Experimental infection with Tritrichomonas suis in heifers

Nine heifers were intravaginally challenged with 9.3 × 106 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. © 2001 Elsevier Science B.V.Fil: Cobo, Eduardo Ruben. Instituto Nacional de Tecnología Agropecuaria; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Cano, Dora. Instituto Nacional de Tecnología Agropecuaria; ArgentinaFil: Campero, Carlos Manuel. Instituto Nacional de Tecnología Agropecuaria; Argentin

Nuevos aspectos inmunológicos y vacunales de la tricomoniasis bovina

En la presente revisión se describen aspectos generales de la enfermedad y los últimos avances científicos logrados en el área de la Tricomoniasis bovina. Se consideran aspectos vinculados a la patogenia, abarcando la relación molecular parásito-célula huésped y los mecanismos inmunes humorales y celulares involucrados en dicha enfermedad. Además se menciona el desarrollo de nuevas técnicas diagnósticas y los avances en el desarrollo y evaluación de vacunas contra Tritrichomonas foetus (T. foetus).Fil: Cobo, Eduardo Ruben. Instituto Nacional de Tecnología Agropecuaria; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Campero, Carlos Manuel. Instituto Nacional de Tecnología Agropecuaria; Argentin

Lectin binding pattern 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.Fil: Cobo, Eduardo Ruben. Instituto Nacional de Tecnología Agropecuaria; ArgentinaFil: Campero, Carlos Manuel. Instituto Nacional de Tecnología Agropecuaria; ArgentinaFil: Gimeno, Eduardo Juan. Universidad Nacional de La Plata; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Barbeito, Claudio Gustavo. Universidad Nacional de La Plata; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

Heifers immunized with whole-cell and membrane vaccines against Tritrichomonas foetus and naturally challenged with an infected bull

The performance of a whole-cell vaccine and the other vaccine with cellular membranes of Tritrichomonas foetus applied to heifers naturally challenged by mating with an infected bull was determined. Forty heifers were divided into three groups: a control group (n = 16) without immunizing, another group (n = 12) immunized with whole cells (108/dose) and a third group (n = 12) immunized with cellular membranes (300 μg of membranes/dose protein). The females were subcutaneously vaccinated at 3-week on two occasions and received a third intravaginal booster dose. After 3 weeks of the last vaccinal doses, the heifers were served by a T. foetus infected bull over 90-day period. The mean duration of infection for membrane-vaccinated heifers was 60 days ±25, compared with 63 days ±35.8 of infection for whole-cell-vaccinated heifers and 79 days ±41.3 for control heifers. Calving rates were 6/12 for membrane-vaccinated heifers, 3/12 for whole-cell-vaccinated animals, and 2/16 for control animals. Fetal mortality rates were 3/12 for membrane-vaccinated animals, 4/12 for those vaccinated with whole cells and 10/16 for control animals. These reproductive parameters were significantly different (P < 0.05) between heifers vaccinated with membranes and control heifers. The hemolytic test and enzyme-linked immunoabsorbent assay (ELISA) with T. foetus antigen showed that serum immunoglobulins peaked before and during the breeding period. The heifers vaccinated with membranes developed an important response during the critical period of fetal loss, second and third month of the breeding time, and another month after the same period. The ELISA method was more sensitive and more reliable than the hemolytic test for the evaluation of the systemic immune response in females infected and/or vaccinated with T. foetus.Fil: Cobo, Eduardo Ruben. Instituto Nacional de Tecnología Agropecuaria; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Cano, Dora. Instituto Nacional de Tecnología Agropecuaria; ArgentinaFil: Rossetti, Osvaldo Luis. Instituto Nacional de Tecnología Agropecuaria; ArgentinaFil: Campero, Carlos Manuel. Instituto Nacional de Tecnología Agropecuaria; Argentin

Immunization in heifers with dual vaccines containing Tritrichomonas foetus and Campylobacter fetus antigens using systemic and mucosal routes

