costa Rica for human consumption and to analyze the pathogenic capacity of the isolates through the detection of virulence genes. One hundred fifty-two samples of cecal content (87 farms), 104 samples of carcass rinse after chiller (six processing plants), and 96 carcass rinses from as many retail stores were analyzed. The suspicious isolates were identified using genus-specific PCR, and species-level identification was achieved with a multiplex PCR. Virulence genes were identified using the protocol described by L. Douidah, L. de Zutter, J. Baré, P. De Vos, P. Vandamme, O. Vandenberg, A.-M. Van den Abeele, and K. Houf (J. Clin. Microbiol. 50:735-741, 2012), which includes nine different virulence genes. The overall isolation frequency of Arcobacter was 6.5% (n = 23). Eight (34.8%) of the isolates came from cecal content, 2 (8.7%) were isolated from samples taken after chiller, and 13 (56.5%) were from retail stores. The species isolated included A. thereius (30.4%), A. butzleri (21.7%), A. skirrowii (4.3%), and A. cibarius (4.3%). The remaining samples were classified as Arcobacter sp. Gene tlyA was the most prevalent virulence gene, present in 9 of 23 samples analyzed; genes hecA and pldA were present in one only strain each. A strain of A. butzleri isolated from a retail store presented the highest number of virulence genes (five), and 11 samples did not present any of the genes analyzed. The results obtained suggest that the presence of virulent Arcobacter isolates in the poultry production chain from Costa Rica could be a risk for individuals who consume the contaminated product.

[Comparison of the antibiotics sensibility pattern of Listeria monocytogenes and Salmonella spp. strains isolated from food with clinical origin samples]. / Comparación del perfil de sensibilidad a antibióticos de cepas de Listeria monocytogenes y Salmonella spp. aisladas a partir de alimentos con cepas de origen clínico.

The actual use of antibiotics includes, not just its therapeutic cases, but also for disease prevention and as a growth promoter in animals. These practices have resulted in the propagation of resistance to antibiotics, representing a threat for Public Health. In this work, the antibiotic sensibility pattern of 20 Listeria monocytogenes and 40 Salmonella spp. strains, isolated from foodstuff was studied and compared with the antibiotic sensibility patterns of 20 L. monocytogenes and 100 Salmonella strains of clinical origin. 95% of the L. monocytogenes strains isolated from food were sensible to ampicillin, compared with the 65% of the clinical origin strains. Same way, 100% of food strains were sensible to gentamicin, compared with 85% of clinical origin strains. 95% of both showed sensibility to trimethoprim sulfametoxazole and 100% to ciprofloxacin. For Salmonella spp., the sensibility patterns for trimethoprim sulfametoxazole, gentamicin, ciprofloxacin, nalidixic acid and amoxicilin/clavulanic acid from both origins were similar. Nevertheless, food origin strains showed a 97.5% and 82.5% sensibility for tetracycline and cephalosporin respectively, compared with a 83 and 90% sensibility shown by clinical origin strains. The results obtained demonstrate the potential risk that bacterial strains isolated from food represent in the transmission of antibiotics' resistance.

Genes that encode botulism neurotoxins A, B, E and F in neotropical bee honey identified with the polymerase chain reaction.

Honey can be used for the treatment of wounds, sores and skin bums, but it might be contaminated with Clostridium botulinum spores. In order to evaluate Costa Rican raw honey samples, the detection of neurotoxin gene sequences (corresponding to the bacterium) C. botulinum A, B, E and F was done with the polymerase chain reaction. A total of 64 raw honey samples, coming from different Costa Rican sites were analyzed. Reference C. botulinum strains type A (ATCC 19397), type B (ATCC 7949), type E (ATCC 17786) and type F (ATCC 25764) were used as templates for testing the effectivity of the method. The process consisted in culturing the honey samples in prereduced triptose-peptone-glucose-yeast extract media (TGPY) for 5 days. After this, the bacteria lysate obtained was used for PCR. The amplicons, product of the reaction, were visualized using agarose gel 2%. From the 64 honey samples analyzed, none produced positive results in the PCR, since no amplicons were obtained. Even though, all the reference C. botulinum strains used as controls were visualized and showed the effectivity of the extraction method and of the PCR used. The results obtained show promising therapeutic uses for honey from Costa Rica, but further evaluations shall be done in order to be sure of the safety of the product.