An extensive field study reveals the circulation of new genetic variants of subtype 1a of bovine viral diarrhea virus in Uruguay.

Bovine viral diarrhea virus (BVDV) is a major pathogen worldwide, causing significant economic losses to the livestock sector. In Uruguay, BVDV seroprevalence at the farm level is >80%. In this work, 2546 serum, blood or tissue samples collected from animals suspected of being affected by BVD between 2015 and 2017 were analyzed by reverse transcription PCR and sequencing. Analysis of the BVDV genomic regions 5'UTR/Npro, Npro and E2 revealed that BVDV-1a, 1i and 2b circulate in the country, with BVDV-1a being the most prevalent subtype. Population dynamics studies revealed that BVDV-1a has been circulating in our herds since ~1990. This subtype began to spread and evolve, accumulating point mutations at a rate of 3.48 × 10-3 substitutions/site/year, acquiring specific genetic characteristics that gave rise to two local genetic lineages of BVDV-1a. These lineages are divergent from those circulating worldwide, as well as the vaccine strain currently used in Uruguay. The most notable differences between field and vaccine strains were found in the E2 glycoprotein, suggesting that the amino acid substitutions could result in failure of cross-protection/neutralization after vaccination. This is the first study that compares Uruguayan BVDV field and vaccine strains with other BVDV strains from throughout the world. The results obtained in this study will be very useful for developing a suitable immunization program for BVDV in Uruguay by identifying local field strains as candidates for vaccine development.

Molecular diversity of bovine viral diarrhea virus in uruguay.

Bovine viral diarrhea (BVD) affects bovine production and reproduction causing significant economic losses all over the world. Two viral species has been recognized: BVDV-1 and BVDV-2, both distributed worldwide. Recently, novel specie of BVDV named HoBi-like pestivirus was discovered. The presence of BVDV was confirmed in 1996 in Uruguay, however, does not exist until today a schedule of compulsory vaccination along the country. Serological studies with samples from all Uruguayan herds were performed during 2000 and 2001 demonstrating that all of them were seropositive to BVDV with a mean prevalence of 69%. In addition, there have been no new studies done since those previously described and it is important to mention that the genetic diversity of BVD has never been described in Uruguay. Nowadays, there is strongly suspect that BVDV is one of the most important causes of reproductive failures in our herds. The aim of this study was to describe for the first time in Uruguay the genetic diversity of BVDV with samples collected from different regions along the country. Serological status of 390 non-vaccinated animals against BVDV with reproductive problems from farms of Rivera, Tacuarembó and Florida departments of Uruguay were studied. All herds were seropositive to BVDV and high proportion of animals were positive (298/390), while 4.1% (16/390) of the animals were positive to Antigen Capture ELISA test and Real Time PCR. Phylogenetic analysis performed with concatenated sequences from the 5'UTR and Npro genomic regions revealed that BVDV-1 and BVDV-2 are infecting our herds, being BVDV-1 the most frequently found. The major subtype was BVDV-1a, followed by BVDV-1i and BVDV-2b. This is the first study that describes the genetic diversity of BVDV in Uruguay and it will contribute to the elaboration of sanitization programs.

Isolation and characterization of canine parvovirus type 2C (CPV-2C) from symptomatic puppies.

Canine parvovirus type 2 (CPV-2) is a leading cause of diarrhea in puppies in several parts of the world. In this study CPV-2 was detected and recovered from puppies showing clinical disease from Montevideo, Uruguay. Samples were processed and used to infect CRFK and MDCK cells in order to isolate the virus. Out of twelve, two samples were positive for CPV-2. A genomic region of 583 bp was amplified and the molecular characterization was performed by sequencing, phylogenetic analysis and Restriction Fragment Length Polymorphism (RFLP). Two isolated viruses (UY1 and UY2) were CPV-2c-like viruses. The comparison between the cytophatic effect (CPE) of CPV-2 (vaccinal virus) and CPV-2c (isolated virus) on primary canine cells cultures and on CRFK line cells, demonstrated that CPV-2c is less citopathogenic in CRFK than in primary cultures. Our study represents the first report on isolation and characterization of canine parvovirus type 2c (CPV-2c) in cell cultures from South American dogs.

