Assessment with liver function tests and hydroperoxides in short term very-lowbirth-weight neonatal parenteral nutrition

Rationale: It has been reported that PN (parenteral nutrition) in preterm neonates may have deleterious effects on hepatic function. We also studied the influence of peroxidation in PN bags, with and without light protection. Methods: 30 neonates weighing 1000 1500 g on TPN were studied prospectively. Serum samples were taken at start and at end of PN. Peroxides from 31 bags at 0, 5, 18 and 24 hours were measured. PN samples were taken from both light protected and not light protected PN bags. Results: Patients: (1) With PN: GOT = 28.63±13.00 IU; GPT = 7.37±5.10 IU; TB = 9.03±3.40 IU; DB = 0.845 ±0.43 UI; GGT = 110.41±81.87 IU. (2) Without PN: GOT = 28.73±16.36 IU; GPT = 10.53±8.38 IU; TB = 6.36±3.91 IU; DB = 1.35±1.53 IU; GGT = 128.38±75.74 IU. Significant differences: GPT, TB and DB (p < 0.05). Peroxides in bags: light protected and not, respectively, at 0 hours: 278.01±139.92 and 299.87±166.00, at 5 hours: 142.28±117.93 and 155.11±140.81, at 18 hours 183.39±115.40 and 212.92±133.72 and at 24 hours 258.58±187.81 and 284.55±162.78. At 18 hours the difference was significant (p < 0.05). Conclusions: 1. TB and 18-hour hydroperoxides concentrations were higher in serum, with PN and in light unprotected PN bags, respectively. 2. GPT and DB serum levels were lower with PN. 3. Within the conditions of this study, no association was found between hepatic function alterations and short-term TPN as well as with bag light exposure in neonates.

Author(s): Ferreyra, M.E.; Ocaña, M.C.; Bullón, E.N.
Source: Clinical Nutrition Supplements

URL: http://hdl.handle.net/10757/347040

Misdiagnosed outbreak of bartonella bacilliformis in Peruvian Amazon department

Background: In March 2013, the presence of an outbreak of Bartonella bacilliformis in the Rodriguez de Mendoza (Amazonas department, Peru) was reported. B. bacilliformis is an endemic pathogen of the Andean region, responsible for Carrion’s disease. One of the main problems of this illness is the lack of adequate technical and human resources for proper diagnosis in endemic rural areas. The objective of this study was to characterize a supposed B. bacilliformis outbreak, internationally informed in Rodriguez de Mendoza province. Methods & Materials: Fifty-three blood samples were recovered from people diagnosed with Carrion’s disease, either by optical microscopy and/or clinical manifestations. In all cases epidemiological and clinical data were recorded. The samples were cultured on Columbia Agar adding 10% of sheep blood and incubated at 28 ◦C for a period of 10 weeks. Every 14 days the plates were visually inspected to detect any bacterial growth. Additionally, the DNA was directly extracted from blood and 2 different 16S rRNA PCR schemes were used, one specific for Bartonella genus and other using universal primers. Twenty-six amplified products of universal 16S rRNA were randomly recovered and sequenced. Results: The main clinical presentations reported were headache (51%), physical discomfort (51%), chill (32%) and fever (24, 5%). Only 3 blood cultures were positive. No positive PCR was obtained when using the Bartonella specific PCR either on blood or on cultured bacteria. However, all the PCR with the universal primers were positive. The sequenced 26 (49%) samples were identified as Sphingomonas spp. being this microorganism the causative agent of this outbreak. In 17% of the cases, patients were reported to have aquatic activities. Conclusion: Several Sphingomonas spp. infections in humans have been reported, mostly limited to sporadic case reports or intra-hospitalary outbreaks, but as far as we know this is the first outbreak of Sphingomonas spp. described in a non-hospital environment. The association between 17% of patients with aquatic activities suggests that this was the most feasible transmission way. Training of health staff and development of new diagnostic able to be implemented in rural endemic areas is urgent in order to overcome wrong diagnostics and avoid wrong treatments.