Vaccines against both bovine venereal campylobacteriosis and trichomonosis were tested. Heifers were assigned to three groups. Groups 1 (n = 21 heifers) and group 2 (n = 20) received a commercial or experimental vaccine, respectively, containing both Campylobacter fetus and Tritrichomonas foetus antigens. Group 3 (n = 21) received adjuvant alone. Preparations were injected SQ in groups 1 and 3 at days -60 and -30 (day 0 was considered the first day of a 90-day breeding period), and in group 2 SQ at days -30 and +11 and into the vaginal submucosa at day -9. Heifers were exposed to two pathogen-infected bulls for 90 days (from day 0 to day +90); furthermore, half of the heifers in each group were challenged at day +39 by an intravaginal instillation of C. fetus venerealis and T. foetus. Pregnancy diagnosis, vaginal culture, and determination of systemic IgG for both organisms were performed. Compared to controls, vaccinated heifers resisted or quickly cleared both pathogens, had a higher pregnancy rate and a higher systemic immune response during and after the breeding period. Overall, the experimental vaccine was superior to the commercial vaccine (groups 2 and 1, respectively). In conclusion, an experimental vaccine containing both C. fetus and T. foetus antigens, given both SQ and intravaginal immediately before breeding and early in the breeding season, yielded superior protection for heifers exposed to bulls harboring C. fetus and T. foetus.Fil: Cobo, Eduardo Ruben. Instituto Nacional de Tecnología Agropecuaria; ArgentinaFil: Morsella, Claudia. Instituto Nacional de Tecnología Agropecuaria; ArgentinaFil: Cano, Dora. Instituto Nacional de Tecnología Agropecuaria; ArgentinaFil: Cipolla, Ana. Instituto Nacional de Tecnología Agropecuaria; ArgentinaFil: Campero, Carlos Manuel. Instituto Nacional de Tecnología Agropecuaria; Argentin

Failure to establish infection with Tetratrichomonas sp. in the reproductive tracts of heifers and bulls

Experimental infection of the reproductive tracts of heifers and bulls with Tetratrichomonas sp. isolated from preputial smegma of virgin bulls was attempted. Nine heifers and four bulls were challenged by inoculation of 7×106 Tetratrichomonas sp. into the vaginal lumen and preputial cavity, respectively. Vaginal mucus and preputial smegma samples were collected and cultured for Tetratrichomonas sp. Heifers were slaughtered in groups of three at 2, 9 and 21 days after inoculation. Two heifers and two bulls infected with Tritrichomonas foetus and two uninfected heifers were used as controls for the model infection. Tetratrichomonas sp. were only isolated in vaginal mucus of 7/9 inoculated heifers at 6 h post-inoculation, and genital secretions taken at slaughter time from vagina, uterus and oviduct were cultural negative. Bulls challenged with Tetratrichomonas sp. remained cultural negative. Since Tetratrichomonas sp. survived only a few hours in the female genitalia and did not survive in the male genitalia after experimental challenge, Tetratrichomonas sp. did not colonize the genital tract. These were likely trichomonads from the digestive tract. Collection of clean samples without fecal contamination from the reproductive tract is proposed as a measure to avoid Tetratrichomonas sp. transitory genital infection.Fil: Cobo, Eduardo Ruben. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Canton, German. Instituto Nacional de Tecnología Agropecuaria; ArgentinaFil: Morrell, Eleonora Lidia. Instituto Nacional de Tecnología Agropecuaria; ArgentinaFil: Cano, Dora. Instituto Nacional de Tecnología Agropecuaria; ArgentinaFil: Campero, Carlos Manuel. Instituto Nacional de Tecnología Agropecuaria; Argentin

Cathelicidin modulates synthesis of Toll-like Receptors (TLRs) 4 and 9 in colonic epithelium

Cathelicidin are innate antimicrobial peptides with broad immunomodulatory functions; however, their role in regulating intestinal defenses is not well characterized. This study aimed to investigate the role of cathelicidin modulating expression of Toll-like receptors (TLRs) 4 and 9 in colonic epithelium in response to bacterial patterns. We demonstrated herein that intestinal epithelial cells, when primed by bacterial lipopolysaccharide (LPS), responded to cathelicidin by increased transcription and protein synthesis of TLR4. This cathelicidin-induced response required the interaction of LPS-TLR4 and activation of MAPK signalling pathways. However, cathelicidin blocked TLR9 responses induced by TLR9 ligand CpG oligodeoxynucleotide (CpG ODN) in these colonic epithelial cells. Modulations of TLRs triggered by cathelicidin in intestinal epithelium occurred mainly in the apical compartment of intestinal cells. Activation of TLR4 by ligands in combination with cathelicidin promoted CXCL8 chemokine secretion and epithelial antimicrobial defenses against Escherichia coli. We concluded that cathelicidin selectively modulated synthesis of TLR4 and 9 in intestinal epithelium, but only when cells were exposed to virulence factors, mostly from apical surfaces. Enhanced TLR4 expression promoted by cathelicidin in intestinal epithelium may be crucial for controlling enteric infectious diseases.Fil: Marin, Maia Solange. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Holani, Ravi. University of Calgary; CanadáFil: Shah, Chaitanya B.. University of Calgary; CanadáFil: Odeón, Anselmo Carlos. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires Sur. Estación Experimental Agropecuaria Balcarce; ArgentinaFil: Cobo, Eduardo Ruben. University of Calgary; Canadá. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

