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Glutathione peroxidase 3 (GPx3) is the only extracellular selenoprotein (Sel) that enzymatically reduces H 2 O 2 to H 2 O and O 2 . Two GPx3 ( CqGPx3 ) cDNAs were characterized from crayfish Cherax quadricarinatus . The nerve cord CqGPx3a isoform encodes for a preprotein containing an N-terminal signal peptide of 32 amino acid residues, with the mature Sel region of 192 residues and a dispensable phosphorylation domain of 36 residues. In contrast, the pereiopods CqGPx3b codes for a precursor protein with 19 residues in the N-terminal signal peptide, then the mature 184 amino acid residues protein and finally a Pro-rich peptide of 42 residues. CqGPx3 are expressed in cerebral ganglia, pereiopods and nerve cord. CqGPx3a is expressed mainly in cerebral ganglia, antennulae and nerve cord, while CqGPx3b was detected mainly in pereiopods. CqGPx3a expression increases with high temperature and hypoxia; meanwhile, CqGPx3b is not affected. We report the presence and differential expression of GPx3 isoforms in crustacean tissues in normal conditions and under stress for high temperature and hypoxia. The two isoforms are tissue specific and condition specific, which could indicate an important role of CqGPx3a in the central nervous system and CqGPx3b in exposed tissues, both involved in different responses to environmental stressors.

Equine infectious anemia (EIA) is a highly infectious disease in members of the Equidae family, caused by equine infectious anemia virus (EIAV). The disease severity ranges from subclinical to acute or chronic, and causes significant economic losses in the equine industry worldwide. Serologic tests for detection of EIAV infection have some concerns given the prolonged seroconversion time. Therefore, molecular methods are needed to improve surveillance programs for this disease. We attempted detection of EIAV in 6 clinical and 42 non-clinical horses in Nuevo Leon State, Mexico, using the agar gel immunodiffusion (AGID) test for antibody detection, and nested and hemi-nested PCR for detection of proviral DNA. We found that 6 of 6, 5 of 6, and 6 of 6 clinical horses were positive by AGID, nested PCR, and hemi-nested PCR, respectively, whereas 0 of 42, 1 of 42, and 9 of 42 non-clinical horses were positive by these tests, respectively. BLAST analysis of the 203-bp 5'-LTR/ tat segment of PCR product revealed 83-93% identity with EIAV isolates in GenBank and reference strains from other countries. By phylogenetic analysis, our Mexican samples were grouped in a different clade than other sequences reported worldwide, indicating that the LRT/ tat region represents an important target for the detection of non-clinical horses.

In northern Mexico, the distilled spirit sotol with a denomination of origin is made from species of Dasylirion . The configuration of the volatile metabolites produced during the spontaneous fermentation of Dasylirion sp. must is insufficiently understood. In this study, the aim was to investigate the composition of the microbial consortia, describe the variation of volatile metabolites, and relate such profiles with their particular flavor attributes during the fermentation of sotol ( Dasylirion sp.) must. Ascomycota was the phylum of most strains identified with 75% of total abundance. The genus of fermenting yeasts constituted of 101 Pichia strains and 13 Saccharomyces strains. A total of 57 volatile metabolites were identified and grouped into ten classes. The first stage of fermentation was composed of diesel, green, fruity, and cheesy attributes due to butyl 2-methylpropanoate, octan-1-ol, ethyl octanoate, and butanal, respectively, followed by a variation to pungent and sweet descriptors due to 3-methylbutan-1-ol and butyl 2-methylpropanoate. The final stage was described by floral, ethereal-winey, and vinegar attributes related to ethyl ethanimidate, 2-methylpropan-1-ol, and 2-hydroxyacetic acid. Our results improve the knowledge of the variations of volatile metabolites during the fermentation of sotol must and their contribution to its distinctive flavor.

Macro- and microorganism activities are important for the effectiveness of the slow sand filtration (SSF), where native microorganisms remove contaminants mainly by substrate competition, predation, and antagonism. The aim of the present study was to evaluate the addition of the oligodynamic metals iron, copper, and brass, inserted separately into SSF to enhance pollutant removal in water samples. Four laboratory-scale SSFs were built and tested: control, iron, copper, and brass. Water analysis included physicochemical evaluation, total and fecal coliform quantification. An analysis on microbial communities in the SSFs schmutzdecke was achieved by using 16S rRNA amplification, the Illumina MiSeq platform, and the QIIME bioinformatics software. The results demonstrated that inorganic and organic contaminants such as coliforms were removed up to 90%. The addition of metals had no significant effect (p > 0.05) on the other parameters. The microbial community analysis demonstrated different compositions of the SSF with brass-influent, where the eukaryote Streptophyta was predominant (31.4%), followed by the acetic acid bacteria Gluconobacter (24.6%), and Acetobacteraceae (7.7%), these genera were absent in the other SSF treatments. In conclusion, the use of a SSF system can be a low cost alternative to reduce microbial contamination in water and thus reduce gastrointestinal diseases in rural areas.

