RESUMEN
To implement effective lumpy skin disease (LSD) control measures, such as timely vaccination, particularly in calves and serological monitoring, it is necessary to evaluate immune response after vaccination, both in adult cattle and in their calves. The aim of this study was to evaluate passive immunity transfer and duration of maternal antibodies against lumpy skin disease virus (LSDV) in calves born to vaccinated cows by two different serological methods. The longitudinal study was carried out on two farms in Serbia where no cases were reported during LSD outbreak in 2016. Fifteen cows on each farm were vaccinated and revaccinated with attenuated vaccine - Neethling strain. A total of 30 cows and 30 calves on both farms were included in the study. Serum samples from cows were collected on calving day and serum samples from their respective calves on days 10, 20, 30, 45, 60, 75, 90, 105 and 120 after birth. Colostrum samples were collected only from 15 cows on one farm. In order to determine the presence of antibodies against LSDV a total of 30 cow sera samples, 15 colostrum samples and 270 calf sera samples were examined by commercial enzyme-linked immunosorbent assay (ELISA) and modified virus neutralization test (VNT). Overall, the performance of both serological tests was very satisfactory. The results of this longitudinal study showed that persistence of passive immunity in calves is less than 4 months, and that most calves are not protected against LSDV at that age. Since the vaccination is the most important control measure against LSDV, the recommended age of six months for vaccination of calves born to vaccinated cows should be reassessed to achieve the most optimal protection against LSD.
Asunto(s)
Anticuerpos Antivirales , Ensayo de Inmunoadsorción Enzimática , Inmunidad Materno-Adquirida , Dermatosis Nodular Contagiosa , Virus de la Dermatosis Nodular Contagiosa , Animales , Bovinos , Dermatosis Nodular Contagiosa/prevención & control , Dermatosis Nodular Contagiosa/inmunología , Dermatosis Nodular Contagiosa/virología , Virus de la Dermatosis Nodular Contagiosa/inmunología , Estudios Longitudinales , Inmunidad Materno-Adquirida/inmunología , Femenino , Anticuerpos Antivirales/sangre , Ensayo de Inmunoadsorción Enzimática/veterinaria , Vacunas Virales/inmunología , Serbia , Pruebas de Neutralización/veterinaria , Vacunación/veterinariaRESUMEN
A single dose protocol of FSH for superstimulation in cattle may improve compliance and superovulatory response. A single subcutaneous (sc) administration of pFSH was efficacious, but response depended on body condition and injection site; the adipose tissue pad behind the shoulder was most efficacious. Inconsistent results in Holsteins were partially overcome by sc administration of 75% of the total pFSH dose behind the shoulder on the first day followed by 25% 48 h later. An alternative would be to combine FSH with polymers that cause it to be released slowly over several days. Hyaluronan is found normally in most animal tissues and is nonreactive when administered parentally. A single intramuscular (im) administration of pFSH in a 2.0% hyaluronan induced a superovulatory response that did not differ from twice daily im administration over 4 d. However, 2.0% hyaluronan was viscous and difficult to mix with FSH. Although solutions of 1.0 and 0.5% hyaluronan were less viscous, they lacked efficacy as a single im administration. However, superovulatory response was high when either 1.0 or 0.5% hyaluronan was used in a two-dose im protocol; two-thirds on the first day and one-third 48 h later. A single im administration of FSH in 0.5% hyaluronan effectively induced superstimulation for OPU in cattle. Successful superovulation in the cow was associated with circulating FSH levels that were similar to endogenous FSH levels prior to follicular wave emergence; however, levels must be maintained above baseline for at least 72 h, or 36 h for OPU. Circulating FSH levels following a single sc administration of 400 mg NIH-FSH-P1 behind the shoulder in beef cows increased to 1.0 or 1.2 ng/mL at 12 h and were back near baseline in approximately 60 h, while FSH levels following im administration of 200 mg NIH-FSH-P1 in 0.5% hyaluronan into Holstein donors reached 1.5 ng/mL at 12 h and returned to baseline in approximately 36 h.