Phylogenetic comparison of the carboxy-terminal region of glycoprotein C (gC) of bovine herpesviruses (BoHV) 1.1, 1.2 and 5 from South America (SA).

Different types and subtypes of bovine herpesvirus 1 and 5 (BoHV-1 and BoHV-5) have been associated to different clinical conditions of cattle, in such a way that type/subtype differentiation has become an essential tool for understanding the pathogenesis and epidemiology of BoHV infections. In search for a genomic region that would allow a clear distinction between BoHV-1 and BoHV-5, the carboxy-terminal portion of glycoprotein C (gC), corresponding to residues 321-450 (BoHV-1) and 301-429 (BoHV-5) of 23 South American (SA) isolates (Brazil mostly) was amplified and sequenced. The nucleotide sequence alignments revealed levels of genomic similarity ranging from 98.7 to 99.8% among BoHV-1 isolates, 88.3 to 92% between BoHV-1/BoHV-5 and 96 to 99.7% among BoHV-5 isolates. At the amino acid level, sequence similarity varied ranging from 97.5 to 99.5% among BoHV-1, 77.5 to 84.4% between BoHV-1/BoHV-5 and 92.1 to 99.5% (BoHV-5/BoHV-5). The isolates could be clearly separated into BoHV-1.1, BoHV-1.2 and BoHV-5 after phylogenetic analysis. The results suggest that the phylogenetic analysis performed here can be used as a potential molecular epidemiological tool for herpesviruses.

Increased spinal c-Fos expression with noxious and non-noxious peripheral stimulation after severe spinal contusion.

The effects of severe contusive spinal cord injury (SCI), at thoracic level 8 (T8), on lumbar c-Fos expression in the spinal cord was investigated. As hypothesized, chronic SCI has a significant effect on expression of c-Fos in the dorsal spinal sensory areas with noxious and innocuous peripheral stimulation of the sciatic nerve. This alteration to stimulation effects was measured using counts of c-Fos immunoreactive cells in the dorsal horn of the L5 lumbar spinal cord in injured animals at 90 days post-injury and in uninjured controls. The number of c-Fos immunoreactive cells increased in SCI rats only after noxious peripheral stimulation (electrical and chemical) suggesting a general increase in excitability in spinal pathways (central sensitization) associated with chronic SCI. These altered responses may represent a functional anatomical reorganization of spinal cord circuitry leading to increased dorsal horn c-Fos expression as a response to severe chronic contusive damage to the spinal cord sensory pathways.

Alteration in diastolic function following cardiac cryopreservation at subzero temperatures.

BACKGROUND: We have studied the alterations produced in the diastolic function of the left ventricle after applying a protocol of cryopreservation at subzero temperatures. METHODS: Isolated rabbit hearts and 5% polyethylene glycol (PM 4000) as the cryoprotective agent were used for the study. RESULTS-CONCLUSIONS: Following cryopreservation we found a statistically significant increase in systolic function. However, the diastolic function shows worsening, with a statistically significant increase (p < 0.05) in mean stiffness, decrease in differential stiffness, (p < 0.05) and upward and leftward displacement of the diastolic pressure-volume curve.

[Cardiac cryopreservation at subzero temperatures: study of systolic and diastolic function]. / Criopreservación cardíaca a temperatura subcero: estudio de la función sistólica y diastólica.

UNLABELLED: We studied the alterations produced in left ventricular systolic and diastolic function after applying a protocol of cryopreservation at subzero temperatures. Isolated rabbit hearts were used for the study with 5% polyethylene glycol (PM 4000) being the cryoprotective agent. MATERIALS AND METHODS: The cryoprotectant solution CP-16 was used on the explanted heart in three phases: induction, storage and thawing. After 60 minutes at -1.6 C and thawing at 2.7 C/min, the heart was connected to a Langendorff system and perfused anterogradely with Krebs-Henseleit buffer. We analyzed the systolic and diastolic parameters before and after cryopreservation, thereby establishing a comparative statistical study. RESULTS: Following cryopreservation we found a statistically significant increase (p < 0.05) in the peak and developed pressure of the left ventricle with an upward, left displacement of the ventricular function curve. This is indicative of improvement in systolic function. However, the diastolic function showed worsening, with a statistically significant increase (p < 0.05) in mean stiffness, decrease in differential stiffness (p < 0.05) and upward, left displacement of the diastolic pressure-volume curve. CONCLUSIONS: On the basis of our results we concluded that: a) PM 4000 polyethylene glycol maintains the heart biological viability during cryopreservation at subzero temperatures, and b) after an cryopreservation left ventricular diastolic function worsens with an increase in systolic function.