Pollution of freshwater ecosystems from polycyclic aromatic hydrocarbons (PAHs) is a global concern. The US Environmental Protection Agency (EPA) has included the PAHs pyrene, phenanthrene, and naphthalene among the 16 priority compounds of special concern for their toxicological effects. The aim of this study was to adapt and characterize a microbial consortium from ore waste with the potential to remove these three PAHs from water. This microbial consortium was exposed to the target PAHs at levels of 5, 10, 20, 50, and 100 mg L −1 for 14 days. PAH bioremoval was measured using the analytical technique of solid phase microextraction, followed by gas chromatography mass spectrometry (SPME-GC/MS). The results revealed that up to 90% of the target PAHs can be removed from water after 14 days at a concentration level of 100 mg L −1 . The predominant group of microorganisms identified at the phylum taxonomic level were the Proteobacteria, while the Actinobacteria were the predominant subgroup. The removal of phenanthrene, naphthalene, and pyrene predominantly occurred in specimens of genera Stenotrophomonas , Williamsia , and Chitinophagaceae, respectively. This study demonstrates that the use of specific microorganisms is an alternative method of reducing PAH levels in water.

It is estimated that 15% of all newborns admitted to the neonatal intensive care unit (NICU) for suspected sepsis receive multiple broad-spectrum antibiotics without pathogen identification. The gold standard for bacterial sepsis detection is blood culture, but the sensitivity of this method is very low. Recently, amplification and analysis of the 16S ribosomal DNA (rDNA) bacterial gene in combination with denaturing gradient gel electrophoresis (DGGE) has proven to be a useful approach for identifying bacteria that are difficult to isolate by standard culture methods. The main goal of this study was to compare two methods used to identify bacteria associated with neonatal sepsis: blood culture and broad range 16S rDNA-DGGE. Twenty-two blood samples were obtained from newborns with (n = 15) or without (n = 7) signs and symptoms of sepsis. Blood samples were screened to identify pathogenic bacteria with two different methods: (1) bacteriological culture and (2) amplification of the variable V3 region of 16S rDNA-DGGE. Blood culture analysis was positive in 40%, whereas 16S rDNA-DGGE was positive in 87% of neonatal sepsis cases. All 16S rDNA-DGGE positive samples were associated with some other signs of neonatal sepsis., Conclusion: Our study shows that the molecular approach with 16S rDNA-DGGE identifies twofold more pathogenic bacteria than bacteriological culture, including complex bacterial communities associated with the development of bacterial sepsis in neonates. What is Known: • Neonatal sepsis affects 2.3% of birth in the NICU with a high mortality risk. • Evidence supports the use of molecular methods as an alternative to blood culture for identification of bacterial associated neonatal sepsis. What is New: • The DGGE gel is a good methodological approach for the identification of bacterial in neonatal blood samples. • This study describes the pattern of electrophoretic mobility obtained by DGGE gels and allows to determine the type of bacteria associated in the development of neonatal sepsis.

Reclaimed wastewater for irrigation is an opportunity for recovery of this natural resource. In this study, microbial risk from the use of treated wastewater for irrigation of recreational parks in the city of Chihuahua, evaluating the effect of distribution distance, season, and presence of storage tanks, was analyzed. Escherichia coli , Salmonella , and multidrug-resistant bacteria were recovered from samples of reclaimed water and soils at recreational parks in Chihuahua by the membrane filtration method, using selected agars for microbial growth. Samples were taken at three different seasons. No correlation in the presence of microbial indicators and multidrug-resistant bacteria ( p > 0.05) was found between the distance from the wastewater treatment plant to the point of use. Presence of storage tanks in parks showed a significant effect ( p < 0.05) with a higher level of E. coli . The highest count in wastewater occurred in summer. We isolated 392 multidrug-resistant bacteria from water and soil; cluster analysis showed that the microorganisms at each location were of different origins. Irrigation with reclaimed wastewater did not have a negative effect on the presence of microbial indicators of the quality of soils in the parks. However, the prevalence of multidrug-resistant bacteria still represents a potential risk factor for human health., Competing Interests: The authors declare no conflict of interest.

