Listeria monocytogenes is causing listeriosis, a rare but severe foodborne infection. Listeriosis... more Listeria monocytogenes is causing listeriosis, a rare but severe foodborne infection. Listeriosis affects pregnant women, newborns, older adults, and immunocompromised individuals. Ready-to-eat (RTE) foods are the most common sources of transmission of the pathogen This study explored the virulence factors and antibiotic resistance in L. monocytogenes strains isolated from ready-to-eat (RTE) foods through in vitro and in silico testing by whole-genome sequencing (WGS). The overall positivity of L. monocytogenes in RTE food samples was 3.1% and 14 strains were isolated. L. monocytogenes ST8, ST2763, ST1, ST3, ST5, ST7, ST9, ST14, ST193, and ST451 sequence types were identified by average nucleotide identity, ribosomal multilocus sequence typing (rMLST), and core genome MLST. Seven isolates had serotype 1/2a, five 1/2b, one 4b, and one 1/2c. Three strains exhibited in vitro resistance to ampicillin and 100% of the strains carried the fosX, lin, norB, mprF, tetA, and tetC resistance ge...
Background: One of the most common pathogens causing alimentary toxi-infections is Staphylococcus... more Background: One of the most common pathogens causing alimentary toxi-infections is Staphylococcus aureus (S aureus). The presence of S aureus in food, indicates flaws during food manipulations. For this reason most sanitary norms require the detection of S aureus carriers. Aim: To determine the carriage rate of enterotoxin producing S aureus strains in food handlers, and to evaluate the antibiotic susceptibility to six antimicrobial agents. Materials and Methods: A total of 102 food handlers from 19 restaurants in Santiago, were analyzed. Samples for microbiological analysis were obtained with a swab from the retropharynx. Results: S aureus grew in 35 out of the 102 samples obtained (34%). Further analysis revealed that 19/35 (54%) strains were able to produce enterotoxins. Therefore the corrected carriage rate was 19% (19/102). The most frequently detected enterotoxin was the type A (12/19). All S aureus isolates were resistant to penicillin and susceptible to oxacillin, clindamyci...
Cronobacter sakazakii is an enteropathogen that causes neonatal meningitis, septicemia, and necro... more Cronobacter sakazakii is an enteropathogen that causes neonatal meningitis, septicemia, and necrotizing enterocolitis in preterm infants and newborns with a mortality rate of 15 to 80%. Powdered and dairy formulas (P-DF) have been implicated as major transmission vehicles and subsequently the presence of this pathogen in P-DF led to product recalls in Chile in 2017. The objective of this study was to use whole genome sequencing (WGS) and laboratory studies to characterize Cronobacter strains from the contaminated products. Seven strains were identified as C. sakazakii, and the remaining strain was Franconibacter helveticus. All C. sakazakii strains adhered to a neuroblastoma cell line, and 31 virulence genes were predicted by WGS. The antibiograms varied between strains. and included mcr-9.1 and blaCSA genes, conferring resistance to colistin and cephalothin, respectively. The C. sakazakii strains encoded I-E and I-F CRISPR-Cas systems, and carried IncFII(pECLA), Col440I, and Col(pH...
In this study we evaluated if zebrafish larvae can be colonized by human gut microorganisms. We t... more In this study we evaluated if zebrafish larvae can be colonized by human gut microorganisms. We tested two strategies: (1) through transplantation of a human fecal microbiota and (2) by successively transplanting aerotolerant anaerobic microorganisms, similar to the colonization in the human intestine during early life. We used conventionally raised zebrafish larvae harboring their own aerobic microbiota to improve the colonization of anaerobic microorganisms. The results showed with the fecal transplant, that some members of the human gut microbiota were transferred to larvae. , one unclassified genus of the family Ruminococcaceae and Enterobacteriaceae were detected in 3 days post fertilization (dpf) larvae; however only persisted to 7 dpf. Successive inoculation of and did not improve their colonization, compared to individual inoculation of each bacterial species. Interestingly, the sporulating bacteria and were the most persistent microorganisms. Their endospores persisted at l...
