RESUMO

The complex life cycle of Trichinella spiralis includes the migration of newborn larvae through the bloodstream to their encystment in muscle. The parasite establishes an intimate contact with the erythrocytes of the host both during the migration of the newborn larvae and when encysting, as this parasite causes intense vascularization in the muscle cell. The goal of this work was to study the effects of various concentrations of T. spiralis muscle larvae (ML) on erythrocyte membranes. The treatment was performed by incubating human erythrocytes with equal volume of different concentrations of ML for 30 minutes, with controlled agitation (37°C). The control erythrocytes (with no contact with the larvae) were incubated in the same way with an equal volume of physiological solution. To evaluate the alterations to the erythrocytes by the action of the larvae and in the respective controls, an Erythrocyte Rheometer and a Digital Image Analysis technique were used. The results indicated that when the larval concentration was higher, the aggregation and erythrocyte membrane alterations were also higher. Also, the erythrocyte deformability index and the erythrocyte elasticity increased. The values of isolated cell coefficient varied from 0.51 in the treatment with 100 larvae/ml to 0.91 in the incubation with 1000 larvae/ml. This experiment shows that T. spiralis muscle larvae affect significantly the red blood cell aggregation and the erythrocyte viscoelastic properties.

Assuntos

RESUMO

In this study, supplementation with the probiotic Saccharomyces boulardii promoted a reduction in intensity of infection by Toxocara canis and modulates cytokines mRNA expression in experimentally infected mice. IL-12 gene transcription had 40-fold increase in S. boulardii supplemented uninfected mice and sevenfold increase in supplemented infected mice comparing with not supplemented group. Regarding IFNγ, similar results were observed, since probiotic supplementation induced approximately 43-fold increase, but only in uninfected mice (P < 0·05). T. canis infection upregulated IL-10 expression while S. boulardii downregulated it and no change was observed for IL-4. Thus, based in these findings; we suggest that one possible mechanism responsible for S. boulardii protection effect against T. canis infection is by the modulation of cytokines expression, especially IL-12.

Assuntos

RESUMO

This study provides direct and indirect evidence of temporally and spatially consistent spawning aggregations for the grouper Mycteroperca olfax. Recently reported declines in population numbers, probably related to the direct targeting of aggregations by artisanal fishermen, highlight the urgent need for species-specific management actions in the Galapagos Marine Reserve, such as minimum and maximum landing sizes, and the importance of protecting key aggregation sites with the declaration of no-take areas and the establishment of total fishing bans during the reproductive season.

Assuntos

RESUMO

The objective was to evaluate the effect of three cryopreservation methods on the in vitro maturation (IVM) and membrane integrity (MIn) of immature equine oocytes. An open pulled straw (OPS) method, a novel solid surface vitrification (SSV) process, and the addition of a synthetic ice blocker were evaluated. Compared with the control group (N=269), the OPS (N=159) and the SSV (N=202) cryopreservation methods decreased both IVM (50.9 vs. 13.3 and 9.4%, respectively; P<0.001) and MIn (76.6 vs. 31.1 and 33.7%; P<0.001) of immature equine oocytes. However, inclusion of 0.1% ice blocker in the OPS vitrification process increased the rates of both IVM (30.5%; P<0.01) and MIn (45.8%; P<0.05) of the oocytes (N=59). Including 0.1% ice blocker in the SSV process improved the IVM rate (20.9%; P<0.05), whereas MIn remained compromised in this group (N=67). However, increasing the concentration of the ice blocker (to 1.0%) in the cryopreservation methods did not significantly improve rates of IVM. In conclusion, the addition of a synthetic ice blocker (0.1%) to both cryopreservation processes significantly increased rates of both IVM and MIn of immature equine oocytes cryopreserved by OPS.