In areas lacking potable water treatment, drinking contaminated water may represent a public health threat. In addition to enteropathogenic bacteria and parasites, fecal contamination in water environments is associated with the transmission of enteric viruses and other causal agents of infectious disease. Rotavirus and norovirus are the main enteric viral agents responsible for diarrheic outbreaks. The aim of the present study was to detect seasonal variation of rotavirus and norovirus in the surface water at Bassaseachic Falls National Park during 2013. Rivers and streams within and nearby this park were sampled once in each season during 2013. Viral concentration was carried out by a handmade filtration equipment, using a commercial electropositive membrane coupled with the virus absortion elution technique (VIRADEL©). Detection of rotavirus and norovirus was performed by SYBR Green reverse transcription-real time polymerase chain reaction (SYBR GREEN© RT-qPCR) analyses. Norovirus genogroup II was detected in samples collected in June and October 2013. In the case of rotavirus, genogroup A was detected in March and June. The presence of rotavirus and norovirus was related to viral acute diarrhea in children less than five years of age, who were inhabiting the sampled areas. This may indicates that the contaminated water was potentially a risk factor for regional diarrheic outbreaks.

The availability of good quality water resources is essential to ensure healthy crops and livestock. The objective of this study was to evaluate the level of pollution in Bustillos Lagoon in northern Mexico. Physical-chemical parameters like sodium, chloride, sulfate, electrical conductivity, nitrates, and the pesticide dichlorodiphenyltrichloroethane (DDT) were analyzed to determine the water quality available in the lagoon. Although DDT has been banned in several countries, it is still used for agricultural purposes in Mexico and its presence in this area had not been analyzed previously. Bustillos Lagoon was divided into three zones for the evaluation: (1) industrial; (2) communal lands; and (3) agricultural. The highest concentrations of sodium (2360 mg/L) and SAR (41 meq/L) reported in the industrial zone are values exceeding the United Nations Food and Agricultural Organization (FAO) irrigation water quality guidelines. DDT and its metabolites were detected in all of the 21 sites analyzed, in the agricultural zone ∑DDTs = 2804 ng/mL, this level is much higher than those reported for other water bodies in Mexico and around the world where DDT has been used heavily. The water in the communal zone is the least contaminated, but can only be recommended for irrigation of plants with high stress tolerance and not for crops.

Bacterial pathogens are a leading cause of waterborne disease, and may result in gastrointestinal outbreaks worldwide. Inhabitants of the Bassaseachic Falls National Park in Chihuahua, Mexico show seasonal gastroenteritis problems. This aim of this study was to detect enteropathogenic microorganisms responsible for diarrheal outbreaks in this area. In 2013, 49 surface water samples from 13 selected sampling sites along the Basaseachi waterfall and its main rivers, were collected during the spring, summer, autumn, and winter seasons. Fecal and total coliform counts were determined using standard methods; the AutoScan-4 system was used for identification of isolates and the antibiotic resistance profile by challenging each organism using 21 antibiotics. Significant differences among seasons were detected, where autumn samples resulted in the highest total (p < 0.05) and fecal (p < 0.001) coliform counts, whereas the lowest total coliform counts were recorded in spring. Significant differences between sampling sites were observed, where samples from sites 6, 8, and 11 had the highest total coliform counts (p < 0.009), whereas samples from site 9 exhibited the lowest one. From the microbiological analysis, 33 bacterial isolates from 13 different sites and four sampling seasons were selected; 53% of isolates were resistant to at least one antibiotic, and 15% exhibited a multidrug resistance (MDB) phenotype. MDB were identified as Klebsiella oxytoca (two out of four identified isolates), Escherichia coli (2/7), and Enterobacter cloacae (1/3). In addition, some water-borne microorganisms exhibited resistance to cefazoline, cefuroxime, ampicillin, and ampicillin-sulbactam. The presence of these microorganisms near rural settlements suggests that wastewater is the contamination source, providing one possible transmission mechanism for diarrheal outbreaks.

