Delayed-type hypersensitivity skin test against Neospora caninum in heifers with undetectable specific antibodies.

Delayed-type hypersensitivity (DTH) has been used in human and veterinary medicine as a skin testing for evaluating in vivo cell-mediated immune responses (CMIR). Whereas CMIR is a key process to control intracellular pathogens, its value at identifying cattle exposed to the abortigenic intracellular coccidian parasite Neospora caninum is unknown. In this work, we have evaluated a DTH skin testing in cattle exposed to N. caninum and still seronegative. Female calves were experimentally sensitized by subcutaneous (SC) inoculation with live tachyzoites of N. caninum (NC-Argentina LP1) in sterile phosphate-buffered saline (PBS) (group A; n: 8) whereas other calveswere mock-sensitized with PBS (group B; n: 6). Two DTH skin tests were performed by intradermal inoculation with a soluble lysate of N. caninum tachyzoites (NC-Argentina LP1) in the neck region at 60d and 960 d after sensitization. Skinfold thickness at the intradermal inoculation site was measured at 0, 24, 48 h post each DTH skin test and skin biopsies taken for microscopic evaluation. Specific N. caninum antibodies kinetics was evaluated all throughthe experiment. We found that whereas N. caninum specific antibodies remained below the ELISA cut-off, a distinctive skinfold thickness increase was detected in sensitized animals (group A) at the DTH skin test site, showing induration, swelling and inflammatory infiltration. Mock sensitized animals (group B) showed no skinfold thickness growth and lacked specific antibody response. Thus, N. caninum DTH skin testing could be a useful diagnostic tool for the detection of CMIR during N. caninum infection in non-humoral responders.

Cathelicidin bovine myeloid antimicrobial peptide (BMAP) 28 is involved in the inflammatory response against alpha-herpesviruses in the bovine nervous system.

The role of the local innate immune response in the neuropathogenesis of bovine herpesvirus (BoHV) type 1 and 5 remains largely unknown. This study determined the gene transcriptional expression of relevant bovine cathelicidins, TNFα and IFNß in the nervous system of experimentally-infected cattle during the different stages of BoHV-1 and BoHV-5 infectious cycle. We studied the modulation of bovine myeloid antimicrobial peptide (BMAP) 27 and 28 by alpha-herpesviruses during acute infection of the central nervous system (CNS). However, BMAP28 was the main cathelicidin modulated. BoHV-5 supressed BMAP28 expression mainly in frontal cortex and cervical medulla whereas BoHV-1 slightly induced the expression of cathelicidins in the olfactory and posterior cortex. The differences in the regulation of the innate response are likely related to distinct replication rates of both alpha-herpesviruses in the CNS. During latency and reactivation, BoHV-1 and -5 decreased BMAP28 and BMAP27 expression, accompanied by high levels of TNFα and IFNß transcripts in the posterior brain region and medulla during BoHV reactivation. In terms of cytokines, a remarkably overexpression of IFNß was induced by BoHV-5 (133.8-fold). In trigeminal ganglion (TG) both alpha-herpesviruses induced cathelidicins gene expression at all stages of the infection cycle, while only acute BoHV-5 infection increased TNFα (129-fold) mRNA levels. This study suggests that the pronounced downregulation of BMAP28 in BoHV-5-acutely-infected CNS is due to a decreased immune stimulation during viral infection, favouring its establishment in the CNS with a low replication rate until latency. Thus, cathelicidins, together with IFNß and TNFα, are differentially regulated by BoHV-5 and BoHV-1 infections and this regulation is dependent on the stage of virus infection in the bovine nervous system.

Modulation of cathelicidins, IFNß and TNFα by bovine alpha-herpesviruses is dependent on the stage of the infectious cycle.

Production of antimicrobial peptides cathelicidins, interferons and cytokines is an important feature in airway epithelial host defense. The innate immune response to alpha-herpesvirus infection at the sites of primary replication has not been fully studied. Thus, the aim of this study was to determine the expression of innate immune components, cathelicidins, IFNß, TNFα and TNF receptors (TNFRI and TNFRII) during acute infection and reactivation of bovine herpesvirus type 1 (BoHV-1) and 5 (BoHV-5) in the respiratory tract and lymphoid tissue of their natural host. We found that BoHV infection modulates mainly the expression of BMAP28, a key cathelicidin in cattle. It was downregulated by both viruses in retropharyngeal lymph nodes of acutely infected-calves, and it was accompanied by a lower expression of IFNß, TNFα and TNFRI. BoHV-5 showed a pronounced role in the downregulation of BMAP28, even in nasal mucosa and lung. However, during reactivation, BoHV-5 upregulated both BMAP28 and IFNß in retropharyngeal lymph nodes. Acute replication induced also TNFα mRNA and protein synthesis, and expression of TNFRI and II was positively regulated during both acute infection and reactivation, particularly in the trachea. Moreover, BMAP27 was detected during BoHV-1 reactivation suggesting a potential role at this stage. Thus, cathelicidins are implicated in alpha-herpesvirus infections of the bovine respiratory system and the response is distinct during BoHV-1 and BoHV-5 acute infection and reactivation. This demonstrates that these viruses modulate differentially the components of innate immune response, possibly influencing their pathogenesis. This study provides an initial pilot analysis of factors that might be implicated in alpha-herpesvirus infection of the bovine respiratory system.