Isolation and characterization of canine parvovirus type 2c (CPV-2c) from symptomatic puppies

Canine parvovirus type 2 (CPV-2) is a leading cause of diarrhea in puppies in several parts of the world. In this study CPV-2 was detected and recovered from puppies showing clinical disease from Montevideo, Uruguay. Samples were processed and used to infect CRFK and MDCK cells in order to isolate the virus. Out of twelve, two samples were positive for CPV-2. A genomic region of 583 bp was amplified and the molecular characterization was performed by sequencing, phylogenetic analysis and Restriction Fragment Length Polymorphism (RFLP). Two isolated viruses (UY1 and UY2) were CPV-2c-like viruses. The comparison between the cytophatic effect (CPE) of CPV-2 (vaccinal virus) and CPV-2c (isolated virus) on primary canine cells cultures and on CRFK line cells, demonstrated that CPV-2c is less citopathogenic in CRFK than in primary cultures. Our study represents the first report on isolation and characterization of canine parvovirus type 2c (CPV-2c) in cell cultures from South American dogs.

Disección espontánea de arteria ilíaca asociada a ejercicio y uso de ergotamina / Spontaneous dissection of iliac artery related with excercise and ergotamine. A case report

Introduction: Spontaneous dissection of the iliac artery (SDIA) is an extremely rare clinical manifestation, associated to different etiologies, and it usually shows an ischemia of the involved lower extremity. Clinical case: We report a case of a 48-year-old man, with past medical history of chronicle consumption of ergotamine, which presented left lower limb ischemia, while practicing physical exercise. An emergent contrast-enhanced computed tomography scan showed a spontaneous dissection of the common and the external left iliac artery. An endovascular therapy of the lesion was performed with self-expanding stents, achieving the reconstruction of the lesion, the recovery of the blood flow and of the lower limb ischemia. Conclusions: There are no previous descriptions of the association between ergotamine consumption, sport and this very rare pathology. Endovascular treatment represents a less invasive and, such as in our case report, successful management of the SDIA, and it should be considered among the alternative therapies.

Introducción: La disección espontánea de la arteria ilíaca es un cuadro muy poco frecuente asociado a diferentes etiologías que habitualmente se presenta como isquemia de la extremidad comprometida. Caso clínico: Se reporta el caso de un paciente masculino de 48 años con antecedente de consumo crónico de ergotamina, quien mientras practicaba deporte presenta cuadro de isquemia aguda de la extremidad inferior izquierda. Angio tomografía computada demostró disección espontánea de la arteria ilíaca común y externa izquierda. Se realizó terapia endovascular de la lesión con stents auto expandibles, logrando la reparación de la lesión, el restablecimiento del flujo y la recuperación de la isquemia de la extremidad. Discusión y conclusiones: No existen reportes previos de esta patología poco frecuente, en que se asocie en forma conjunta la práctica de deporte y el uso de ergotamina. Dentro de las alternativas terapéuticas, la reparación endovascular representa una opción menos invasiva y, como en este caso, con óptimos resultados.

[One year follow up of successful coronary angioplasty in non selected patients]. / Seguimiento clínico al año de angioplastias coronarias exitosas en pacientes no seleccionados.

