Common presence of Helicobacter DNA in the gallbladder of patients with gallstone diseases and controls

BACKGROUND: Several species of Helicobacter colonise the biliary tract of animals and cause hepatobiliary diseases. Helicobacter species have also been identified in the gallbladder of a high proportion of Chilean patients with gallbladder cancer. AIM: To determine the presence of Helicobacter species, particularly Helicobacter pylori and Helicobacter bilis, in the gallbladder of patients with non-malignant gallbladder diseases and control patients. PATIENTS AND METHODS: DNA was extracted from gallbladder samples from 122 consecutive patients undergoing cholecystectomy. The presence of Helicobacter genus-specific or Helicobacter pylori and Helicobacter bilis species-specific DNA was determined by polymerase chain reaction and sequence analysis. The presence of Helicobacter pylori-specific immunoglobulin G in the serum (n=84) and bile (n=104) samples was determined by enzyme linked immunosorbent assay. RESULTS: Helicobacter DNA was detected in 61 (50.0%) gallbladder samples: 29 of 60 (48.3%) patients with symptomatic gallstone, six of 10 (60.0%) patients with asymptomatic gallstones, 11 of 15 (73.3%) patients with other biliary diseases, and 15 of 37 (40.5%) control patients, respectively. Among them, 39 samples were positive for Helicobacter pylori but none were positive for Helicobacter bilis. Sequence analysis of Helicobacter genus-positive samples showed that 56 samples were Helicobacter pylori and five were Helicobacter species 'Liver 3' strain. Overall, there was no significant difference in the detection rate of Helicobacter DNA or the levels of serum and bile Helicobacter pylori-specific immunoglobulin G in the various biliary disease groups compared with control patients. Neither was there any significant difference in the blood biochemistry and liver function tests between patients with positive and negative Helicobacter DNA detection. CONCLUSION: Helicobacter species' DNA are commonly present in the gallbladder of patients with gallstone diseases and in controls, implying that Helicobacter infection alone may not play a significant role in the formation of gallstones. However, our results do not exclude the possibility of Helicobacter infection as a cofactor in the development of gallstones.     

Serum antibodies to enterohepatic Helicobacter spp. in patients with chronic liver diseases and in a population with high prevalence of H. pylori infection

BACKGROUND: Enteric Helicobacter species might be a risk factor for chronic liver and biliary tract diseases. AIMS: To analyse serum antibody levels to three enteric Helicobacter species in patients with various biliary tract and chronic liver diseases and compare results with corresponding parameters for an adult population group, known to have a high prevalence of Helicobacter pylori infection, and with healthy blood donors, to explore a possible association of enteric Helicobacter with chronic liver diseases. SUBJECTS: Sera of 90 patients with various chronic liver diseases, 121 Estonian adult persons and 68 blood donors were analysed. METHODS: Sera, previously tested for H. pylori were analysed for IgG to Helicobacter hepaticus, Helicobacter bilis and Helicobacter pullorum. ELISA was initially used for screening and exclusion of negative cases. Sera with positive ELISA results were further analysed by immunoblot. To remove cross-reactive antibodies between H. pylori and the enteric species, sera were pre-absorbed with lysed H. pylori cells. RESULTS: Liver patients showed a significantly higher seroprevalence to H. hepaticus and H. bilis, compared with the adult population group (p=0.0001 and 0.04, respectively), and to H. hepaticus, compared with blood donors (p=0.01). Patients with autoimmune hepatitis showed no significant antibody reactivity to the enteric Helicobacter spp. in contrast to patients with other chronic liver diseases. CONCLUSION: Patients with chronic liver diseases, except autoimmune hepatitis patients, showed increased antibody levels to H. bilis/H. hepaticus compared with the population and blood donors indicating a possible role of enteric Helicobacter in the natural course of chronic liver diseases. Immunoblot seems to be a promising method for serodiagnosis of infections with these fastidious pathogens.     