The aims of this study were to determine the occurrence of Listeria monocytogenes in cattle feces... more The aims of this study were to determine the occurrence of Listeria monocytogenes in cattle feces and ground beef, to characterize these strains by pulsed-field gel electrophoresis and to compare them to three listeria strains found in humans. Cattle from different origins (n = 250) and ground beef obtained from supermarkets (n = 40) were sampled. The results show low occurrence in cattle feces (0.4 %) but a higher presence in ground beef (37 %). An important part of the ground beef strains (80 %) had > 95 % similarity with a strain isolated from a human sporadic case and the ATCC 19115 used as control. The strain isolated from cattle feces had 93 % similarity to clone 009, previously associated with a listeriosis outbreak related to cheese. Cattle and ground beef can harbor virulent L. monocytogenes strains. Further studies in animals and animal products are needed to improve listeriosis control.
Measurement of changes in serum antibodies is an excellent predictor of Helicobacter pylori eradi... more Measurement of changes in serum antibodies is an excellent predictor of Helicobacter pylori eradication after antibiotic treatment. To measure the changes in serum antibody titers to Helicobacter pylori, before and after treatment. IgG antibodies to H. pylori were prospectively evaluated in 107 duodenal ulcer patients treated either with antibiotics (amoxicillin, metronidazole and bismuth subsalicylate) plus omeprazole or omeprazole alone. IgG antibody levels were determined using an "in house" ELISA in sera from 49 eradicated patients that received quadruple therapy and 58 non-eradicated patients (12 in whom antibiotic therapy failed and 46 that received omeprazole alone). Endoscopy, urease test, microscopy, and culture of gastric biopsies confirmed H. pylori eradication. Patients in whom H. pylori was eradicated, showed a maintained drop in serum antibody titers that ranged from 15%, 62%, 74% to 76% at 28 days, 4, 8 and 12 months respectively. Such reduction was not obse...
The efficiency of an ELISA method, designed to detect polyvalent IgG and IgM antibodies to Salmon... more The efficiency of an ELISA method, designed to detect polyvalent IgG and IgM antibodies to Salmonella typhi polysaccharide was evaluated in patients admitted or convalescing from typhoid fever and in control subjects. Polyvalent antibodies to S typhi were demonstrated in 28/30 (93%) typhoid patients, 0/11 bacteremic patients (E coli or S paratyphi A) and 0/15 asymptomatic individuals. Widal test showed significant anti-0 agglutinin values (> = 1: 160) in only 12/30 (40%), 1/11 (9%) and 0/15 subjects from each group respectively. Typhoid patients tested on admission or at discharge showed similar high reactivity rates to ELISA. On the contrary, the Widal test detected only 2/15 (13%) patients on admission (p < 0.02) and 10/15 (67%) at discharge. These results and additional immunoblotting tests suggest that ELISA developed to detect polyvalent anti-LPS antibodies could represent a highly specific diagnostic tool to confirm typhoid fever in endemic areas.
The objective of this study was to evaluate the prevalence of antibodies to Helicobacter pylori C... more The objective of this study was to evaluate the prevalence of antibodies to Helicobacter pylori CagA and VacA proteins and correlate this prevalence with gastric diseases in colonised Chileans. The study was performed in 418 adults colonised with H. pylori: 316 with gastroduodenal pathology (152 duodenal ulcer, 14 gastric cancer and 150 gastritis patients) and 102 asymptomatic subjects. Serum IgG antibodies to H. pylori were determined by enzyme immunoassay (EIA). Antibodies to VacA and CagA proteins were detected by Western blotting. In a subgroup of the patients, the vacuolating activity was determined by HeLa cell assay and the CagA product was confirmed by PCR assay. IgG antibodies to both VacA and CagA proteins of H. pylori were found in 270 (85%) of 316 colonised gastric patients and in 72 (71%) of 102 asymptomatic subjects. Colonisation with virulent strains was significantly higher among duodenal ulcer and gastric cancer patients than in gastritis patients or asymptomatic su...
