Patrolling Alveolar Macrophages Conceal Bacteria from the Immune System to Maintain Homeostasis.

During respiration, humans breathe in more than 10,000 liters of non-sterile air daily, allowing some pathogens access to alveoli. Interestingly, alveoli outnumber alveolar macrophages (AMs), which favors alveoli devoid of AMs. If AMs, like most tissue macrophages, are sessile, then this numerical advantage would be exploited by pathogens unless neutrophils from the blood stream intervened. However, this would translate to omnipresent persistent inflammation. Developing in vivo real-time intravital imaging of alveoli revealed AMs crawling in and between alveoli using the pores of Kohn. Importantly, these macrophages sensed, chemotaxed, and, with high efficiency, phagocytosed inhaled bacterial pathogens such as P. aeruginosa and S. aureus, cloaking the bacteria from neutrophils. Impairing AM chemotaxis toward bacteria induced superfluous neutrophil recruitment, leading to inappropriate inflammation and injury. In a disease context, influenza A virus infection impaired AM crawling via the type II interferon signaling pathway, and this greatly increased secondary bacterial co-infection.

A Functional Assessment of Fetal Liver and Monocyte-Derived Macrophages in the Lung Alveolar Environment.

It is becoming clear that every organ is seeded by a population of fetal liver-derived macrophages that are replaced at different rates by monocyte-derived macrophages. Using the Ms4a3tdTomato reporter mouse that reports on monocyte-derived alveolar macrophages (Mo-AMs) and our ability to examine AM function using our multichannel intravital microscopy, we examined the fetal-liver derived alveolar macrophage (FL-AM) and Mo-AM populations within the same mouse under various environmental conditions. The experiments unveiled that AMs migrated from alveolus to alveolus and phagocytosed bacteria identically regardless of ontogenic origin. Using 50 PFU of influenza A virus (IAV) determined using the Madin-Darby canine kidney (MDCK) cell line, we noted that both populations were susceptible to IAV-induced immunoparalysis, which also led to impaired phagocytosis of secondary bacterial infections. Both FL-AMs and Mo-AMs were trained by ß-glucan to resist IAV-induced paralysis. Over time (40 wk), Mo-AMs began to outperform FL-AMs, although both populations were still sensitive to IAV. Our data also show that clodronate depletion of AMs leads to replenishment, but by FL-AMs, and these macrophages do show some functional impairment for a limited time. Overall, the system is designed such that new macrophages rapidly assume the function of tissue-resident macrophages when both populations are examined in an identical environment. These data do differ from artificial depletion methods that compare Mo-AMs and FL-AMs.

FK506 impairs neutrophil migration that results in increased polymicrobial sepsis susceptibility.

OBJECTIVE: This study aimed to investigate the effects of FK506 on experimental sepsis immunopathology. It investigated the effect of FK506 on leukocyte recruitment to the site of infection, systemic cytokine production, and organ injury in mice with sepsis. METHODS: Using a murine cecal ligation and puncture (CLP) peritonitis model, the experiments were performed with wild-type (WT) mice and mice deficient in the gene Nfat1 (Nfat1-/-) in the C57BL/6 background. Animals were treated with 2.0 mg/kg of FK506, subcutaneously, 1 h before the sepsis model, twice a day (12 h/12 h). The number of bacteria colony forming units (CFU) was manually counted. The number of neutrophils in the lungs was estimated by the myeloperoxidase (MPO) assay. The expression of CXCR2 in neutrophils was determined using flow cytometry analysis. The expression of inflammatory cytokines in macrophage was determined using ELISA. The direct effect of FK506 on CXCR2 internalization was evaluated using HEK-293T cells after CXCL2 stimulation by the BRET method. RESULTS: FK506 treatment potentiated the failure of neutrophil migration into the peritoneal cavity, resulting in bacteremia and an exacerbated systemic inflammatory response, which led to higher organ damage and mortality rates. Failed neutrophil migration was associated with elevated CXCL2 chemokine plasma levels and lower expression of the CXCR2 receptor on circulating neutrophils compared with non-treated CLP-induced septic mice. FK506 did not directly affect CXCL2-induced CXCR2 internalization by transfected HEK-293 cells or mice neutrophils, despite increasing CXCL2 release by LPS-treated macrophages. Finally, the CLP-induced response of Nfat1-/- mice was similar to those observed in the Nfat1+/+ genotype, suggesting that the FK506 effect is not dependent on the NFAT1 pathway. CONCLUSION: Our data indicate that the increased susceptibility to infection of FK506-treated mice is associated with failed neutrophil migration due to the reduced membrane availability of CXCR2 receptors in response to exacerbated levels of circulating CXCL2.