C-urea breath test in the management of Helicobacter pylori infection

The urea breath test is a noninvasive and very accurate test for the diagnosis of Helicobacter pylori infection. However, false negative urea breath test results have been reported to occur in a considerable percentage of the individuals taking proton pump inhibitors; the interval needed to be completely confident that false negative tests had been excluded has varied among the different studies between 6 and 14 days. The impact of H₂-receptor antagonists on the accuracy of urea breath test remains controversial, although, in contrast with proton pump inhibitors, the data suggest that H₂-receptor antagonists, for the most part, have little effect on the result of the urea breath test. The urea breath test does not represent a suitable tool for estimating the density of H. pylori colonization. The only quantitative information to be obtained from the urea breath test is that the higher the δ value, the lower the probability of a false-positive urea breath test result. Although some authors have demonstrated a correlation between urea breath test values and histological lesions of the gastric mucosa, the practical utility of this relationship remains unclear. It has been suggested that the pretreatment urea breath test has the potential to identify patients who require modification of the standard therapeutic regimen (for example, prolonging the duration of treatment or increasing the pharmacological dose when bacterial density is high), but other studies could not confirm this relationship. Some studies have shown that the urea breath test is less accurate in patients who have undergone partial gastrectomy. Finally, in contrast with biopsy-based methods, which are responsible for a high number of false-negative results when used to diagnose H. pylori infection in patients with upper gastrointestinal bleeding, urea breath test seems not to be negatively influenced by the presence of this complication.     

Helicobacter species and common gut bacterial DNA in gallbladder with cholecystitis

AIM:To analyze the association between Helicobacter spp. and some common gut bacteria in patients with cholecystitis. METHODS:A nested-polymerase chain reaction (PCR), specif ic to 16S rRNA of Helicobacter spp. was performed on paraff in-embedded gallbladder samples of 100 cholecystitis and 102 control cases. The samples were also analyzed for some common gut bacteria by PCR. Positive samples were sequenced for species identif ication. RESULTS: Helicobacter DNA was found in seven out of 100 cases of acute a...     

Use of bovine lactoferrin for Helicobacter pylori eradication

BACKGROUND: One-week triple therapy is the most frequently recommended treatment for Helicobacter pylori infection. Eradication rate is satisfactory, nevertheless is advisable to look for more effective therapies. AIM: To test the efficacy of a standard triple therapy plus bovine lactoferrin in the eradication of H. pylori infection. PATIENTS AND METHODS: One hundred and fifty consecutive H. pylori positive patients, suffering from dyspeptic symptoms were recruited in a 7-day triple therapy open randomised single centre study with rabeprazole, clarithromycin, tinidazole, bovine lactoferrin (group A) or rabeprazole, clarithromycin, tinidazole (group B), or a 10-day therapy with rabeprazole, clarithromycin, tinidazole (group C). H. pylori status was assessed 8 weeks after the end of the treatment by means of a 13C-urea breath test or a H. pylori stool antigen-test. RESULTS: Eradication rates (intention to treat/per protocol) were: group A (92.2/95.9%), group B (71.2/72.5%) and group C (70.2/75%). The efficacy of triple therapy added with lactoferrin was significantly higher than other two regimens (p=0.01, intention to treat analysis; p=0.005, per protocol analysis). CONCLUSION: These results suggest that lactoferrin tested in the present study was effective in curing H. pylori and could be a new agent to assist the antimicrobials in the eradication of the bacterium.     

New molecular marker for Trypanosoma (Duttonella) vivax identification

Trypanosoma vivax is a widespread hemoparasite in tropical areas and is pathogenic to ruminant domestic livestock as well as wild ruminants. The accurate identification of parasites in both hosts and vectors is crucial for epidemiological studies and disease control programs. We describe here the development of molecular markers specific for T. vivax identification. These markers were used to identify mouthpart infections in field-collected tsetse flies from Cameroon. The markers target the genomic sequence of a species-specific antigen from the bloodstream stages. No cross amplification with other trypanosome species was observed, which makes the markers a reliable tool to detect T. vivax infections, both in hosts and vectors. The PCR-amplified sequence contains a (CA)_n microsatellite repeat for which 11 different alleles were identified. This microsatellite, which showed high polymorphism, provides a suitable marker for population genetic studies.     