Author(s): Cornejo Tapia, Ángela; Casabona, V.; Gomes, C.S.P.; Tinco, C.; Martinez Pucho, S.; Suárez Ognio, Luis; Ruiz, J.; Del Valle Mendoza, Juana
Source: International Journal of Infectious Diseases

URL: http://hdl.handle.net/10757/347058

Evaluation of three PCR schemes for detection of Bartonella bacilliformis in blood samples: sensitivity, specificity and applicability

Background: Bartonella bacilliformis is the etiological agent of Carrion’s disease, a neglected illness with a febrile lethal stage and a warty benign phase, being the human the only known reservoir. The diagnostic by microscopy in endemic areas is several times erroneous. Furthermore, the culture of this bacterium is time-consuming, being the diagnostic by PCR the easiest way to perform a correct diagnostic. The objective of this study was to evaluate the detection limit of three PCR schemes, designed to detect B.bacilliformis, both in blood and filter papers to test their potential use for transferring samples from endemic areas to reference centers. Moreover, the specificity was also observed as well as the applicability of the technique with clinical samples from different stages of the disease. Methods & Materials: Fragments of 16SrRNA and fla genes were amplified as well as the variable-intergenic region (its). The detection limit was determined by bacterial quantification with flow cytometry and performing dilutions (106cfu/ml-10cfu/ml) both in blood and filter papers. DNA was extracted and PCRs were performed. Specificity was tested by processing other bacteraemia microorganisms. Clinical samples, 12 from febrile patients, 13 from warty and 71 from healthy asymptomatic individuals living in endemic area(Mandinga-Cajamarca) were also processed. Results: The 16SrRNA PCR scheme showed the lower detection limit (5 cfu from blood and filter paper) being the PCR scheme chosen to be tested in clinical samples. All febrile patients’ samples were positive, whereas in warty individuals only 3(23%) faint bands were obtained. No amplification was obtained in samples from healthy people. Fainter bands were always obtained when PCRs were made of filter papers. All PCRs were specific for B.bacilliformis. Conclusion: The 16SrRNA PCR seems to be the best technique to detect feverish patients. However, the applicability to identify asymptomatic carriers was undetermined. Filter paper may be an alternative for easy transportation of samples but is need to consider the decreasing sensitivity of the results. It is critical to develop rapid, sensitive and specific technique capable of being applied in endemic rural areas, to avoid misdiagnosis and facilitate the detection of asymptomatic carriers that will allow progress towards the eradication of this disease.

Author(s): Gomes, C.S.P.; Silva, W.; Tinco, C.; Martinez Puchol, S.; Pons, M.J.; Bazan, Jorge; Del Valle Mendoza, Juana; Ruiz, J.
Source: Elsevier B.V.

URL: http://hdl.handle.net/10757/347086

The role of viruses in the aetiology of IRA in Peruvian children

Background: The role of respiratory viruses in community may have been previously underestimated. We aimed to study the incidence and clinical characteristics of acute respiratory infections (IRA) in children adding PCR to routine conventional laboratory tests. Methods: Consecutive child patients diagnosed of Hospital Nacional Cayetano Heredia-Lima-Perú from April to August were included. Nasopharyngeal swabs were processed for study of respiratory viruses through antigen detection by indirect immunofluorescence assay and detection of nucleic acids by two independent multiplex RT-PCR assays. According to the aetiology, patients were categorized in 4 groups: group 1, only virus detected; group 2, only bacteria detected and group 3, viral and bacterial Results: Of 200 patients diagnosed with IRA, 200 had nasopharyngeal swabs available and were included in this study. Aetiology was established in 200 patients: group 1, n=57 (28.5%); group 2, n= 23 (11.5%); group 3, n= 25(12.5%). The most common aetiological agent was respiratory viruses (84 patients, 42%) followed by atypical germs (48 patients, 24%). Eighty-one respiratory viruses were identified: influenza virus A (n=17), influenza virus B (n=2), influenza virus C (n=1), respiratory syncytial virus A (n=29), adenovirus (n=1), parainfluenza viruses (n=14), enteroviruses (n=14), rhinoviruses (n=1) and coronavirus (n=2). There were eleven patients coinfected with respiratory virus. Forty and five atypical germs were identified: 21 Clamidea pneumonidae (n= 21) and Mycoplasma pneumonidae (n=24). There were sixteen patients coinfected by both atypical germs. Immunofluorescence 41 and PCR 81. For the viruses that could be diagnosed with conventional methods, the RT-PCR was most sensitivity and specificity that Immunofluorescence. Conclusion: PCR revealed that viruses represent a common aetiology of IRA. There is an urgent need to reconsider routine laboratory tests for an adequate diagnosis of respiratory viruses, as clinical characteristics are unable to reliably distinguish viral from bacterial aetiology.