Asunto(s)
Bovinos , Hormona Folículo Estimulante/administración & dosificación , Inducción de la Ovulación/veterinaria , Superovulación , Animales , Femenino , Hormona Folículo Estimulante/sangre , Ácido Hialurónico/administración & dosificación , Inyecciones Subcutáneas , Recuperación del Oocito/métodos , Recuperación del Oocito/veterinaria , Inducción de la Ovulación/métodosRESUMEN
The aim of this study was to determine the level of heavy metals and polycyclic aromatic hydrocarbons (PAHs) in 57 samples of canned tuna, 25 samples of canned sardines, and 16 samples of canned smoked sprouts, collected from supermarkets in Serbia. Heavy metal levels (lead, cadmium, mercury, arsenic, iron, copper, and zinc) were determined using an inductive coupled plasma mass spectrometry method. Sixteen analysed PAHs were determined using a gas chromatography-mass spectrometry method. The trace elements in canned fish were in the range (mg kg-1) of 0.01-0.28 for lead, 0.003-0.08 for cadmium, 0.007-0.64 for mercury, 0.17-3.73 for arsenic, 3.88-52.2 for iron, 0.2-5.95 for copper, and 1.35-44.5 for zinc. Total PAH levels reached 17.67 µg kg-1 in canned tuna and 15.12 µg kg-1 in sardines, whereas in smoked sprouts it amounted to 57.19 µg kg-1. Some of the samples contained cadmium, arsenic, copper, and benzo(a)pyrene above the limits as set by the European and Serbian legislation.
Asunto(s)
Productos Pesqueros/análisis , Contaminación de Alimentos , Conservación de Alimentos , Metales Pesados/química , Hidrocarburos Policíclicos Aromáticos/química , Contaminantes Ambientales/química , Humanos , SerbiaRESUMEN
Three experiments were designed to evaluate the superovulatory response of beef cows following two intramuscular (IM) administrations 48 h apart of Folltropin-V diluted in reduced concentrations of hyaluronan (Split-single IM administrations; Experiment 1-300 mg Folltropin-V on the first day and 100 mg 48 h later; Experiment 2-200 mg Folltropin-V on the first day and 100 mg 48 h later). In Experiments 1 and 2, superovulatory response and ova embryo/embryo production did not differ between donors receiving twice daily IM of Folltropin-V over 4 days and those given a Split-single IM administration of Folltropin-V diluted in 10 mg/mL hyaluronan solution. Experiment 3 compared Split-single IM administration of Folltropin-V diluted in two hyaluronan concentrations (5 or 10 mg/mL) with Folltropin-V diluted in saline and administered twice-daily over 4 days. Beef cows (17 Angus and 12 Simmental) were randomly assigned to one of three treatment groups to be superstimulated three times in a cross-over design, so that all cows received all treatments. A total dose of 300 mg Folltropin-V was divided into twice-daily IM over 4 days, or in two IM treatment 48 h apart (200 mg on first day and 100 mg 48 h later) in the hyaluronan groups. Mean (± SEM) numbers of transferable embryos did not differ among treatment groups (Control: 4.0 ± 0.8; 10 mg/mL hylauronan: 5.0 ± 0.9; 5 mg/mL hyaluronan: 6.1 ± 1.3). We concluded that the Split-single IM administration of Folltropin-V diluted in either concentration of hyaluronan resulted in a comparable superovulatory response to the traditional twice-daily protocol.
Asunto(s)
Hormona Folículo Estimulante/farmacología , Inducción de la Ovulación/veterinaria , Ovulación/efectos de los fármacos , Superovulación/efectos de los fármacos , Animales , Bovinos , Femenino , Hormona Folículo Estimulante/administración & dosificación , Inyecciones Intramusculares/veterinaria , Folículo Ovárico/diagnóstico por imagen , Folículo Ovárico/efectos de los fármacos , Inducción de la Ovulación/métodos , UltrasonografíaRESUMEN
The need to inject FSH twice daily for superstimulation of ovarian follicular development in cattle necessitates frequent attention by farm-personnel and increases the possibility of failures due to mishandling and errors in administration of treatments. A series of three experiments were designed to evaluate the feasibility of superstimulation in beef cattle with a single intramuscular (IM) injection of Folltropin-V diluted in a hyaluronan-based slow-release formulation (SRF). In Experiment 1, cows were assigned to one of three treatment groups to compare two methods of injection as compared to the twice daily IM injection protocol. Superovulatory response of cows (n=6) treated with twice daily IM injections over 4 days (Control) was greater than of cows treated with a single subcutaneous (SC) injection in SRF (n=6), while superovulatory response of cows treated with a single IM injection in SRF (n=6) was intermediate. Experiment 2 was designed to compare two concentrations of SRF (20mg/mL hyaluronan, 100% compared to 10mg/mL hyaluronan, 50%) in a single IM injection protocol. The mean number of corpora lutea (CL) were not significantly different (P≥0.05), but the numbers of total ova/embryos (P<0.05), fertilized ova (P<0.01) and transferable embryos (P<0.001) were greater in cows treated with FSH in 100% SRF (n=20) than cows treated with FSH in 50% SRF (n=20). Experiment 3 was designed to compare superovulatory response in Red Angus donor cows treated with a single IM injection of Folltropin-V diluted in 100% solution of SRF with those treated with the traditional twice-daily IM injection protocol over 4 days. Mean (±SEM) numbers of CL (13.7±1.2 compared to 13.8±1.2), total ova/embryos (12.3±1.5 compared to 13.7±2.1), fertilized ova (7.2±1.1 compared to 8.4±1.4) and transferable embryos (4.9±0.8 compared to 6.4±1.3) were not significantly different between Control (n=29) and Single injection (n=29) groups, respectively. In summary, superstimulation of beef donor cows with a single IM injection of Folltropin-V diluted in 100% solution of SRF resulted in a comparable superovulatory response to the traditional twice-daily IM administration of Folltropin-V diluted in saline over 4 days.