Isolation and characterization of Acanthamoeba spp. from air-conditioners in Kuala Lumpur, Malaysia

During a study on the quality of the indoor environment, Acanthamoeba spp. were detected in 20 out of 87 dust samples collected from air-conditioners installed in a four-story campus building located in Kuala Lumpur, Malaysia. Twenty-one cloned Acanthamoeba isolates designated as IMU1 to IMU21 were established from the positive primary cultures. Five species were identified from the 16 isolates according to the morphological criteria of Pussard and Pons; i.e. A. castellanii, A. culbertsoni, A. griffini, A. hatchetti and A. polyphaga. Species identities for the remaining five isolates (IMU4, IMU5, IMU15, IMU20 and IMU21), however, could not be determined morphologically. At genotypic characterization, these isolates were placed into T3 (IMU14); T5 (IMU16 and IMU17) and T4 (all the remaining isolates). To predict the potential pathogenicity of these Acanthamoeba isolates, thermo- and osmotolerance tests were employed; many isolates were predicted as potential human pathogens based on the outcome of these tests. This is the first time potentially pathogenic Acanthamoeba have been isolated from air-conditioners in Malaysia.     

Increased phospholipase activity in Helicobacter pylori strains isolated from patients with gastric carcinoma

PURPOSE: Phospholipase activity, one of Helicobacter pylori pathogenicity factors, has not been investigated enough, so far, although it may induce a remarkable damage to the gastric mucosa. In the present work, we have compared the whole phospholipase activity of H. pylori strains isolated from patients with gastric carcinoma with that of strains isolated from dyspeptic patients without gastric carcinoma. METHODS: We measured the phospholipase activity of one distinct H. pylori colony isolated from each of 10 patients with gastric carcinoma and 10 controls, dyspeptic patients without endoscopic and histological signs of gastric carcinoma. We also determined the phospholipase activity of 20 additional strains isolated from different areas of neoplastic and non-neoplastic tissue of two patients with gastric carcinoma, the cagA and vacA positive G27 and 328 wild strains and their respective vacA and cagA negative isogenic mutants. The whole phospholipase activity of strains was determined by measuring the release of (14)C-labeled palmitic acid from the radioactive l-3-phosphatidylcholine, 1,2-di[1-(14)C]palmiloyl substrate; results were expressed in pmol of palmitic acid per mg of protein. RESULTS: H. pylori strains isolated from patients with gastric carcinoma had levels of phospholipase activity significantly higher than those of strains isolated from controls (99.37 [S.D. 40.45] versus 34.46 [S.D. 16.46], P<0.001). In patients with gastric carcinoma, the mean phospholipase activity of strains isolated from neoplastic tissue was similar to that of strains isolated from non-neoplastic tissues (123.02 [S.D. 44.36] and 115.77 [S.D. 81.48], respectively. Interruption of cagA gene caused a ca. 20% reduction of phospholipase activity (36.38 versus 45.22 of the wild strain); that of vacA caused no reduction of phospholipase activity (26.53 and 25.37 of the wild strain). CONCLUSIONS: The infection by H. pylori strains that produce high levels of phospholipase may increase the risk of developing gastric carcinoma. We hypothesise that indirect products of phospholipase activity, such as prostaglandins, leukotrienes and lysophospholipids, may mediate carcinogenesis.     

Genetic analysis of ticks belonging to the Rhipicephalus sanguineus group in Latin America

Phylogenetic analyses based on mitochondrial 16S rDNA sequences were generated from Rhipicephalus sanguineus group specimens collected in 29 localities among 9 Latin-American countries, plus ticks collected in South Africa, Spain, and Italy. Sequences from Latin America generated six different haplotypes (A, B, C, D, E, and F). Phylogenetic analyses generated trees that segregated our tick sequences into two distinct clades: one is represented by haplotypes A–C, and South African R. sanguineus and Rhipicephalus turanicus ticks; the second clade is represented by haplotypes D–F, and European R. sanguineus and R. turanicus ticks. When haplotypes A–F are plotted in the Latin America map according to their geographical coordinates, it is clearly seen that haplotypes D–F are restricted to the southern portion of this continent, whereas haplotypes A–C are distributed in areas between northern Mexico and Brazil (except for the extreme south of this last country, where haplotype E was present). Hence, our phylogenetic analyses separated New World specimens of R. sanguineus into two distinct clades, one represented by tropical and subtropical populations (haplotypes A–C), here designated as the ‘tropical’ species. On the other hand, haplotypes D–F are here designated as the ‘temperate’ species because of their distribution in the southern portion of South America. Until recently, it was assumed that the R. sanguineus group was represented by a single species in the New World, namely R. sanguineus. While the present results coupled with recent studies support the presence of at least two species under the taxon R. sanguineus in the New World, they also show that even in the Old World, the taxon R. sanguineus might be represented by more than one species, since our phylogenetic analysis segregated European and South African R. sanguineus ticks into two distinct clades. The same can be applied for Spanish and South African R. turanicus.     