ABSTRACT The influence of chronic iron intake on the incidence and aetiology of acute diarrhoea w... more ABSTRACT The influence of chronic iron intake on the incidence and aetiology of acute diarrhoea was prospectively evaluated in a group of Chilean infants. A total of 430 spontaneously weaned infants, aged 3–12 mths, were fed either iron-fortified (IF) or non-enriched (NE) milk formulae. The 6 mths period of continuous surveillance was accomplished through biweekly home visits by trained nurses. Diarrhoeal episodes were recorded and stool samples obtained from symptomatic and paired asymptomatic infants. Results showed a similar incidence of diarrhoeal disease among infants fed IF and NE formulae (1·38 and 1·27 cases/child/6 mths respectively). The isolation rate and pathogen distribution were also similar in both groups, either during diarrhoeal episodes (44 per cent and 45 per cent) or asymptomatic infection (19 per cent and 16 per cent). In addition, the number of diarrhoeal episodes associated with mixed bacterial pathogens or with protracted excretion (&gt; 7 d) was apparently not influenced by the ingestion of iron-supplemented formula. These results suggest that chronic consumption of iron-enriched formula neither enhances the incidence of diarrhoea nor modifies the intestinal bacterial ecology in infants who live in areas with suboptimal sanitary conditions.
To determine the distributions of six Helicobacter pylori (Hp)-specific antibodies in a high-risk... more To determine the distributions of six Helicobacter pylori (Hp)-specific antibodies in a high-risk population of gastric cancer (GC) and explore the relationship between Hp virulence factors and precancerous gastric lesions. Based on the two intervention trials conducted in Linqu County, the seropositivities for CagA, VacA, GroEL, UreA, HcpC and GGT were assessed by recombinant immunoassay (recomLine) in 623 participants with H. pylori infection determined by (13)C-urea breath test ((13)C-UBT) and/or enzyme linked immunosorbent assay (ELISA). In a total of 623 participants were detected by recomLine analysis, of which 594 were Hp-positive. The seropositivities rates of CagA, VacA, GroEL, UreA, HcpC and GGT were 84.0%, 38.2%, 66.7%, 17.7%, 58.8% and 42.8%, respectively. A total of 523 participants were determined as type I infection of Hp, accounting for 88.1%. Compared with superficial gastritis (SG), the infection rate of Hp type I was higher in the chronic atrophic gastritis (CAG) (P = 0.001). The results of this population-based study suggest that the virulence factors of Hp may be related to the development of GC in a Chinese high-risk population. The recomLine analysis may serve as a tool for identification of Hp strains and prediction of high-risk population of GC.
Listeria monocytogenes is causing listeriosis, a rare but severe foodborne infection. Listeriosis... more Listeria monocytogenes is causing listeriosis, a rare but severe foodborne infection. Listeriosis affects pregnant women, newborns, older adults, and immunocompromised individuals. Ready-to-eat (RTE) foods are the most common sources of transmission of the pathogen This study explored the virulence factors and antibiotic resistance in L. monocytogenes strains isolated from ready-to-eat (RTE) foods through in vitro and in silico testing by whole-genome sequencing (WGS). The overall positivity of L. monocytogenes in RTE food samples was 3.1% and 14 strains were isolated. L. monocytogenes ST8, ST2763, ST1, ST3, ST5, ST7, ST9, ST14, ST193, and ST451 sequence types were identified by average nucleotide identity, ribosomal multilocus sequence typing (rMLST), and core genome MLST. Seven isolates had serotype 1/2a, five 1/2b, one 4b, and one 1/2c. Three strains exhibited in vitro resistance to ampicillin and 100% of the strains carried the fosX, lin, norB, mprF, tetA, and tetC resistance ge...
Background: One of the most common pathogens causing alimentary toxi-infections is Staphylococcus... more Background: One of the most common pathogens causing alimentary toxi-infections is Staphylococcus aureus (S aureus). The presence of S aureus in food, indicates flaws during food manipulations. For this reason most sanitary norms require the detection of S aureus carriers. Aim: To determine the carriage rate of enterotoxin producing S aureus strains in food handlers, and to evaluate the antibiotic susceptibility to six antimicrobial agents. Materials and Methods: A total of 102 food handlers from 19 restaurants in Santiago, were analyzed. Samples for microbiological analysis were obtained with a swab from the retropharynx. Results: S aureus grew in 35 out of the 102 samples obtained (34%). Further analysis revealed that 19/35 (54%) strains were able to produce enterotoxins. Therefore the corrected carriage rate was 19% (19/102). The most frequently detected enterotoxin was the type A (12/19). All S aureus isolates were resistant to penicillin and susceptible to oxacillin, clindamyci...