Asunto(s)
Bovinos/fisiología , Hormona Folículo Estimulante/administración & dosificación , Folículo Ovárico/fisiología , Superovulación/fisiología , Animales , Cuerpo Lúteo/diagnóstico por imagen , Cuerpo Lúteo/fisiología , Femenino , Fertilización In Vitro/veterinaria , Inyecciones Intramusculares/veterinaria , Distribución Aleatoria , UltrasonografíaRESUMEN
Escherichia coli O157:H7 remains a threat to humans via cattle-derived fecal contamination of food and water. Preharvest intervention strategies represent a means of reducing the pathogen burden before harvest. In this study, the efficacy of a commercially produced type III secreted protein (TTSP) vaccine was evaluated with the use of a commingled experimental calf infection model (30 placebo-treated animals and 30 vaccinates). The calves were vaccinated on days 0, 21, and 42 and infected with 10(9) colony-forming units (CFU) of E. coli O157 by oral-gastric intubation on day 56. Fecal shedding was monitored daily for 14 d. Serologic assessment revealed a robust immune response to vaccination; the serum titers of antibodies against EspA, Tir, and total TTSPs were significantly higher in the vaccinates than in the placebo-treated animals on days 21, 42, 56, and 70. Significantly less (P = 0.011) of the challenge organism was shed by the vaccinates than by the placebo-treated animals on days 3 to 10. Peak shedding occurred in both groups on days 3 to 6; during this period the vaccinates showed a mean log reduction of 1.4 (P = 0.002) and a mitigated fraction of 51%. The number of animals shedding was significantly lower among the vaccinates compared with the placebo group on days 3 to 6 (P ≤ 0.05), with a mean prevented fraction of 21%. No differences in the duration of shedding were observed. Owing to the low challenge shedding in both groups on days 11 to 14 (mean CFU/g < 10; median = 0), no significant differences were observed. These data indicate that TTSP vaccination had protective effects through significant reductions in the number of animals shedding and the number of challenge organisms shed per animal and provides evidence that TTSP vaccination is an effective preharvest intervention strategy against E. coli O157.