Utilidad clínica de la procalcitonina en gastroenteritis aguda / Use of procalcitonin as diagnostic marker in acute gastroenteritis

Introducción: No hay ningún marcador hematológico que diferencie con seguridad entre gastroenteritis aguda (GEA) bacteriana y no bacteriana. Nuestro objetivo fue evaluar la procalcitonina (PCT) como marcador de GEA de origen bacteriano y analizar su correlación con el ingreso hospitalario. Pacientes y Método: Estudio prospectivo de niños diagnosticados de GEA en el departamento de Urgencias durante un período de 7 meses que requirieron de analítica sanguínea y muestras de heces. Se analizaron variables epidemiológicas, clínicas y analíticas. Se excluyeron pacientes con enfermedad digestiva crónica, diarrea prolongada, inmunodeficiencia o tratamiento antibiótico previo. El estudio fue aprobado por el Comité de Ética y se solicitó consentimiento informado. Resultados: Se analizaron 45 pacientes. Los niños con GEA bacteriana tenían mayor edad (p = 0,027), mayor mediana de PCT y proteína C reactiva (PCR) (p = 0,001). Los valores de PCT y PCR que mejor discriminaron la etiología bacteriana fueron PCT > 0,5 mg/L (sensibilidad: 64,3 por ciento, especificidad: 83,9 por ciento, cociente de probabilidad positivo (CPP): 4) y PCR > 3 mg/dL (sensibilidad: 78,6 por ciento, especificidad: 90,3 por ciento, CPP: 8). No se encontró asociación entre la elevación de dichos marcadores y una mayor probabilidad de hospitalización. Conclusión: La procalcitonina, al igual que la PCR, se eleva en gastroenteritis bacterianas (p = 0,001), no siendo estos marcadores predictores de hospitalización.

Introduction: There is no hematological marker that reliably differentiates between bacterial and nonbacterial acute gastroenteritis (AGE). The objective of this study is to evaluate procalcitonin (PCT) as a marker for bacterial AGE and analyze its relationship with hospital admission. Patients and Method: A prospective study of children diagnosed with AGE was conducted at the emergency room during a period of seven months, which required blood and stool samples. Epidemiological, clinical and analytical variables were analyzed. Patients with chronic digestive disease, prolonged diarrhea, immunodeficiency or prior antibiotic treatment were excluded. The study was approved by the Ethics Committee and an informed consent was requested. Results: 45 patients were analyzed. Children with bacterial GEA were older (p = 0.027) and presented higher median PCT and C-reactive protein concentrations (CRP) (p = 0.001). The PCT and CRP values that best discriminated bacterial infection were PCT > 0.05 mg/L (sensibility 64.3 percent specificity 83.9 percent, positive probability coefficient (PPC): 4), and CRP > 3mg/dL (sensibility 78.6 percent, specificity 90.3 percent, PPC: 8). No association between the elevation of these markers and higher hospitalization probability was found. Conclusion: Procalcitonin, like CRP, is elevated in bacterial gastroenteritis (p = 0.001), but these markers are not a predictor of hospitalization.

[Use of procalcitonin as diagnostic marker in acute gastroenteritis]. / Utilidad clínica de la procalcitonina en gastroenteritis aguda.

INTRODUCTION: There is no hematological marker that reliably differentiates between bacterial and nonbacterial acute gastroenteritis (AGE). The objective of this study is to evaluate procalcitonin (PCT) as a marker for bacterial AGE and analyze its relationship with hospital admission. PATIENTS AND METHOD: A prospective study of children diagnosed with AGE was conducted at the emergency room during a period of seven months, which required blood and stool samples. Epidemiological, clinical and analytical variables were analyzed. Patients with chronic digestive disease, prolonged diarrhea, immunodeficiency or prior antibiotic treatment were excluded. The study was approved by the Ethics Committee and an informed consent was requested. RESULTS: 45 patients were analyzed. Children with bacterial GEA were older (p=0.027) and presented higher median PCT and C-reactive protein concentrations (CRP) (p=0.001). The PCT and CRP values that best discriminated bacterial infection were PCT≥0.05 mg/L (sensibility 64.3%, specificity 83.9%, positive probability coefficient (PPC): 4), and CRP≥3 mg/dL (sensibility 78.6%, specificity 90.3%, PPC: 8). No association between the elevation of these markers and higher hospitalization probability was found. CONCLUSION: Procalcitonin, like CRP, is elevated in bacterial gastroenteritis (p=0.001), but these markers are not a predictor of hospitalization.