Cronobacter sakazakii is an enteropathogen that causes neonatal meningitis, septicemia, and necro... more Cronobacter sakazakii is an enteropathogen that causes neonatal meningitis, septicemia, and necrotizing enterocolitis in preterm infants and newborns with a mortality rate of 15 to 80%. Powdered and dairy formulas (P-DF) have been implicated as major transmission vehicles and subsequently the presence of this pathogen in P-DF led to product recalls in Chile in 2017. The objective of this study was to use whole genome sequencing (WGS) and laboratory studies to characterize Cronobacter strains from the contaminated products. Seven strains were identified as C. sakazakii, and the remaining strain was Franconibacter helveticus. All C. sakazakii strains adhered to a neuroblastoma cell line, and 31 virulence genes were predicted by WGS. The antibiograms varied between strains. and included mcr-9.1 and blaCSA genes, conferring resistance to colistin and cephalothin, respectively. The C. sakazakii strains encoded I-E and I-F CRISPR-Cas systems, and carried IncFII(pECLA), Col440I, and Col(pH...
In this study we evaluated if zebrafish larvae can be colonized by human gut microorganisms. We t... more In this study we evaluated if zebrafish larvae can be colonized by human gut microorganisms. We tested two strategies: (1) through transplantation of a human fecal microbiota and (2) by successively transplanting aerotolerant anaerobic microorganisms, similar to the colonization in the human intestine during early life. We used conventionally raised zebrafish larvae harboring their own aerobic microbiota to improve the colonization of anaerobic microorganisms. The results showed with the fecal transplant, that some members of the human gut microbiota were transferred to larvae. , one unclassified genus of the family Ruminococcaceae and Enterobacteriaceae were detected in 3 days post fertilization (dpf) larvae; however only persisted to 7 dpf. Successive inoculation of and did not improve their colonization, compared to individual inoculation of each bacterial species. Interestingly, the sporulating bacteria and were the most persistent microorganisms. Their endospores persisted at l...
The aims of this study were to determine the occurrence of Listeria monocytogenes in cattle feces... more The aims of this study were to determine the occurrence of Listeria monocytogenes in cattle feces and ground beef, to characterize these strains by pulsed-field gel electrophoresis and to compare them to three listeria strains found in humans. Cattle from different origins (n = 250) and ground beef obtained from supermarkets (n = 40) were sampled. The results show low occurrence in cattle feces (0.4 %) but a higher presence in ground beef (37 %). An important part of the ground beef strains (80 %) had > 95 % similarity with a strain isolated from a human sporadic case and the ATCC 19115 used as control. The strain isolated from cattle feces had 93 % similarity to clone 009, previously associated with a listeriosis outbreak related to cheese. Cattle and ground beef can harbor virulent L. monocytogenes strains. Further studies in animals and animal products are needed to improve listeriosis control.
Measurement of changes in serum antibodies is an excellent predictor of Helicobacter pylori eradi... more Measurement of changes in serum antibodies is an excellent predictor of Helicobacter pylori eradication after antibiotic treatment. To measure the changes in serum antibody titers to Helicobacter pylori, before and after treatment. IgG antibodies to H. pylori were prospectively evaluated in 107 duodenal ulcer patients treated either with antibiotics (amoxicillin, metronidazole and bismuth subsalicylate) plus omeprazole or omeprazole alone. IgG antibody levels were determined using an "in house" ELISA in sera from 49 eradicated patients that received quadruple therapy and 58 non-eradicated patients (12 in whom antibiotic therapy failed and 46 that received omeprazole alone). Endoscopy, urease test, microscopy, and culture of gastric biopsies confirmed H. pylori eradication. Patients in whom H. pylori was eradicated, showed a maintained drop in serum antibody titers that ranged from 15%, 62%, 74% to 76% at 28 days, 4, 8 and 12 months respectively. Such reduction was not obse...