Assimilatory nitrate reduction (ANR) is a pathway wherein NO(3)(-) is reduced to NH(4)(+), an N species that can be incorporated into the biomass. There is little information about the ANR genes in Archaea and most of the known information has been obtained from cultivable species. In this study, the diversity of the haloarchaeal assimilatory nitrate-reducing community was studied in an extreme saline alkaline soil of the former lake Texcoco (Mexico). Genes coding for the assimilatory nitrate reductase (narB) and the assimilatory nitrite reductase (nirA) were used as functional markers. Primers to amplify and detect partial narB and nirA were designed. The analysis of these amplicons by cloning and sequencing showed that the deduced protein fragments shared >45% identity with other NarB and NirA proteins from Euryarchaeota and <38% identity with other nitrate reductases from Bacteria and Crenarchaeota. Furthermore, these clone sequences were clustered within the class Halobacteria with strong support values in both constructed dendrograms, confirming that desired PCR products were obtained. The metabolic capacity to assimilate nitrate by these haloarchaea seems to be important given that at pH 10 and higher, NH(4)(+) is mostly converted to toxic and volatile NH(3), and NO(3)(-) becomes the preferable N source.

Flooding an extreme alkaline-saline soil decreased alkalinity and salinity, which will change the bacterial populations. Bacterial 16S rDNA libraries were generated of three soils with different electrolytic conductivity (EC), i.e. soil with EC 1.7 dS m(-1) and pH 7.80 (LOW soil), with EC 56 dS m(-1) and pH 10.11 (MEDIUM soil) and with EC 159 dS m(-1) and pH 10.02 (HIGH soil), using universal bacterial oligonucleotide primers, and 463 clone 16S rDNA sequences were analyzed phylogenetically. Library proportions and clone identification of the phyla Proteobacteria, Actinobacteria, Acidobacteria, Cyanobacteria, Bacteroidetes, Firmicutes and Cloroflexi showed that the bacterial communities were different. Species and genera of the Rhizobiales, Rhodobacterales and Xanthomonadales orders of the alpha- and gamma-subdivision of Proteobacteria were found at the three sites. Species and genera of the Rhodospirillales, Sphingobacteriales, Clostridiales, Oscillatoriales and Caldilineales were found only in the HIGH soil, Sphingomonadales, Burkholderiales and Pseudomonadales in the MEDIUM soil, Myxococcales in the LOW soil, and Actinomycetales in the MEDIUM and LOW soils. It was found that the largest diversity at the order and species level was found in the MEDIUM soil as bacteria of both the HIGH and LOW soils were found in it.

Triatoma dimidiata is the only reported Chagas disease vector in Campeche, Mexico. The purpose of this study was to determine the genetic variability of vectors from Campeche coastal and rain forest areas and establish a phylogenetic relationship with other T. dimidiata populations by analyzing the internal transcribed spacer-2 (ITS-2) region. The sequence length of samples from Campeche ranged from 469 to 478 basepairs. The ITS-2 variability among the populations enabled us to classify them into two clades with an 18-22 nucleotide difference. The genetic distance (0.042) between them confirms this divergence. Phylogenetic analysis of gene genealogies confirmed these two clades. Furthermore, the population genetic analyses showed two groups with little genetic similarity or migration between them. One group was associated with the tropical forest area and the other group was associated with a mainly coastal distribution. This correlation was also observed when T. dimidiata from other regions of Mexico and Central America were analyzed.

The aim of this study was to assess the bacterial diversity associated with a corrosive biofilm in a steel pipeline from the Gulf of Mexico used to inject marine water into the oil reservoir. Several aerobic and heterotrophic bacteria were isolated and identified by 16S rRNA gene sequence analysis. Metagenomic DNA was also extracted to perform a denaturing gradient gel electrophoresis analysis of ribosomal genes and to construct a 16S rRNA gene metagenomic library. Denaturing gradient gel electrophoresis profiles and ribosomal libraries exhibited a limited bacterial diversity. Most of the species detected in the ribosomal library or isolated from the pipeline were assigned to Proteobacteria (Halomonas spp., Idiomarina spp., Marinobacter aquaeolei, Thalassospira sp., Silicibacter sp. and Chromohalobacter sp.) and Bacilli (Bacillus spp. and Exiguobacterium spp.). This is the first report that associates some of these bacteria with a corrosive biofilm. It is relevant that no sulfate-reducing bacteria were isolated or detected by a PCR-based method. The diversity and relative abundance of bacteria from water pipeline biofilms may contribute to an understanding of the complexity and mechanisms of metal corrosion during marine water injection in oil secondary recovery.