Parental ancestry and risk of early pregnancy loss at high altitude.

High altitude pregnancy is associated with increased frequency of low birth weight infants and neonatal complications, the risks of which are higher in women of low-altitude ancestry. Does ancestry also influence the risk of miscarriage (pregnancy loss <20 weeks) in high-altitude pregnancy? To answer this, 5386 women from La Paz, Bolivia (3300-4150 m) with ≥1 live-born infant were identified. Data were extracted from medical records including maternal and paternal ancestry, demographic factors, and reproductive history. The risk of miscarriage by ancestry was assessed using multivariate logistic regression, adjusting for parity, and maternal age. Andean women experienced first live-births younger than Mestizo or European women (21.7 ± 4.6 vs 23.4 ± 8.0 vs 24.1 ± 5.1, P < .001). Andeans experienced more pregnancies per year of reproductive life (P < .001) and had significantly higher ratios of live-births to miscarriages than women of Mestizo or European ancestry (P < .001). Andean women were 24% less likely to have ever experienced a miscarriage compared to European women (OR:0.76; CI:0.62-0.90, P < .001). The woman's partner's ancestry wasn't a significant independent predictor of miscarriage. In conclusion, the risk of miscarriage at high altitude is lower in Andean women. The lack of a paternal ancestry effect suggests underlying mechanisms relate more to differential maternal adaptation in early pregnancy than fetal genetics.

Cutaneous temperature and pressure pain threshold in individuals with knee osteoarthritis.

INTRODUCTION: Osteoarthritis is a common disease in which skin temperature may be included among the pathophysiological factors. Thermography allows the mapping of cutaneous temperature and may be employed in the investigation of osteoarthritis. OBJECTIVES: To evaluate cutaneous temperature of the lower limbs, as well as to verify its association with pressure pain thresholds in individuals with knee osteoarthritis. MATERIAL AND METHODS: This case series study was conducted with individuals of both genders aged 48-77 years with unilateral knee osteoarthritis. Volunteers underwent thermographic evaluation by an infrared sensor (FLIR T650SC). The anterior region of the thigh and leg and the knee area were evaluated. Pressure pain thresholds (PPT) were evaluated by algometry (Pain Diagnostics, Great Neck, USA) at the vastus medialis, vastus lateralis, rectus femoris, and patellar tendon. Data analysis was conducted with the statistical package SPSS v.24 for Windows. Comparisons between affected and unaffected sides were made by paired Student's t-test or the Mann-Whitney U test, and associations between variables were assessed by Pearson or Spearman's correlation coefficient. In all cases, the significance level was set at p ≤ 0.05. RESULTS: Eleven volunteers (63.1 ±9.5 years) participated in this study. When comparing cutaneous temperature, only the region of the knee showed a significant difference between sides (p = 0.02). There were no differences between affected and unaffected knees regarding pain tolerance (PPT) at all sites evaluated. There were also no significant associations between the study variables. CONCLUSIONS: Individuals with knee osteoarthritis presented a higher temperature of the affected knee, but this was not associated with pressure pain thresholds.

Discordance between radiographic findings, pain, and superficial temperature in knee osteoarthritis.

INTRODUCTION: Knee pain is an imprecise marker of radiographic evidence of osteoarthritis (OA). These patients are more likely to develop central sensitisation to pain, a risk factor for chronic pain. OBJECTIVES: The aim of this study was to examine the associations among radiographic evidence of OA, self-reported pain, pressure pain thresholds (PPT), and superficial knee temperature in individuals with knee OA. MATERIAL AND METHODS: This cross-sectional study enrolled 25 patients with knee OA with clinic and radiographic diagnosis of bilateral OA (Kellgren-Lawrence grading scale 1-4 in X-ray images), but symptoms of pain and discomfort in only one of the knees, with pain perception in the symptomatic knee equal to or above 4 in a visual analogue scale. Volunteers underwent an evaluation including demographic data, superficial knee temperature (infrared thermography, mean superficial temperature of the region of the knee) and PPT (digital algometry at longus adductor, vastus lateralis, vastus medialis and tibialis anterior muscles, patellar tendon, and centre of the patella). RESULTS: Comparisons between symptomatic and asymptomatic knees revealed no differences regarding Kellgren-Lawrence classification, knee superficial temperature, or PPT. Significant weak and moderate associations were found between radiographic classification of OA and PPT of both knees in almost all sites evaluated. Nonetheless, superficial temperature of the knee was not associated with PPT or Kellgren-Lawrence grading scale. CONCLUSIONS: Patients with bilateral knee OA presented no differences in symptomatic and asymptomatic knees regarding radiographic evidence, knee temperature, and PPT, indicating that central sensitisation may be present in them. Radiographic classification of OA was significantly associated with PPT in both knees.