Helicobacter pylori diagnosis in patients with liver cirrhosis

BACKGROUND: In cirrhotics, Helicobacter pylori infection is the major cause of peptic lesions, which are an important cause of upper intestinal haemorrhage in these patients. However, some diagnostic methods are not accurate for H. pylori detection in cirrhotics. AIMS: The study assessed the accuracy of different diagnostic methods for H. pylori detection in cirrhotics with and without gastroduodenal lesions. METHODS: The study population comprised of 53 cirrhotics. All patients underwent upper endoscopy: three biopsies were taken in the antrum and three in the gastric body. Four biopsies were used for Giemsa staining, while two were used for a rapid urease test. A blood sample was obtained for serology using Western blotting, and a [13C]urea breath test was performed in all patients. Histological assessment was regarded as the gold standard for diagnosis of H. pylori infection. RESULTS: H. pylori infection was detected at histological assessment in 28 (52.8%) patients. The [13C]urea breath test, rapid urease test, and serology were positive in 27 (51%) patients, 23 (43.4%) patients, and 34 (64.1%) patients, respectively. Sensitivity and specificity were 92.9 and 96% for the [13C]urea breath test, 78.6 and 96% for the rapid urease test, and 78.6 and 52% for serology. CONCLUSIONS: The [13C]urea breath test is very accurate in cirrhotics, whilst both serology and the rapid urease test give disappointing results.     

Association of FCGR2A, JAK2 or HNF4A variants with ulcerative colitis in Koreans

Background

Recent genome-wide association studies have identified over 40 candidate genes contributing to ulcerative colitis susceptibility. The goal of this study was to test the reported ulcerative colitis susceptibility genes including FCGR2A, SLC26A3, JAK2 and HNF4A in Korean patients with ulcerative colitis and Crohn's disease.

Methods

Five single nucleotide polymorphisms from 4 loci including FCGR2A, SLC26A3, JAK2 and HNF4A were genotyped in 661patients with ulcerative colitis, 642 patients with Crohn's disease and 601 healthy controls.

Results

Statistically significant associations with ulcerative colitis were found at FCGR2A (rs1801274, p = 2.3 × 10⁻⁴, OR = 0.70 (95% CI = 0.57–0.84) under the allelic model), the JAK2 locus (rs10975003, p = 6.7 × 10⁻⁴, OR = 1.43 (95% CI = 1.16–1.77) under the allelic model) and HNF4A (rs6017342, p = 0.002, OR = 0.66 (95% CI = 0.51–0.85) under the allelic model). The association of FCGR2A was much stronger in female patients with ulcerative colitis (p = 5.7 × 10⁻⁶) than in males (p = 0.50). Except rs10975003 from the JAK2 locus, none showed positive association with Crohn's disease.

Conclusions

Our data suggest that FCGR2A, JAK2 or HNF4A variants play a role in the pathogenesis of ulcerative colitis in Koreans.     

Molecular characterization of the invasive Asian tiger mosquito, Aedes (Stegomyia) albopictus (Diptera: Culicidae) in Corsica

The Asian tiger mosquito Aedes albopictus, vector of various human viruses and parasites, has recently spread and established in many temperate regions including European countries. In the present study, we developed a simple PCR-based assay (the amplification of the internal transcribed spacer ITS2 within nuclear ribosomal rDNA) for molecular identification of A. albopictus and confirmed its presence in Corsica island. This assay may (i) facilitate future large scale studies and avoid misidentifications, especially because of the presence of co-occurring close species in this island and (ii) contribute to the monitoring of A. albopictus populations required for targeted control programs.     

Tumor necrosis factor-α in liver transplantation and resection: No evidence for a key role in ischemia-reperfusion injury

Experimental studies have shown that liver ischemia-reperfusion induces Kupffer cell activation and tumor necrosis factor-α (TNFα) release. The aim of this work was to determine whether severe hepatic ischemia and subsequent reperfusion triggers TNFα release in man. Serum TNFα was measured before and 3, 10, 30, 60, 120 min after revascularization and postoperatively at day 1 and 2 in 11 patients with orthotopic liver transplantation (group 1 and 4 patients with liver resection with vascular occlusion (group 2). In group 1, TNFα levels decreased during the first few minutes of reperfusion, then increased slightly to peak at 120 min (40 ± 13 pg/ml). Primary non-function occurred in 1 patient in whom low peroperative levels of TNFα levels were measured. In group 2, no significant changes in TNFα levels were observed. These data, in a small number of patients: (a) show that hepatic ischemia reperfusion does not result in major TNFα production; (b) do not support a primary pathogenic role for TNFα in damage after ischemia-reperfusion in humans.     