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 IFNβ 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, BMAP 28 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 IFNβ transcripts in the posterior brain region and medulla during BoHV reactivation. In terms of cytokines, a remarkably overexpression of IFNβ 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 IFNβ 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.Fil: Burucúa, Mercedes María. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires Sur. Estación Experimental Agropecuaria Balcarce; ArgentinaFil: Perez, Sandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; ArgentinaFil: Odeón, Anselmo Carlos. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires Sur. Estación Experimental Agropecuaria Balcarce; ArgentinaFil: Cobo, Eduardo Ruben. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. University of Calgary; CanadáFil: Quintana, Silvina. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Departamento de Biología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Marin, Maia Solange. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires Sur. Estación Experimental Agropecuaria Balcarce; Argentin

Estimación de las pérdidas de la precipitación en una cuenca hidrográfica urbana de la Ciudad de Cuenca

La ciudad de Cuenca en Ecuador, enfrenta un permanente proceso de urbanización de su territorio, provocando considerable alteración en la respuesta hidrológica de la cuenca hidrográfica ante las lluvias. Entre las alteraciones que sufre una cuenca urbanizada se tiene la variación de la pérdida de la lluvia en la cuenca (abstracciones), existiendo diversas metodologías para su estimación. Entre ellas se cuenta con el método de Coeficiente de escorrentía volumétrico Cv. Con el presente trabajo, a través de la metodología señalada se plantea estimar las pérdidas o abstracciones que experimenta una cuenca urbana en la ciudad de Cuenca mediante el análisis de información recopilada entre Marzo y Mayo del 2017. Los resultados que se obtuvieron indican que para precipitaciones con intensidades que varían entre 4.8 y 38.4 mm/h las pérdidas en la cuenca se encuentran entre el 18% y 60%.Cuenc

Characteristic pro-inflammatory cytokines and host defence cathelicidin peptide produced by human monocyte-derived macrophages infected with Neospora caninum

Neospora caninum is a coccidian intracellular protozoan capable of infecting a wide range of mammals, although severe disease is mostly reported in dogs and cattle. Innate defences triggered by monocytes/macrophages are key in the pathogenesis of neosporosis, as these cells are first-line defenders against intracellular infections. The aim of this study was to characterize infection and innate responses in macrophages infected with N. caninum using a well-known cell model to study macrophage functions (human monocyte THP-1 cells). Intracellular invasion of live tachyzoites occurred as fast as 4 h (confirmed with immunofluorescence microscopy using N. caninum-specific antibodies). Macrophages infected by N. caninum had increased expression of pro-inflammatory cytokines (TNFα, IL-1β, IL-8, IFNγ). Interestingly, N. caninum induced expression of host-defence peptides (cathelicidins), a mechanism of defence never reported for N. caninum infection in macrophages. The expression of cytokines and cathelicidins in macrophages invaded by N. caninum was mediated by mitogen-activated protein kinase (MEK 1/2). Secretion of such innate factors from N. caninum-infected macrophages reduced parasite internalization and promoted the secretion of pro-inflammatory cytokines in naïve macrophages. We concluded that rapid invasion of macrophages by N. caninum triggered protective innate defence mechanisms against intracellular pathogens.Fil: Boucher, E.. University of Calgary; CanadáFil: Marin, Maia Solange. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata; ArgentinaFil: Holani, R.. University of Calgary; CanadáFil: Young Speirs, M.. University of Calgary; CanadáFil: Moore, Dadin Prando. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata; ArgentinaFil: Cobo, Eduardo Ruben. University of Calgary; Canad