Phenotypic and genotypic profile of clinical and animal multidrug-resistant Salmonella enterica isolates from Mexico.

AIMS: The objective of this study was to obtain a phenotypic and genotypic profile of Salmonella enterica including multidrug-resistant (MDR) isolates from food-producing animals and clinical isolates, as well as their genetic relatedness in two different States of Mexico (Jalisco and State of Mexico). METHODS AND RESULTS: A total of 243 isolates were evaluated in terms of antimicrobial resistance (AMR) and related genes through a disk diffusion method and PCR respectively; we found 16 MDR isolates, all of them harbouring the blaCMY gene but not qnr genes, these isolates represent less than 10% of the collection. The pulsed-field gel electrophoresis revealed a higher genotypic similitude within isolates of State of Mexico than Jalisco. CONCLUSIONS: A low percentage of Salmonella isolates were resistant to relevant antibiotics in human health, nevertheless, the AMR and involved genes were similar despite the different serovars and origin of the isolates. SIGNIFICANCE AND IMPACT OF THE STUDY: This investigation provided an insight of the current status of AMR of Salmonella isolates in two States of Mexico and pinpoint the genes involved in AMR and their epidemiological relationship, the information could help to determine an adequate therapy in human and veterinary medicine.

Immunohistochemical identification of nail matrix melanocytes.

BACKGROUND: No previous studies have been conducted to determine the normal number of nail matrix melanocytes in Latin American individuals. The objective of this work was to determine the number of melanocytes per linear millimetre present in the nail matrix and the nail bed in samples obtained from Colombian individuals. METHODS: Twenty-six unilateral biopsies were taken from 19 cadavers subjected to clinical and medico-legal autopsies. These biopsy samples were processed with conventional histotechnology and immunohistochemistry (IHC) with anti-HMB-45 and anti-MiTF. Three sets of photographs (HE, HMB-45 and MiTF) were taken of each biopsy sample and independently assessed by three pathologists. Each observer counted the number of melanocytes present in 1 linear mm of the nail matrix or bed. RESULTS: We found an average of 4.6 melanocytes x linear mm with H & E staining, 9.8 with HMB-45 and 12.4 with MiTF. CONCLUSIONS: The use of IHC significantly increases and facilitates the identification of melanocytes in unilateral biopsies. Our IHC counts exceed the averages found in the literature. This finding warrants new studies to verify whether the Colombian population presents higher numbers of melanocytes in the nail matrix than other populations or whether the observed increase is a result of the use of MiTF.

Effect of marine by-product meals on hen egg production parameters, yolk lipid composition and sensory quality.

The effect of including 5% marine by-product meals in feeds of laying hens on egg production, composition and sensory characteristics was tested. Marine by-product meals were prepared using two methods: (i) cooking (100°C/10 min) followed by drying (60°C/24 hr) or (ii) grinding followed by drying. The raw materials used for meal production were scallop or squid viscera, shrimp heads or whole mackerel. A total of 108 laying hens were allocated to nine diet treatments; one control diet (corn and soya bean based) and eight experimental diets, containing 95% of the control feed and 5% of the experimental meal for three weeks. Daily intake was higher in hens fed the dried mackerel and cooked shrimp meals. All the experimental treatments showed significantly higher concentration of n-3 HUFA in yolk reserves and phospholipids compared to the control (0.12-0.13 g per 100 g), especially those with scallop or squid prepared by both methods (0.53-0.95 g per 100 g). Scallop, squid and shrimp meal inclusion in the feed produced eggs with more astaxanthin (0.22 mg per 100 g) while this carotenoid was absent in the control and mackerel treatments. Visual evaluation of raw yolk colour increased with the inclusion of marine by-product meals with higher values in hens fed shrimp heads (13), followed by scallop viscera (11), squid viscera (9), and with similar values for mackerel and control (4). The taste, aroma, texture and colour of cooked eggs from different treatments were not statically different when evaluated by a panel of 60 untrained people. These results suggest that meals from marine by-products are a better alternative for improving egg yolk composition by increasing n-3 HUFA when compared to fishmeal as they also increase astaxanthin and yolk pigmentation without affecting egg sensory characteristics.