Inhibition of the growth of Plasmodium falciparum and Plasmodium berghei in vitro by an extract of Cochlospermum angolense (Welw.)

An extract of Cochlospermum angolense (Welw.) is used in the traditional medicine of Angola for the therapy of icterus and for the prophylaxis of malaria. From the roots of this plant red crystalline substances have been isolated and tested for their effect on Plasmodium falciparum in vitro and on the DNA and protein synthesis of Plasmodium berghei. The multiplication of P. falciparum was decreased to 50% of the control in the presence of 10 μg/ml extracted material and there was a total inhibition at a concentration of 50 μ/ml. If mice erythrocytes infected by P. berghei were incubated for 6 h with 25 μg/ml of the extract DNA synthesis was depressed to nearly background level. And, even more important, this effect could be demonstrated immediately. On the contrary, protein synthesis continued for at least 90 min at a reduced rate and stopped then. The results obtained show the direct antiparasitic effect of the substances extracted from C. angolense. The activity seems to be directed against DNA synthesis.     

Endoparasites of red fox (Vulpes vulpes) in the Slovak Republic with the emphasis on zoonotic species Echinococcus multilocularis and Trichinella spp.

Summary  Due to specific geographical localization, climatic and geomorphologic conditions, several serious parasitic diseases circulate in the territory of the Slovak Republic that makes this area an ideal model territory of the central European red fox system. The red fox is an important reservoir host of parasites, which can be spread to another animals and humans. Our study was aimed at determining the current prevalence of certain parasites in red foxes from the entire territory of the Slovak Republic and identifies some ecological factors influencing their epidemiology. Within the first systematic investigation of red foxes carried out between the years 2000 and 2006 in total 4026 foxes were examined for Echinococcus multilocularis (prevalence 31.1 %) and 4699 foxes were investigated for the presence of Trichinella spp. larvae (10.4 % infected). The results of the next separate study revealed that 83.3 % of 1198 red foxes in the Slovak Republic had coccidian oocysts and helminth eggs in their faeces. Fifteen helminth species including two trematode, four cestode and nine nematode species were detected by coprological examination. Nine of these parasite taxa have zoonotic potential: Capillaria spp. (prevalence 22.4 %), Ancylostoma caninum (18.1 %), Toxocara canis (12.5 %), Taenia spp. (12.2 %), Mesocestoides spp. (5.8 %), Strongyloides stercoralis (1.6 %), Hymenolepis diminuta (0.6 %), Dipylidium caninum (0.4 %) and Opisthorchis felineus (0.3 %). Toxascaris leonina was the most common helminth species found in this survey (42.9 %).     

Rifabutin based triple therapy for eradication of H. pylori primary and secondary resistant to tinidazole and clarithromycin

BACKGROUND: Rifabutin has been empirically used in Helicobacter pylori infections resistant to triple therapy. There are no data on primary and secondary resistance to rifabutin and its use in specific cases. AIM: To analyse the susceptibility and resistance to rifabutin in H. pylori-positive patients with or without previous H. pylori therapy and to test the efficacy of rifabutin in H. pylori resistant to clarithromycin and tinidazole. METHODS: Four hundred and twenty H. pylori-positive patients without previous exposure to triple therapy and 104 patients who had already received one course of triple therapy underwent upper endoscopy for dyspeptic symptoms and H. pylori susceptibility test. Amoxicillin, clarithromycin, tinidazole and rifabutin were evaluated for resistance and susceptibility. Forty patients with primary resistance to both clarithromycin and tinidazole and with susceptibility to amoxicillin and rifabutin, and 65 patients with secondary resistance and susceptibility to the same antibiotics were identified. All these patients received a 10-day triple therapy with pantoprazole amoxicillin and rifabutin. Treatment success was evaluated by the 13C-Urea Breath test. RESULTS: In naive patients 23% of strains were resistant to clarythromycin, 35% to tinidazole, 9% to both antibiotics, and none was resistant to rifabutin In patients already treated the percentages of resistant strains were 76, 64.4, 62.5 and 1%, respectively. With rifabutin based triple therapy eradication rates were (Per Protocol and Intention-to-Treat analysis) 100 and 87.5% in primary resistance to clarithromycin and tinidazole and 82.2 and 78.5% in secondary resistance. CONCLUSION: H. pylori primary and secondary resistances to clarithromycin and tinidazole are high in our geographic area, while resistance to rifabutin is rare. Rifabutin-based triple therapy, can be successfully used in primary and secondary resistance to clarithromycin and tinidazole according to the in vitro susceptibility test.     