Assuntos

RESUMO

The objective was to introduce exogenous DNA into commercially sex-sorted bovine sperm using nanopolymer for transfection. In the first experiment, the optimal concentration and ratio of linear-to-circular plasmid was determined for NanoSMGT in unsorted sperm. A second experiment was conducted to transfect exogenous DNA into sex-sorted sperm. Exogenous DNA uptake occurred in a dose-dependent manner (P < 0.05). The optimal amount of DNA was 10 µg/10(6) cells. The ratios of linear-to-circular plasmid do not influence the uptake by unsorted sperm cells and none of the tested treatments affected sperm motility and viability. Commercially sex-sorted bovine sperm were able to uptake exogenous DNA using nanopolymer; however, both X- and Y-sorted sperm had decreased DNA uptake in comparison to unsorted sperm (P < 0.05). Neither sperm motility nor viability were affected by nanotransfection. In conclusion, nanopolymer efficiently introduced exogenous DNA into commercially sex-sorted bovine sperm; we inferred that these sperm could be used for production of embryos of the desired sex, a technique named NanoSMGT.

Assuntos

Assuntos

Assuntos

Assuntos

RESUMO

A new species of Cercocarpus, Cercocarpus mixteca Velasco de León & Cevallos-Ferriz, is described based on leaf impressions from the Los Ahuehuetes locality, near Tepexi de Rodríguez, Puebla, Mexico. The lamina is obovate, 1.3cm in length by 0.5cm in width, has a serrate margin in its distal fourth, craspedodromous venation with a single straight mid-vein and two to four pairs of secondary ones, and areols that tend to be quadrangular in shape. A phenetic analysis of the agglomerative, non-hierarchical type, with mean linkage, is applied using 22 OTUs and 34 character states. The morphological characters observed on the leaves of the new fossil plants support the recognition of a new taxon closely related to the extant Cercocarpus paucidentatus growing naturally in northern Mexico. Its microphyll size corresponds with the temperate to xeric climate postulated for the Los Ahuehuetes locality; this further suggests that some taxa, like Cercocarpus, have a long history in low latitude North America. In this particular case, the extant Cercocarpus fothergilloides and Cercocarpus macrophyllus could, as they were able to colonise new humid and xeric areas, represent descendants of C. mixteca.

RESUMO

Genomic fingerprints from 92 capsulated and noncapsulated strains of Haemophilus influenzae from Mexican children with different diseases and healthy carriers were generated by PCR using the enterobacterial repetitive intergenic consensus (ERIC) sequences. A cluster analysis by the unweighted pair-group method with arithmetic averages based on the overall similarity as estimated from the characteristics of the genomic fingerprints, was conducted to group the strains. A total of 69 fingerprint patterns were detected in the H. influenzae strains. Isolates from patients with different diseases were represented by a variety of patterns, which clustered into two major groups. Of the 37 strains isolated from cases of meningitis, 24 shared patterns and were clustered into five groups within a similarity level of 1.0. One fragment of 1.25 kb was common to all meningitis strains. H. influenzae strains from healthy carriers presented fingerprint patterns different from those found in strains from sick children. Isolates from healthy individuals were more variable and were distributed differently from those from patients. The results show that ERIC-PCR provides a powerful tool for the determination of the distinctive pathogenicity potentials of H. influenzae strains and encourage its use for molecular epidemiology investigations.

Assuntos

RESUMO

Cryptosporidium parvum, Isospora belli, Cyclospora cayetanensis and Microsporidia are frequent pathogens in the immunodeficient host, which may cause multiple infections. The above mentioned parasites are found in feces by the application of different specific tintorial techniques. The objective of this work was the development of a stain for the simultaneous detection of these parasites, reducing costs as well as the time taken to make the diagnosis. The safranin-trichrome stain is simple, chip and its results are similar to those of specific tints. All microorganisms are easy to detect and besides being perfectly distinguishable from fungi and faecal elements.