Dental, periodontal and salivary conditions in diabetic children associated with metabolic control variables and nutritional plan adherence.

AIM: Diabetes mellitus is a chronic disease that has manifestations other than alterations in endocrine regulation or in metabolic pathways. Several diseases of the oral cavity have been associated with diabetes mellitus type 1 and 2 in young people according to their evolution. Scarce information exists regarding the role of diabetes and its association with the oral health status in paediatric diabetic patients. The aims of the study were to assess the quality of saliva, saliva acidogenicity, dental caries experience, fluorosis and periodontal status in diabetic patients and to evaluate their relationship with metabolic control variables and nutritional plan adherence. MATERIALS AND METHODS: The study population consisted of 60 paediatric patients with both types of diabetes mellitus. Saliva testing included stimulated flow, pH (using pH indicator strips), buffer capacity and Snyder's Test. DMFT/dmft and dental caries experience were determined on the basis of ICDAS II codes. The periodontal status was assessed by PI and GI and fluorosis by FI. Nutritional plan adherence was established from the subscale "Dietary Control" of the Diabetes Self-Management Profile questionnaire. Medical Data was retrieved from the clinical registers in the Diabetic Clinic. RESULTS: We describe the main characteristics of the oral cavity related variables of our population that might guide the clinical practice in similar settings; we found a dmft/DMFT of 1.71 ± 1.74 and 0.64 ± 1.03, PI of 1.91 ± 0.75, GI of 0.50 ± 0.56 and a fluorosis prevalence of 61%. We identified several correlated variables, which indicate strong associations between the nutritional habits of the patients and co-occurrence of oral cavity physiopathological alterations. Several correlations were found between acidogenic activity of the saliva (Snyder Test) and the percentage of adherence to the nutritional plan and to the dmft index. Furthermore, a significant correlation between the buffering capacity of the saliva and the glycemic control of the participants was found. Neither an association nor a difference among means was found between treatment regime and the plaque index. CONCLUSION: The results of the present study concluded that there was a significant relationship between diabetes mellitus and an increased prevalence of oral cavity related diseases in the paediatric population. These are also associated with a poor adherence to the nutritional plan.

Identification of a Hemagglutinin from Gallibacterium anatis.

Gallibacterium anatis has the ability to hemagglutinate rabbit erythrocytes; however, no bacterial component has yet been associated with this function. In the present work, a protein of approximately 65 kDa with hemagglutinating activity for glutaraldehyde-fixed chicken erythrocytes was purified by ion interchange chromatography from G. anatis F149(T) secreted proteins. The protein was recognized by a rabbit polyclonal serum against a hemagglutinin from Avibacterium paragallinarum. The 65 kDa purified protein presented identity with a G. anatis filamentous hemagglutinin by mass spectrometric analysis. As well, the bacterial surface of G. anatis was labeled by immune gold assays using a polyclonal serum against the 65-kDa protein. A similar protein was recognized in four other G. anatis strains by immunoblots using the same antiserum. The protein binds sheep or pig biotinylated fibrinogen, suggesting an interaction with basement membrane eukaryotic cells components, and the protein is present in G. anatis biofilms. Overall, the results suggest that the 65 kDa hemagglutinin is a common antigen and a potential virulence factor in G. anatis.

Antibacterial effect of roselle extracts (Hibiscus sabadariffa), sodium hypochlorite and acetic acid against multidrug-resistant Salmonella strains isolated from tomatoes.