The efficiency of an ELISA method, designed to detect polyvalent IgG and IgM antibodies to Salmon... more The efficiency of an ELISA method, designed to detect polyvalent IgG and IgM antibodies to Salmonella typhi polysaccharide was evaluated in patients admitted or convalescing from typhoid fever and in control subjects. Polyvalent antibodies to S typhi were demonstrated in 28/30 (93%) typhoid patients, 0/11 bacteremic patients (E coli or S paratyphi A) and 0/15 asymptomatic individuals. Widal test showed significant anti-0 agglutinin values (> = 1: 160) in only 12/30 (40%), 1/11 (9%) and 0/15 subjects from each group respectively. Typhoid patients tested on admission or at discharge showed similar high reactivity rates to ELISA. On the contrary, the Widal test detected only 2/15 (13%) patients on admission (p < 0.02) and 10/15 (67%) at discharge. These results and additional immunoblotting tests suggest that ELISA developed to detect polyvalent anti-LPS antibodies could represent a highly specific diagnostic tool to confirm typhoid fever in endemic areas.
The objective of this study was to evaluate the prevalence of antibodies to Helicobacter pylori C... more The objective of this study was to evaluate the prevalence of antibodies to Helicobacter pylori CagA and VacA proteins and correlate this prevalence with gastric diseases in colonised Chileans. The study was performed in 418 adults colonised with H. pylori: 316 with gastroduodenal pathology (152 duodenal ulcer, 14 gastric cancer and 150 gastritis patients) and 102 asymptomatic subjects. Serum IgG antibodies to H. pylori were determined by enzyme immunoassay (EIA). Antibodies to VacA and CagA proteins were detected by Western blotting. In a subgroup of the patients, the vacuolating activity was determined by HeLa cell assay and the CagA product was confirmed by PCR assay. IgG antibodies to both VacA and CagA proteins of H. pylori were found in 270 (85%) of 316 colonised gastric patients and in 72 (71%) of 102 asymptomatic subjects. Colonisation with virulent strains was significantly higher among duodenal ulcer and gastric cancer patients than in gastritis patients or asymptomatic su...
ABSTRACT The influence of chronic iron intake on the incidence and aetiology of acute diarrhoea w... more ABSTRACT The influence of chronic iron intake on the incidence and aetiology of acute diarrhoea was prospectively evaluated in a group of Chilean infants. A total of 430 spontaneously weaned infants, aged 3–12 mths, were fed either iron-fortified (IF) or non-enriched (NE) milk formulae. The 6 mths period of continuous surveillance was accomplished through biweekly home visits by trained nurses. Diarrhoeal episodes were recorded and stool samples obtained from symptomatic and paired asymptomatic infants. Results showed a similar incidence of diarrhoeal disease among infants fed IF and NE formulae (1·38 and 1·27 cases/child/6 mths respectively). The isolation rate and pathogen distribution were also similar in both groups, either during diarrhoeal episodes (44 per cent and 45 per cent) or asymptomatic infection (19 per cent and 16 per cent). In addition, the number of diarrhoeal episodes associated with mixed bacterial pathogens or with protracted excretion (&gt; 7 d) was apparently not influenced by the ingestion of iron-supplemented formula. These results suggest that chronic consumption of iron-enriched formula neither enhances the incidence of diarrhoea nor modifies the intestinal bacterial ecology in infants who live in areas with suboptimal sanitary conditions.
To determine the distributions of six Helicobacter pylori (Hp)-specific antibodies in a high-risk... more To determine the distributions of six Helicobacter pylori (Hp)-specific antibodies in a high-risk population of gastric cancer (GC) and explore the relationship between Hp virulence factors and precancerous gastric lesions. Based on the two intervention trials conducted in Linqu County, the seropositivities for CagA, VacA, GroEL, UreA, HcpC and GGT were assessed by recombinant immunoassay (recomLine) in 623 participants with H. pylori infection determined by (13)C-urea breath test ((13)C-UBT) and/or enzyme linked immunosorbent assay (ELISA). In a total of 623 participants were detected by recomLine analysis, of which 594 were Hp-positive. The seropositivities rates of CagA, VacA, GroEL, UreA, HcpC and GGT were 84.0%, 38.2%, 66.7%, 17.7%, 58.8% and 42.8%, respectively. A total of 523 participants were determined as type I infection of Hp, accounting for 88.1%. Compared with superficial gastritis (SG), the infection rate of Hp type I was higher in the chronic atrophic gastritis (CAG) (P = 0.001). The results of this population-based study suggest that the virulence factors of Hp may be related to the development of GC in a Chinese high-risk population. The recomLine analysis may serve as a tool for identification of Hp strains and prediction of high-risk population of GC.
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