BACKGROUND: Re-stenosis after percutaneous coronary angioplasty (PTCA) is related to clinical and angiographic features. AIM: To describe the clinical and angiographic characteristic of our patients with coronary cardiopathy subjected to PTCA and the predictor factors for re-stenosis. MATERIAL AND METHODS: We gathered the clinical and angiographic characteristics of all patients who underwent a successful PTCA of a native coronary artery. All patients had a clinical assessment one year after the procedure. Patients were classified in Group 1, if they did not have angina or coronary events after the angioplasty or Group 2, if they had angina or a coronary event after the procedure. Only Group 2 patients were subjected to a coronary angiogram. RESULTS: We collected 383 PTCA procedures. Follow up information was obtained in 92.2%. Three hundred forty two patients (89.3%) were assessed one year the procedure. Nine patients (2.3%) died of a cardiovascular cause. Ninety patients (26.3%) were classified in Group 2. In 65 patients, angiographic re-stenosis was demonstrated (19%). Re-stenosis occurred in 36 and 13% of patients with an without Diabetes Mellitus, respectively (p <0.01). The other clinical predictor variables were a history of myocardial infarction (p =0.007), obesity (p =0.041) and hypercholesterolemia (p =0.050). None of the angiographic characteristics predicted restenosis. Stents were protective factors against restenosis (15.6% in stented lesions vs 25.4% in nonstented; p =0.01). CONCLUSIONS: Re-stenosis after angioplasty occurred in 19% of our patients with angina or coronary events. The clinical variables associated with a higher risk of re-stenosis were diabetes (the main risk factor), previous myocardial infarction, obesity and hypercholesterolemia. Angiographic variables were not associated with re-stenosis. The use of stents decreases the incidence of re-stenosis in all groups).

[Maternal perception of the quality of care given to their children during hospitalization]. / Percepción materna de la calidad en la atención a niños hospitalizados.

In order to evaluate the perception of mothers on the care given to their hospitalized children, a survey was specially designed and carried out with 100 mothers on the day of their child's discharge. Hospital care was rated excellent or good by a 96%, regular or bad by 4% of surveyed mothers. Most (74%) mothers saw at least one member of the caring team acting affectionately with their children; only 2% complained that her child was reprimanded or punished. Lack of equipment or human resources and comfort facilities were frequently perceived by mothers but quality of care was generally considered to be satisfactory. This kind of studies is probably useful to improve community's health services.

[Growth, acceptance, and tolerance with a new milk formula]. / Crecimiento, aceptación y tolerancia con una nueva fórmula láctea.

The purpose of this investigation was to evaluate the effect of a modified cow's milk formula on growth, tolerance and acceptability, which was studied in 2,097 infants selected from those controlled in primary health centers at Santiago, Chile: 1,119 were fed a modified cow's milk formula (LPM), while an unmodified powdered cow's milk (LP) was given to 978 controls of the same age and conditions, as well as solid foods after 4 months of age. Both experimental and control groups were followed along a 4 months period for records of anthropometric data, morbidity, acceptability and tolerance for LP and LPM. We did not find significant differences in growth (values ranging 95 to 100% of NCHS standards W/A or H/A). A slightly higher energy intake and lower protein intake was observed with LPM. Gastrointestinal symptoms were infrequently seen and of similar frequency and character in both 0 to 4 month old groups (4.3% with LPM vs. 5.3% with LP at first control). These gastrointestinal signs were more frequently recorded under LPM than with LP in the 8 to 12 months old group only at first control (colics 8% vs. 4% p < 0.01 and abnormal stools 9.4% vs. 5.5% p < 0.01). Acute acceptability expressed as percentage withdrawal from study was not significantly lower with LPM than LP (5 to 15% vs. 2% N.S.). The proposed modified formula seems to be a good alternative to powdered cow's milk for infants.

[Virologic diagnosis of an outbreak of epidemic hemorrhagic conjunctivitis by an indirect immunofluorescence technique. Cuba.1989]. / Diagnóstico virológico de un brote de conjuntivitis hemorrágica epidémica mediante la técnica de inmunofluorescencia indirecta. Cuba, 1989.

The causative agent of an hemorrhagic conjunctivitis outbreak was assessed through the study of 18 conjunctival exudates using the indirect immunofluorescence technique and isolation in cell culture of human embryo fibroblasts. In addition, 395 paired serum samples were studied and Enterovirus 70 was found to be responsible for the outbreak.