UNLABELLED: Antibiotic-resistant Salmonella strains were isolated from saladette and red round type tomatoes, and an analysis done of the antibacterial activity of roselle calyx extracts against any of the identified strains. One hundred saladette tomato samples and 100 red round tomato samples were collected from public markets. Each sample consisted of four whole tomatoes. Salmonella was isolated from the samples by conventional culture procedure. Susceptibility to 16 antibiotics was tested for the isolated Salmonella strains by standard test. The antibacterial effect of four roselle calyx extracts (water, methanol, acetone and ethyl acetate), sodium hypochlorite and acetic acid against antibiotic-resistant Salmonella isolates was evaluated on contaminated tomatoes. Twenty-four Salmonella strains were isolated from 12% of each tomato type. Identified Salmonella serotypes were Typhimurium and Typhi. All isolated strains exhibited resistance to at least three antibiotics and some to as many as 12. Over contaminated tomatoes, the roselle calyx extracts produced a greater reduction (2-2·6 log) in antibiotic-resistant Salmonella strain concentration than sodium hypochlorite and acetic acid. SIGNIFICANCE AND IMPACT OF THE STUDY: The presence of multidrug-resistant Salmonella in vegetables is a significant public health concern. Multidrug-resistant Salmonella strains were isolated from raw tomatoes purchased in public markets in Mexico and challenged with roselle Hibiscus sabdariffa calyx extracts, sodium hypochlorite and acetic acid. On tomatoes, the extracts caused a greater reduction in the concentration of antibiotic-resistant Salmonella strains than sodium hypochlorite and acetic acid. Roselle calyx extracts are a potentially useful addition to disinfection procedures of raw tomatoes in the field, processing plants, restaurants and homes.

Protection Conferred by Infectious Coryza Vaccines Against Emergent Avibacterium paragallinarum Serovar C-1.

Infectious coryza is an upper respiratory disease of chickens caused by Avibacterium paragallinarum. Outbreaks of infectious coryza caused by Av. paragallinarum serovar C-1 isolates in coryza-vaccinated flocks in Ecuador and Mexico have been reported. In the current study, the protection conferred by four commercially available, trivalent infectious coryza vaccines in chickens challenged with a serovar C-1 isolate from an apparent coryza vaccine failure in a layer flock in Mexico was evaluated. Only one infectious coryza vaccine provided a good protection level (83%) in vaccinated chickens. These results might explain the infectious coryza outbreaks in vaccinated flocks that have been observed in the field.

An intermediate respiratory care unit for spinal cord-injured patients. A retrospective study.

OBJECTIVES: Patients with cervical spinal cord injury (SCI) may need prolonged mechanical ventilation (MV) and a long stay in the Intensive Care Unit. An intermediate respiratory care unit (IRCU) can shorten that stay, optimizing hospital resources. The aim of our work has been to evaluate the activity of such a unit in our hospital. METHODS: This is a descriptive retrospective study based on the data of patients with SCI and respiratory failure discharged from our IRCU between 1 July 2010 and 28 February 2013. RESULTS: We have analysed data from 146 patients with SCI, adding up to 228 admissions (68 first admissions and 160 readmissions due to complications or scheduled review visits). Sixty-three out of the 68 newly admitted patients survived their first admission (92.6%). Length of hospitalization was 195.6±110.4 days, 22 were admitted to monitor their respiratory status and 46 were on MV on admission. Of these, 26 (38.2%) were admitted to attempt weaning from the respirator and 20 (29.4%) to enter a programme of permanent respiratory support. Weaning was successful in 23 out of 26 patients (88.4%), the process taking 47.2±49.3 days. Forty of them (58.8%) were discharged to their home. CONCLUSIONS: An IRCU can manage a substantial number of severe SCI patients who need MV, and an important number of them can be weaned from the respirator. It may also achieve a good success rate in the integration of MV-dependent patients within family and society.

Genotyping, pathogenicity, and immunogenicity of Avibacterium paragallinarum serovar B-1 isolates from the Americas.

The bacterium Avibacterium paragallinarum is the etiologic agent of infectious coryza of chickens. Among the nine Kume serovars currently recognized in this bacterium, serovar B-1 is a common serovar in the Americas. In the current study, serovar B-1 isolates from Ecuador (seven isolates), Mexico (seven isolates) and Panama (two isolates) were genotyped. In addition one Panamanian, one Ecuadorian, and two Mexican isolates were used in a vaccination-challenge trial in which the vaccine was based on the 2671 serovar B-1 reference strain. Genotyping by enterobacterial repetitive intergenic consensus-based PCR (ERIC-PCR) resulted in ten distinguishable ERIC patterns for the 16 isolates and the two reference strains of Av. paragallinarum included in the study. No ERIC patterns were shared among isolates of the three different countries. In the vaccination-challenge trial, one isolate from Panama showed a significantly lower virulence than did the three other isolates. In terms of cross-protection, chickens vaccinated with reference strain 2671 and challenged with an Ecuadorian strain showed 40% protection, a significantly lower protection than the homologous protection level. The other three field isolates gave a similar protection level to the homologous challenge.