Lectin binding patterns and immunohistochemical antigen detection in the genitalia of Tritrichomonas foetus-infected heifers.

Heifers inoculated intra-vaginally with Tritrichomonas foetus were examined after long-term infection (70 days) and short-term infection (20 days) by lectin-histochemical, immunohistochemical and cultural techniques. The organism was recovered from the genital tract and T. foetus antigens were detected immunohistochemically in the lumina of uterine glands and cytoplasm of vaginal subepithelial macrophages. An increase of galactosylated residues (galactose and N-acetyl galactose), binding to PNA, was observed in the genital epithelium (vagina, uterus and oviduct) from infected animals. In the oviductal epithelium of short- but not long-term infected heifers, mannose (binding to Con A) was detected, suggesting that the persistent presence of T. foetus and its virulence factors or inflammatory processes result in a change in the glycoproteins of the epithelial surface. The findings have implications for the adhesion of T. foetus to cells and for the pathogenesis of bovine trichomonosis.

Effect of two commercial vaccines to Campylobacter fetus subspecies on heifers naturally challenged.

Efficacy of two commercial vaccines containing Campylobacter fetus subspecies on heifers naturally challenged by service with an infected bull was tested. Sixteen heifers were vaccinated parentally two times with 3 weeks as interval, eight with commercial vaccine A and the other eight with commercial vaccine B. Eight other heifers were used as unvaccinated controls. Forty days after the first vaccine dose, the heifers were served by an infected bull during 60 days. Measure of systemic immune response and identification of the microorganism from genital secretions by culture and immunofluorescent antibody test (IFAT) were done. Vaccinated and control heifers had a poor reproductive performance (pregnancy rates were 2/8, 3/8 and 0/8 in groups A, B and C, respectively) and were infected by both methods during breeding time and after it. Moreover, one heifer in the groups B and C remained infected until 300 days post-breeding time. Neither vaccinated nor control heifers had an important increment of systemic antibody level. Only, they had a slight increment of antibody level after the breeding period and it may be because of natural stimulus by the infected bull during the copula. Culture and IFAT yielded high correlation on identification of C. fetus subspecies.

Two-step (culture and PCR) diagnostic approach for differentiation of non-T. foetus trichomonads from genitalia of virgin beef bulls in Argentina.

Preputial fluids from 567 virgin Angus and Hereford bulls, 1-2 years old, were inoculated into Sutherland medium, and approximately 8.4% produced cultures with a protozoan suggestive of Tritrichomonas foetus. Under brightfield microscopy, large numbers of single-celled motile organisms with multiple anterior flagellae, a posterior flagellum, axostyle, and a visible undulating membrane were detectable. Motility was jerky and rolling, as described for T. foetus. Air-dried smears of cultures stained with Giemsa or Diff-Quick + iodine revealed an organism similar to T. foetus, although somewhat more rounded. Several organisms appeared to have four anterior flagellae. Scanning electron microscopy (5000x) of representative samples revealed four anterior flagellae on most organisms, and an axostyle that was consistently longer than that seen in T. foetus. Using pan-trichomonal primers and T. foetus-specific primers in a polymerase chain reaction (PCR) assay, amplification products of 372bp were detected in all virgin bull isolates, but only with the pan-trichomonal primers. Positive control isolates of T. foetus yielded amplification products of the expected size (372 and 347bp) with the two sets of primers, respectively. We conclude that these protozoa are not T. foetus, and note the similarity of these findings with those reported earlier in North American beef cattle. Because in several countries there is no legal treatment for bovine trichomonosis, veterinarians recommend slaughter of bulls with positive preputial cultures. The existence of easily mis-identified non-T. foetus trichomonads in the bovine prepuce suggests that the current "gold standard" diagnostic test (culture of preputial scrapings or washings) should be augmented with a more specific confirming test, such as the PCR employed in this study.

Experimental infection with Tritrichomonas suis in heifers.

Nine heifers were intravaginally challenged with 9.3x10(6) Tritrichomonas suis reference strains. Vaginal mucus and serum samples were collected weekly 4 weeks post-inoculation. Vaginal mucus was cultured for T. suis and sera was tested by ELISA against whole cell antigens for T. suis and Tritrichomonas foetus. All vaginal mucus cultures were T. suis-negative during the experiment. ELISA values for both antigens were similar and differences were not significant (P>0.05). Positive control serum samples from one heifer vaccinated against T. foetus showed anti-T. suis ELISA values. We concluded that T. suis intravaginal inoculation induced a low level of serum immune response in heifers measured by ELISA and both protozoa probably share a common antigen. However, under the experimental conditions of this trial, colonization of the heifers' genital tract was not possible in any of the nine animals.