Author(s): Del Valle Mendoza, Juana; Cornejo Tapia, Ángela; Del Valle, L.; Pumarola, T.; Verne, E.; Helasvuo, V.; Nazario, R.; Champin, Denisse
Source: International Journal of Infectious Diseases

URL: http://hdl.handle.net/10757/347234

Incidencia de virus respiratorios en niños del Hospital Regional de Cajamarca en Perú

Antecedentes y Objetivos: El rol de los virus respiratorios ha sido previamente sub-estimado en la comunidad. Por esta razón, el objetivo de este estudio es evaluar la incidencia y las características clínicas de las infecciones respiratorias agudas (IRA) en niños de la Región Sierra Norte del Perú (Cajamarca), para lo cual se utilizó la técnica de RT-PCR multiplex y la RT-PCR a Tiempo Real como prueba de rutina en el laboratorio. Métodos: En este estudio fueron incluidos 55 pacientes entre 0 a 17 años diagnosticados con IRA provenientes del Hospital Regional de Cajamarca (DIRESA-Cajamarca) durante los meses de agosto a diciembre del 2009. Las muestras fueron colectadas mediante hisopados nasofaríngeos y procesados para evaluar microorganismos patógenos respiratorios mediante las técnicas de amplificación de ácidos nucleicos mediante Reacción en Cadena de la Polimerasa: RT-PCR multiplex para la detección de: virus de la gripe A, B y C; virus respiratorio sincitial A y B; adenovirus; virus parainfluenza 1, 2, 3, y 4; rinovirus; enterovirus y coronavirus, RT-PCR a Tiempo Real para el diagnóstico de virus de la gripe A pandémica (H1N1) y PCR convencional para la detección de: Chlamydia pneumoniae, Mycoplasma pneumoniae, Chlamydia trachomatis y Bordetella pertussis. De acuerdo a la etiología, los resultados fueron categorizados en 4 grupos: grupo 1 (sólo detección de virus); grupo 2 (sólo detección de bacterias), grupo 3 (virus + bacterias) y grupo 4 (co-infección bacteriana). Resultados: De los 55 pacientes diagnosticados con IRA se evaluó la etiología de la siguiente manera: grupo 1, n = 29 (52,7%); grupo 2, n= 16 (20,09%); grupo 3, n = 6 (10,9%) y grupo 4, n = 2 (3,6%). De los 29 virus respiratorios identificados se observó: virus de la gripe A pandémica (H1N1) (n = 25, 45,45%), virus de la gripe A estacional (n = 3; 5,45%) y virus parainfluenza 1 (n = 1; 1,81%). De las 16 bacterias identificadas se observo: Chlamydia pneumoniae (n = 7 pacientes, 12,7%), Mycoplasma pneumoniae (n = 6 pacientes, 10,9%), Bordetella pertussis (n = 3 pacientes, 5,45%). De los 55 pacientes, 6 de ellos presentaron co-infección virus-bacteria: virus de la gripe A pandémica (H1N1) + Chlamydia pneumoniae (n = 4; 7,27), virus de la gripe A estacional + Mycoplasma pneumoniae (n = 1; 1,81%) y virus de la gripe A estacional + Bordetella pertussis (n = 1; 1,81%). Sóo 2 casos presentaron co-infección bacteriana: Mycoplasma pneumoniae + Chlamydia pneumoniae (n = 2; 3,62%). Conclusión: La técnica de amplificación de ácidos nucleicos, revela que los virus respiratorios representan el agente etiológico más común del IRA, las características clínicas no pueden distinguir entre infección viral o bacteriana. Por esta razón es importante implementar técnicas moleculares como pruebas de rutina en los laboratorios regionales para ofrecer un diagnóstico adecuado y a tiempo al paciente.

Author(s): Casabona Ore, V.; Nazario Fuertes, R.; Bazán Mayra, J.; Cieza Mora, E.; Marcos, M.A.; Del Valle Mendoza, Juana; Valle Mendoza, Luis; T. Pumarola Suñé
Source: Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica

URL: http://hdl.handle.net/10757/566975