[Evaluation of a new technology for the detection of enterovirus in sewage]. / Valoración de una novedosa tecnología para la detección de enterovirus en aguas negras.

This paper reported the appraisal of a novel technology for the detection of enterovirus in sewage based on a viral recovery method using polyethylenglycol as a concentrating agent and on the combined use of viral isolation and polymerase chain reaction as viral detection and identification techniques. It was also confirmed that the viral recovery method is highly efficient since it allows to recover all viruses present in sewage and to preserve their infective capacity. It was proved that the polymerase chain reaction (PCR) is less sensitive than cell culture for detecting enteroviruses in sewage. It is also possible to use guadinine isothiocyanate, whose components can be prepared in the lab, as an alternative method for taking out and purifying nucleic acids instead of using the conventional TRIZOL method which is the one recommended in these cases by the World Health Organization.

Treatment of children suffering from severe malnutrition in an area of Santiago, Chile.

PIP: Despite marked progress in recent years, infant and young child malnutrition remains a serious health problem in Chile. To help define the nature of that problem better, researchers working in the Southeastern Metropolitan Area of Santiago conducted a survey of infants classified as having grade 2 protein-calorie malnutrition (PCM) and children under age 2 classified as having grade 3 PCM during the period March 31-October 1, 1980. The survey, based on a review of health facility records for all children in the area receiving care during the study period, found a total of 388 infants with grade 2 PCM and 102 subjects under 2 with grade 3 PCM. The study also found that the subjects who received comprehensive professional outpatient care at the area's Infant Nutrition Center fared significantly better than those cared for at outpatient health centers. The authors therefore recommend that the treatment of such children should be referred to or coordinated with centers that are diagnostically and therapeutically more sophisticated than the outpatient health centers. In this vein, it appears that the style of treatment provided by the Infant Nutrition Center could be developed more broadly throughout Chile and could be effectively employed in other countries with similar situations. It also appears, however, that malnourished children cannot be treated effectively as outpatients because they have very unsatisfactory home environments. One alternative approach is such cases is to admit the child as an inpatient to a special nutritional recovery center. Hospitalization, which typically occurs when the child is suffering from 1 or more pathologic conditions besides malnutrition, constitutes another alternative. However, the present survey found that the nutritional status of hospitalized study children did not improve, on the average, and some subjects experienced a marked nutritional deterioration. Another important point is that over 1/4 of the malnourished study children had low birthweights or had been premature. This underscores the importance of avoiding low birthweights as much as possible, and then giving low birthweight infants adequate care and treatment. In addition, clear standards should be established for the feeding of low birthweight children and for determining their nutritional status.^ieng

[The antigenic characteristics of the viruses isolated from the cerebrospinal fluid of patients with epidemic neuropathy]. / Características antigénicas de los virus aislados del líquido cefalorraquídeo de pacientes con neuropatía epidémica.

During the epidemic of neuropathy occurred in Cuba from 1992 to 1994 viral agents antigenically related to the Coxsackie viruses were isolated from the cerebrospinal fluid of patients. To establish a function of these viruses in the etiopathogenesis of the disease, strains 47/93 IPK identified as Coxsackie A9 and the strain 44/93 of light cytopathic effect were selected to conduct a study of their antigenic characteristics by Western Blot. It was proved that there is an antigenic relationship between both strains and it was also demonstrated the absence of structural proteins in their native form among the agents of light cytopathic effect. According to these results, it is stated the possibility that persistence may be a mechanism through which these viruses take part in the etiopathogenesis of epidemic neuropathy in Cuba.

[Rapid detection of Enterovirus by a direct method of polymerase chain reaction]. / Detección rápida de Enterovirus mediante un método directo de reacción en cadena de la polimerasa.

For the detection of Enterovirus, we devised a direct economical method of polymerase chain reaction which does not require a previous extraction of ribonucleic acid and uses infected cell culture supernatants. The system was developed on the basis of universal primers of Enterovirus and specific primers of vaccinal strain Sabin 1. The achieved results proved that the non-existence of methods of RNA extraction and purification prior to the reaction does not affect the susceptibility and specificity of the system, in the rapid detection of Enterovirus genomes and identification of vaccinal strains of poliovirus.