Assuntos

RESUMO

We worked with 185 middle-class patients above 18 years of age, both sexes, who presented diarrhea and/or chronic gastrointestinal disorders. The faeces were collected serially in formol 10% and processed in the following way: direct microscopy, with and without wet staining, concentration by Ritchie's method, 1% safranine technique for a specific investigation of Cryptosporidium sp., and faecal sieving macroparasites. Twenty eight point six of the studied patients showed at least one enteroparasite in their faeces, 48 harboured one parasite and 5 harboured two parasites. The following parasites were found and their corresponding percentages in the entire studied population are given below: Blastocystis hominis 15.7%, Giardia lamblia 7.5%, Cryptosporidium sp. 1.6%, Entamoeba coli 3.3%, Chilomastix mesnilii 1.1%, Ancylostoma duodenale-Necator americanus 0.5%, Ascaris lumbricoides 0.5%, Enterobious vermicularis 0.5% y Endolimax nana 0.5%. The most frequently found enteroparasites in the positive patients were B. hominis and G. lamblia. Cryptosporidium sp. was diagnosed in only three patients. The source of infection could be presumed in all of them. The symptomatology coincided with that described for this coccid in the bibliography. In spite of the fact that they were HIV seronegative patients the diarrhea was not self-limiting, but the immunologic profile of their relatives remained unknown and no other cause of immunosuppression could be detected with justified chronicity. The treatment with spiramycin was effective. Giardiasis was found in 17 patients, and the source of infection could not be inferred in any of them. They all had chronic diarrhea and their most frequent symptoms were abdominal pain, metallic taste, flatulency and nausea. Most of these patients were harboured one parasite, and only 2 of them simultaneously presented another faecal parasite associated to G. lamblia. Treatment with metronidazole was successful in all of them. Twenty nine patients were found to have B. hominis. The source of infection could not be inferred, this amoeboid was present as the only parasite in 25 patients. Predominant symptoms were flatulence, abdominal distention and colis. All patients suffered from chronic diarrhea, alternating, in some cases, with constipation. Good therapeutic results were obtained with metronidazole. Considering that one third of the patients examined presented faecal parasites associated to chronic disorders, it is important to insist on the detection of parasites to chronic disorders, it is important to insist on the detection of parasites using appropriate diagnostic techniques since the application of specific therapy made their eradication possible as well as relieving the patients' symptomatology.

Assuntos

RESUMO

For 41 months, the presence of parasites was investigated in a seropositive HIV population with the clinical characteristics of stages 3 and 4 according to the OMS classification; of 212 fecal samples belonging to 135 patients which were analyzed, 53.33% presented enteroparasites. A direct parasitological exam and a Ritchie concentration were performed on the feces collected in formol 10%. Two smears were stained with Safranine 1% and two with modified Ziehl-Nielsen to identify Cryptosporidium sp. The detected frequencies were: Cryptosporidium The detected frequencies were: Cryptosporidium sp. 11.11%; I. belli 2.96%; G. lamblia 11.85%; B. hominis 26.66%; A. lumbricoides 2.96%; E. vermicularis 1.48%; H. nana 0.74%; E. coli 13.33%; E. nana 5.93%; Ch. mesnilii 2.22% and I. butschlii 0.74%. There were 46 monoparasitized patients, 19 biparasitized, 5 triparasitized and 2 tetraparasitized. Furthermore, 17 bronchoalveolar lavages (BAL) and 194 sputa were processed, collected in formol 10% and centrifuged to exhaustion; 10 smears were prepared with sediment and were stained with toluidine blue. Groccot (Gomori) coloration was used to confirm doubtful cases. In 47% of the BAL and in 22,68% of the sputa P. carinii was diagnosed. This represents 34.68%. The percentage of positive cases was: 30.88% for those patients who sent a single sputum, 36.84% for those who sent more than one and 27.27% for BAL. Finally, in 7 patients who sent BAL and sputa, there were 2 positive and 2 negative cases in both materials, while P. carinii was diagnosed in 3 patients only in their BAL.

Assuntos

RESUMO

For 41 months, the presence of parasites was investigated in a seropositive HIV population with the clinical characteristics of stages 3 and 4 according to the OMS classification; of 212 fecal samples belonging to 135 patients which were analyzed, 53.33

presented enteroparasites. A direct parasitological exam and a Ritchie concentration were performed on the feces collected in formol 10

. Two smears were stained with Safranine 1

and two with modified Ziehl-Nielsen to identify Cryptosporidium sp. The detected frequencies were: Cryptosporidium The detected frequencies were: Cryptosporidium sp. 11.11