Cervical cerclage: an alternative for the management of placenta previa?

OBJECTIVE: Our purpose was to determine whether cervical cerclage reduces the maternal and neonatal morbidity in women with placenta previa. STUDY DESIGN: Thirty-nine pregnant women with an initial diagnosis of placenta previa at 24 to 30 weeks' gestation were randomly assigned to cervical cerclage (n = 19) or conservative management (n = 20). Subjects were followed up until delivery. Primary outcome measure was gestational age at delivery. Secondary outcome measures were prolongation of pregnancy, number of patients bleeding after being randomly assigned, units of blood transfused, birth weight, hospital stay and costs, and admission to neonatal intensive care unit. Statistical significance was calculated by the Student t test, Fisher's exact probability test, and the chi2 with Yates' correction factor. RESULTS: No statistically significant differences were observed between the two groups studied. CONCLUSION: Cervical cerclage does not appear to be an adequate alternative for the management of placenta previa.

Severe preeclampsia and postpartum eclampsia associated with placenta previa and cesarean and hysterectomy: a case report.

Several prior studies have demonstrated that the presence of placenta previa protects against the development of preeclampsia. Also, there are no reported cases of eclampsia following cesarean section and hysterectomy. However, we encountered a patient with total placenta previa and accreta who developed severe antepartum preeclampsia followed by the onset of eclamptic seizures after a cesarean section and a subtotal hysterectomy. This case confirms that severe preeclampsia may occur in patients with placenta previa and demonstrate that severe preeclampsia may occur after placental removal and even after subtotal hysterectomy, which removed most of the trophoblastic tissue.

Characteristics of the spontaneous milk ejecting activity occurring during human lactation.

Continuous recordings of intramammary pressure performed in 158 normal lactating women, and 22 women during spontaneous labor were retrospectively analyzed in order to characterize spontaneous milk ejecting activity (SA). SA appeared as spurts of contraction waves as early as the first post-partum day and throughout the first two months of lactation. The spurts of SA differed from reflex milk ejection evoked by baby's suckling. It showed two different patterns: a) single contraction waves with a rapid rise time, a slow decay time, high amplitude and a short duration; b) multiple contraction waves with a longer rise and decay times, progressively decreasing intensity and a large duration depending on the number of contractions per spurt. The multiple pattern occurred in 34.2% (41/120) of patients at 2.7 +/- 0.1 days of post-partum and increased to 84.3% (59/70) at 34.5 +/- 1.2 days. Accordingly, duration and the area under contraction curves increased throughout lactation. Mammary gland sensitivity to exogenous oxytocin (OT) did not change during the 2 months studies. SA was significantly greater in full nursing mothers than in the partially nursing ones. The existence of a system for milk-ejection different from baby's suckling was further described. A neuroendocrine mechanism and the release of increasing amounts of OT are suggested as important factors in the control of this phenomenon.

Correlacion entre la duracion clinica y la intensidad de las contracciones uterinas en el parto humano. / Correlation between the clinical length of time intensity of the uterine contractions in human labor

La correlacion entre la duracion clinica y la intensidad de las contracciones uterinas, no se ha establecido hasta ahora.En este trabajo se estudia ese parametro y se informan los resultados obtenidos midiendo como duracion clinica, la que se puede cuantificar cuando la contraccion alcanza el valor de 20 mm Hg. La duracion promedio en 55 partos estudiados fue 39.4 +/- 0.87 segundos, valor similar al de la intensidad que fue 39.4 +/- 0.87 mm Hg. El coeficiente de correlacion fue alto (0.81) y la ecuacion de regresion (y = 20 427 + 0.73X) permitio construir una recta donde se observo que los valores normales de intensidad se corresponden con los de duracion clinica. Ademas, como ocurre con la intensidad, la duracion clinica aumenta significativamente al final del primer periodo del parto. Las contracciones inducidas con oxitocina presentan el mismo comportamiento de las espontaneas; en cambio, las contraciones incoordinadas no muestran las relaciones descritas atras

Reactividad del utero a la oxitocina exogena en la hipertension arterial inducida por el embarazo. / Reactivity of uterus to exogenous oxytocin in arterial hypertension induced by pregnancy

La reactividad del utero humano a la oxitocina no se ha estudiado suficientemente en los casos de hipertension inducida por el embarazo. Ademas, los resultados que hasta ahora se conocen son contradictorios. En este articulo se presentan los hallazgos de un estudio controlado que se efectuo en el Hospital Universitario del Valle, los cuales demuestran que no hay diferencias en la capacidad de respuesta del miometrio, al termino del embarazo, entre los controles y las pacientes con pre-eclampsia severa, tanto antes como despues del tratamiento del sindrome con sulfato de magnesio. Se discute la significacion de estos resultados