Phylogenetic relationship of serovar C-1 isolates of Avibacterium paragallinarum.

The bacterium Avibacterium paragallinarum is the etiologic agent of infectious coryza of chickens. Serovar C-1 has emerged in infectious coryza outbreaks in layer hens of Ecuador and Mexico. In the current study, genotyping and phylogenetic analyses of five Ecuadorian and 10 Mexican isolates of Av. paragallinarum serovar C-1 were performed. All 15 isolates share a unique enterobacterial repetitive intergenic consensus-based-PCR fingerprint and have identical 16S ribosomal RNA and hemagglutinin antigen gene sequences. Results indicate that Ecuadorian and Mexican isolates of serovar C-1 of Av. paragallinarum have a clonal relationship.

Frequency of indicator bacteria, Salmonella and diarrhoeagenic Escherichia coli pathotypes on ready-to-eat cooked vegetable salads from Mexican restaurants.

UNLABELLED: The presence of coliform bacteria, faecal coliforms, Escherichia coli, diarrhoeagenic E. coli pathotypes (DEP) and Salmonella were determined in ready-to-eat cooked vegetable salads (RECS) from restaurants in Pachuca city, Mexico. The RECS were purchased from three types of restaurants: national chain restaurants (A), local restaurants (B) and small restaurants (C). Two restaurants for each A and B, and three for C, were included. Forty RECS samples were purchased at each A and B restaurant and 20 at each C restaurant. Of the overall total of 220 analysed samples, 100, 98·2, 72·3, 4·1 and 4·1% had coliform bacteria, faecal coliforms, E. coli, DEP and Salmonella, respectively. Identified DEP included enteropathogenic E. coli (EPEC), enterotoxigenic E. coli (ETEC) and Shiga toxin-producing E. coli (STEC). The EPEC, ETEC and STEC were isolated each from 1·4% of samples. No E. coli O157:H7 were detected in any STEC-positive samples. The analysis of Kruskal-Wallis anova and median test of microbiological data showed that the microbiological quality of RECS did not differ between the different restaurants (P > 0·05). SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report regarding microbiological quality and Salmonella, enteropathogenic E. coli (EPEC), enterotoxigenic E. coli (ETEC) and Shiga toxin-producing E. coli (STEC) isolation from ready-to-eat cooked vegetable salads from Mexican restaurants. Ready-to-eat cooked vegetable salads could be an important factor contributing to the endemicity of EPEC, ETEC and STEC, and Salmonella caused gastroenteritis in Mexico.

Presence of indicator bacteria, diarrhoeagenic Escherichia coli pathotypes and Salmonella in fresh carrot juice from Mexican restaurants.

UNLABELLED: Coliform bacteria (CB), faecal coliforms (FC), Escherichia coli, diarrhoeagenic E. coli pathotypes (DEP) and Salmonella frequencies were determined for fresh carrot juice from restaurants in Pachuca city, Mexico. Two hundred and eighty carrot juice samples were purchased in three types of restaurants: (A), national chain restaurants; (B), local restaurants; and (C), very small restaurants. Two restaurants for each A and B, and three for C, were included. Forty juice samples were purchased at each restaurant. All tested juice samples had poor microbiological quality. Of these samples, 100, 96·8, 54·3, 8·9 and 8·6% had CB, FC, E. coli, DEP and Salmonella, respectively. CB were present in all juice samples regardless of source, with limits ranging from 3·6 × 10² to 8·5 × 107 CFU ml⁻¹, and the limits for FC and E. coli were <3 to 1100 MPN ml⁻¹ and <3 to 460 MPN, respectively. DEP and Salmonella were isolated from samples from all the restaurants at levels of 5% or above: DEP, 5% (A1, B2, 10% (A2, B1, C1, C2) and 12·5% (C3); Salmonella, 5% (A1, A2, B2), 7·5% (C2), 10% (C1, 12·5% (B1) and 15% (C3). SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report of microbiological quality and Salmonella, enteroinvasive E. coli (EIEC), enterotoxigenic E. coli (ETEC) and Shiga toxin-producing E. coli (STEC) isolation from fresh carrot juice in Mexico. Fresh carrot juice from restaurants could be an important factor contributing to the endemicity of EIEC-, ETEC- and STEC- and Salmonella-caused gastroenteritis in Mexico.