; I. belli 2.96

; G. lamblia 11.85

; B. hominis 26.66

; A. lumbricoides 2.96

; E. vermicularis 1.48

; H. nana 0.74

; E. coli 13.33

; E. nana 5.93

; Ch. mesnilii 2.22

and I. butschlii 0.74

. There were 46 monoparasitized patients, 19 biparasitized, 5 triparasitized and 2 tetraparasitized. Furthermore, 17 bronchoalveolar lavages (BAL) and 194 sputa were processed, collected in formol 10

and centrifuged to exhaustion; 10 smears were prepared with sediment and were stained with toluidine blue. Groccot (Gomori) coloration was used to confirm doubtful cases. In 47

of the BAL and in 22,68

of the sputa P. carinii was diagnosed. This represents 34.68

. The percentage of positive cases was: 30.88

for those patients who sent a single sputum, 36.84

for those who sent more than one and 27.27

for BAL. Finally, in 7 patients who sent BAL and sputa, there were 2 positive and 2 negative cases in both materials, while P. carinii was diagnosed in 3 patients only in their BAL.

RESUMO

We worked with 51 samples, 7 bronchoalveolar lavages (BAL) and 44 sputa (S) of 31 AIDS patients with clinical and radiographic symptoms compatible with Pneumocystis pneumonia. With the aim of finding a specific sensitive methodology for the diagnosis of Pneumocystis carinii, we evaluated 4 coloration techniques (silver methenamine, its modification without gold chloride, toluidine blue and Giemsa). 35% of the patients studied were positive. P. carinii were observed in 18% of the 44 sputa. We observed that the analysis of a single sputum sample (S) has a very low sensitivity and that the processing of two or more samples is necessary since only one of the 14 patients who had sent a single sample was found P. carinii positive, while in the remaining ten who had sent more than one (S) sample, the microorganism was detected in 50%. 4 of the 7 BAL were positive. 4 BAL were preceded by the analysis of an (S) sample: in two cases the results were negative while BAL allowed us to make the diagnosis, thus demonstrating its greater efficacy. To enhance sensitivity each sample was centrifuged until exhaustion and 10 slides were prepared for coloration with the final sediment. The four techniques employed were specific and all the Pneumocystis pneumonia patients responded to the treatment. Silver methenamine, its modification without gold chloride, and toluidine blue were very sensitive, in contrast to of Giemsa. The stain to be chosen is either silver methenamine, or its modification, because both achieve the best contrast, allowing optimum P. carinii identification. We suggest the implementation of some of these techniques in laboratory routine.

Assuntos

RESUMO

We worked with 51 samples, 7 bronchoalveolar lavages (BAL) and 44 sputa (S) of 31 AIDS patients with clinical and radiographic symptoms compatible with Pneumocystis pneumonia. With the aim of finding a specific sensitive methodology for the diagnosis of Pneumocystis carinii, we evaluated 4 coloration techniques (silver methenamine, its modification without gold chloride, toluidine blue and Giemsa). 35

of the patients studied were positive. P. carinii were observed in 18

of the 44 sputa. We observed that the analysis of a single sputum sample (S) has a very low sensitivity and that the processing of two or more samples is necessary since only one of the 14 patients who had sent a single sample was found P. carinii positive, while in the remaining ten who had sent more than one (S) sample, the microorganism was detected in 50

. 4 of the 7 BAL were positive. 4 BAL were preceded by the analysis of an (S) sample: in two cases the results were negative while BAL allowed us to make the diagnosis, thus demonstrating its greater efficacy. To enhance sensitivity each sample was centrifuged until exhaustion and 10 slides were prepared for coloration with the final sediment. The four techniques employed were specific and all the Pneumocystis pneumonia patients responded to the treatment. Silver methenamine, its modification without gold chloride, and toluidine blue were very sensitive, in contrast to of Giemsa. The stain to be chosen is either silver methenamine, or its modification, because both achieve the best contrast, allowing optimum P. carinii identification. We suggest the implementation of some of these techniques in laboratory routine.