The infection frequency associated to bacterial conjunctivitis, corneal ulcers (CU), and endophthalmitis was studied along a five years period. The isolation and identification of microorganisms were performed by culture-based methods and biochemical test respectively. Also, a nested PCR to detect gram-negative and gram-positive bacteria in the clinical samples was assayed. Nested PCR was a more efficient method than culture to detect bacteria in the samples. The most frequently isolated species was Staphylococcus epidermidis, a bacterium commonly considered as a human saprophyte. The S. epidermidis strains from conjunctivitis, CU, and endophthalmitis exhibited 46, 33.9, and 34.1% of oxacilin-resistance respectively. A total of 28% of intermediate-vancomycin resistance (MIC = 8-16 microg/ml) was observed among S. epidermidis strain collection. The UPGMA cluster analysis of the multiresistance profile data of intermediate vancomycin-resistant S. epidermidis strains showed a high phenotypic diversity and no relationship between each group and their clinical origin. The biofilm formation capacity was broadly distributed (66%), particularly among intermediate-vancomycin strains (> 75%). In brief, S. epidermidis displayed a high diversity of antibiotic resistance profiles and biofilm formation capacity. These phenotypic traits could explain the high isolation frequency of S. epidermidis from ocular infections and oblige to review the saprophytic status of these bacteria.

Sixteen co-cultures composed of four bacteria and four fungi grown on sugarcane bagasse pith were tested for phenanthrene degradation in soil. The four bacteria were identified as Pseudomonas aeruginose, Ralstonia pickettii, Pseudomonas sp. and Pseudomonas cepacea. The four fungi were identified as: Penicillium sp., Trichoderma viride, Alternaria tenuis and Aspergillus terrus that were previously isolated from different hydrocarbon-contaminated soils. Fungi had a statistically significant positive (0.0001

The hemophagocytic syndrome is a reactive disorder of the mononuclear phagocytic system. Most of the cases are rare complications of common infectious and neoplastic diseases, although there may be an underlying immune disorder predisposing to this syndrome. We report a case in association with immune thrombocytopenia and hemolytic anemia (Evans' syndrome). The hemophagocytic reaction appeared after a bacterial infection of the urinary tract and presented with abrupt pancytopenia and complete hemopoietic failure. We discuss the possible mechanisms of bone marrow failure related with the hemophagocytic syndrome.

16 chronic haemodialysis patients (group I), with non-microcytic anaemia (mean haemoglobin 7.2 g/dl, SD 1.0, range 5.8-9.8), moderate aluminium overload (serum aluminium 44 micrograms/l, SD 16, range 21-74), and normal or high iron stores (ferritin 800 micrograms/l SD 464, range 34-2013) were treated with intravenous desferrioxamine 1 g at the end of each dialysis for six months. 8 patients with similar characteristics served as controls (group II). After six months group I showed a rise in haemoglobin to 9.1 (SD 2.5) g/dl and a decrease in blood transfusion requirements, both significant, whereas group II showed no changes. Other significant changes observed in group I, but not group II, were a rise in reticulocytes and in red cell creatine and a fall in red cell protoporphyrin and serum ferritin. Ferritin decreased more in the patients whose anaemia improved. Minor increases in serum aluminium in group I did not differ from those in the control group. Desferrioxamine may benefit the anaemia of chronic haemodialysis patients through improvement of erythropoiesis. The effect seems not to be related to chelation of a heavy aluminium overload.

Thirty four cases of hereditary spherocytosis were studied by means of laser diffraction cytometry. The cases were grouped for study in accordance to previous splenectomy or not, familial involvement or not, and, in patients not subjected to splenectomy, severity of the clinical course. The values used to assess the presence of spherocytosis were those measuring the haemoglobin concentration within red cells, such as CH (directly estimated mean corpuscular haemoglobin), HDW (standard deviation of the distribution according to haemoglobin concentration) and % hyper (percentage of cells with haemoglobin concentration higher than 41 g/dL). The variables attained were statistically analysed by means of non-parametric tests. In patients with spherocytosis, MCV, MCHC, RDW, HDW, % hyper, and CH were significantly different from the normal group. This method points to the presence of spherocytosis by means of CH, HDW and % hyper. A reduction of the limit of haemoglobin concentration used to define % hyper (41 g/dL) could improve the sensitivity of the instrument for the diagnosis of the mild forms, which is often more difficult.

Congenital pyrimidin 5'nucleotidase deficiency manifests as hemolytic anemia with basophilic stippling. In lead poisoning, anemia, basophilic stippling and inhibition of erythrocyte pyrimidin 5' deficiency are also observed. In the present work, we report two cases of hemolytic anemia secondary to congenital deficiency of pyrimidin 5' nucleotidase and another case secondary to lead poisoning. Since 1974, when pyrimidin 5' nucleotidase deficiency was isolated, is known that hemolysis is related to the accumulation of pyrimidin nucleotides within the erythrocytes that behave as metabolic inhibitors. However, the precise metabolic process whose inhibition leads to the shortening of erythrocytes half life has not been elucidated yet.