Helicobacter pylori infection in patients with liver cirrhosis: facts and fictions

Helicobacter pylori infection could play a role in different clinical alterations observed in cirrhosis, from gastroduodenal lesions to hepatic encephalopathy. Although its prevalence in cirrhotics is similar to that in controls, H. pylori infection is responsible for the increased prevalence of peptic ulcer observed in these patients. The ammonia production by H. pylori urease does not seem to increase blood ammonia levels during cirrhosis, indicating that its role in hepatic encephalopathy could be marginalized in clinical practice. Dual and triple therapies have been shown to be equally effective for H. pylori eradication in these patients.     

Progression of Hypermethylation of the p16 INK4A Gene from Normal Liver to Nontumorous Liver and Hepatocellular Carcinoma: An Evaluation Using Quantitative PCR Analysis

The aim of this study was to determine to what extent hypermethylation of the p16 ^INK4A (p16) gene promoter is increased in nontumorous liver tissues compared with in normal liver, using two quantitative methylation-specific polymerase chain reaction (MS-PCR) methods and a bisulfite sequencing method. Methylation of the p16 gene was detected more frequently in nontumorous liver than in normal liver using the TaqMan PCR method. Methylation indices also were significantly higher in nontumorous than in normal liver. However, the bisulfite sequencing method did not detect significantly more methylation of the p16 gene in nontumorous than normal liver, nor was there a significant difference in the level of p16 mRNA. There may be a greater proportion of cells which contain methylated p16 in nontumorous than in normal liver. However, the difference was so small that the functional relevance to hepatocarcinogenesis remains elusive.     

Treatment of Helicobacter pylori infection. indications and regimens: an update

The management of Helicobacter pylori infection is still surrounded by controversy and uncertainties. Indications and correct application of current regimens for Helicobacter pylori infection are still considered a matter of debate. Regarding indications, only peptic ulcer and mucosa associated lymphoid tissue lymphoma are considered clear indications for treatment. In other conditions, such as atrophic gastritis, post gastric cancer resection, first-degree relatives of gastric cancer patients, dyspeptic patients, patients with gastro-oesophageal reflux disease and non-steroidal anti-inflammatory drug users, the value of Helicobacter pylori eradication is still controversial. The regimens for first-line and second-line treatment of Helicobacter pylori infection have been recommended by the Maastricht 2 Consensus Report. Although all the treatments are considered to be effective, physicians still do not agree on what first-line regimen should be used. Furthermore, a consensus on the duration of the antibiotic treatment is still lacking, although Maastricht guidelines for treatment of Helicobacter pylori infection recommend a one-week therapy. Also regimens, as a third-line treatment, and methods to improve compliance and clinical outcome are still a matter of debate. All these points will be considered in the present review.     

Evaluation of Five Probiotic Products for Label Claims by DNA Extraction and Polymerase Chain Reaction Analysis

Label claims were evaluated for five probiotic products. Specific oligonucleotide primers were designed for 11 species from the Bifidobacterium, Lactobacillus, and Streptococcus genera. Polymerase chain reaction, gel electrophoresis, and amplicon excision with DNA sequencing were performed: Sequence analysis and DNA homology comparisons followed. Bifidobacterium bifidum was not detected in two of the five samples by PCR analysis. Also, Lactobacillus species were found in two of the five product samples for which the species was not listed as an ingredient. We conclude that (1) lack of B. bifidum in two probiotic products may be attributed to different preparation standards among probiotic manufacturers, and (2) indentification of additional Lactobacillus species may represent contamination of the samples due to manufacturers utilizing shared equipment to produce all probiotics.