RESUMO

We worked with 51 samples, 7 bronchoalveolar lavages (BAL) and 44 sputa (S) of 31 AIDS patients with clinical and radiographic symptoms compatible with Pneumocystis pneumonia. With the aim of finding a specific sensitive methodology for the diagnosis of Pneumocystis carinii, we evaluated 4 coloration techniques (silver methenamine, its modification without gold chloride, toluidine blue and Giemsa). 35

of the patients studied were positive. P. carinii were observed in 18

of the 44 sputa. We observed that the analysis of a single sputum sample (S) has a very low sensitivity and that the processing of two or more samples is necessary since only one of the 14 patients who had sent a single sample was found P. carinii positive, while in the remaining ten who had sent more than one (S) sample, the microorganism was detected in 50

. 4 of the 7 BAL were positive. 4 BAL were preceded by the analysis of an (S) sample: in two cases the results were negative while BAL allowed us to make the diagnosis, thus demonstrating its greater efficacy. To enhance sensitivity each sample was centrifuged until exhaustion and 10 slides were prepared for coloration with the final sediment. The four techniques employed were specific and all the Pneumocystis pneumonia patients responded to the treatment. Silver methenamine, its modification without gold chloride, and toluidine blue were very sensitive, in contrast to of Giemsa. The stain to be chosen is either silver methenamine, or its modification, because both achieve the best contrast, allowing optimum P. carinii identification. We suggest the implementation of some of these techniques in laboratory routine.

RESUMO

We worked with 51 samples, 7 bronchoalveolar lavages (BAL) and 44 sputa (S) of 31 AIDS patients with clinical and radiographic symptoms compatible with Pneumocystis pneumonia. With the aim of finding a specific sensitive methodology for the diagnosis of Pneumocystis carinii, we evaluated 4 coloration techniques (silver methenamine, its modification without gold chloride, toluidine blue and Giemsa). 35

of the patients studied were positive. P. carinii were observed in 18

of the 44 sputa. We observed that the analysis of a single sputum sample (S) has a very low sensitivity and that the processing of two or more samples is necessary since only one of the 14 patients who had sent a single sample was found P. carinii positive, while in the remaining ten who had sent more than one (S) sample, the microorganism was detected in 50

. 4 of the 7 BAL were positive. 4 BAL were preceded by the analysis of an (S) sample: in two cases the results were negative while BAL allowed us to make the diagnosis, thus demonstrating its greater efficacy. To enhance sensitivity each sample was centrifuged until exhaustion and 10 slides were prepared for coloration with the final sediment. The four techniques employed were specific and all the Pneumocystis pneumonia patients responded to the treatment. Silver methenamine, its modification without gold chloride, and toluidine blue were very sensitive, in contrast to of Giemsa. The stain to be chosen is either silver methenamine, or its modification, because both achieve the best contrast, allowing optimum P. carinii identification. We suggest the implementation of some of these techniques in laboratory routine.

RESUMO

We worked with 51 samples, 7 bronchoalveolar lavages (BAL) and 44 sputa (S) of 31 AIDS patients with clinical and radiographic symptoms compatible with Pneumocystis pneumonia. With the aim of finding a specific sensitive methodology for the diagnosis of Pneumocystis carinii, we evaluated 4 coloration techniques (silver methenamine, its modification without gold chloride, toluidine blue and Giemsa). 35

of the patients studied were positive. P. carinii were observed in 18

of the 44 sputa. We observed that the analysis of a single sputum sample (S) has a very low sensitivity and that the processing of two or more samples is necessary since only one of the 14 patients who had sent a single sample was found P. carinii positive, while in the remaining ten who had sent more than one (S) sample, the microorganism was detected in 50

. 4 of the 7 BAL were positive. 4 BAL were preceded by the analysis of an (S) sample: in two cases the results were negative while BAL allowed us to make the diagnosis, thus demonstrating its greater efficacy. To enhance sensitivity each sample was centrifuged until exhaustion and 10 slides were prepared for coloration with the final sediment. The four techniques employed were specific and all the Pneumocystis pneumonia patients responded to the treatment. Silver methenamine, its modification without gold chloride, and toluidine blue were very sensitive, in contrast to of Giemsa. The stain to be chosen is either silver methenamine, or its modification, because both achieve the best contrast, allowing optimum P. carinii identification. We suggest the implementation of some of these techniques in laboratory routine.