Asunto(s)
Derrame de Bacterias , Enfermedades de los Bovinos/inmunología , Infecciones por Escherichia coli/veterinaria , Escherichia coli O157/fisiología , Proteínas de Escherichia coli/inmunología , Vacunas contra Escherichia coli/inmunología , Vacunación/veterinaria , Animales , Anticuerpos Antibacterianos/sangre , Bovinos , Enfermedades de los Bovinos/sangre , Infecciones por Escherichia coli/inmunología , Infecciones por Escherichia coli/prevención & control , Escherichia coli O157/inmunología , Escherichia coli O157/aislamiento & purificación , Heces/microbiología , Vacunación/métodosRESUMEN
Escherichia coli O157:H7 is an important zoonotic pathogen, causing hemolytic uremic syndrome (HUS). The colonization of cattle and human hosts is mediated through the action of effectors secreted via a type III secretion system (T3SS). The structural genes for the T3SS and many of the secreted effectors are located on a pathogenicity island called the locus of enterocyte effacement (LEE). We cloned and expressed the genes coding for 66 effectors and purified each to measure the cross-reactivity of type III secreted proteins from Shiga toxin-producing Escherichia coli (STEC) serotypes. These included 37 LEE-encoded proteins and 29 non-LEE effectors. The serological response against each protein was measured by Western blot analysis and enzyme-linked immunosorbent assay (ELISA) using sera from rabbits immunized with type III secreted proteins (T3SPs) from four STEC serotypes, experimentally infected cattle, and human sera from six HUS patients. Twenty proteins were recognized by at least one of the STEC T3SP-vaccinated rabbits by Western blotting. Several structural proteins (EspA, EspB, and EspD) and a number of effectors (Tir, NleA, and TccP) were recognized by O26-, O103-, O111-, and O157-specific sera. Sera from experimentally infected cattle and HUS patients were tested using an ELISA against each of the proteins. Tir, EspB, EspD, EspA, and NleA were recognized by the majority of the samples tested. A number of other proteins also were recognized by individual serum samples. Overall, proteins such as Tir, EspB, EspD, NleA, and EspA were highly immunogenic in vaccinated and naturally infected subjects and could be candidates for a cross-protective STEC vaccine.
Asunto(s)
Sistemas de Secreción Bacterianos/inmunología , Proteínas de Escherichia coli/inmunología , Serotipificación , Escherichia coli Shiga-Toxigénica/inmunología , Animales , Proteínas Bacterianas/análisis , Proteínas Bacterianas/inmunología , Bovinos , Reacciones Cruzadas/inmunología , Ensayo de Inmunoadsorción Enzimática , Síndrome Hemolítico-Urémico/inmunología , Síndrome Hemolítico-Urémico/microbiología , Síndrome Hemolítico-Urémico/terapia , Humanos , Inmunoensayo , Conejos , VacunaciónRESUMEN
Shiga toxin producing Escherichia coli (STEC) O26:H11 is an enteric pathogen capable of causing severe hemorrhagic colitis that can lead to hemolytic uremic syndrome. This organism is able to colonize cattle and human intestinal epithelial cells by secreting effectors via a type III secretion system (T3SS). In this investigation, we examined the role of 2 effectors, Tir and NleB, and the structural translocator component EspA in the adherence of STEC to epithelial cells and in the colonization of cattle. Isogenic deletion mutants were constructed and using microscopy and flow cytometry compared to the wild-type strain in their ability to adhere to HEp-2 cells. A competitive assay was also used to measure the capacity of the mutants to colonize the intestinal tract of cattle, where both the mutant and the parental strains were introduced orally at the same time. Genomic DNA was extracted from enriched fecal samples collected at various time points, and quantitative real-time PCR was used to quantify bacteria. A significant reduction in fecal shedding was observed, and adherence to HEp-2 cells was decreased for the tir and espA mutants. Deletion of the nleB gene did not have a significant effect on the adherence of HEp-2 cells; however, in an in vivo model, it strongly reduced the ability of STEC O26:H11 to colonize the bovine intestinal tract.
Asunto(s)
Adhesión Bacteriana , Células Epiteliales/microbiología , Proteínas de Escherichia coli/fisiología , Intestinos/microbiología , Receptores de Superficie Celular/fisiología , Escherichia coli Shiga-Toxigénica/patogenicidad , Factores de Virulencia/fisiología , Animales , Derrame de Bacterias , Bovinos , Línea Celular Tumoral , ADN Bacteriano/genética , Proteínas de Escherichia coli/genética , Heces/microbiología , Citometría de Flujo , Genes Bacterianos , Humanos , Reacción en Cadena de la Polimerasa , Receptores de Superficie Celular/genética , Eliminación de Secuencia , Toxinas Shiga/biosíntesis , Escherichia coli Shiga-Toxigénica/genética , Escherichia coli Shiga-Toxigénica/fisiología , Virulencia/genética , Factores de Virulencia/genéticaRESUMEN
There is continuing need to simplify bovine superovulation protocols without compromising embryo production. The control of follicular wave emergence and ovulation has facilitated donor management, but the most commonly used treatment, oestradiol, cannot be used in many parts of the world and mechanical removal of the dominant follicle is difficult to apply in the field. Other alternatives include gonadotrophin-releasing hormone (GnRH) or LH, but efficacy in groups of randomly cycling animals is variable. Another alternative is to increase the response to GnRH by inducing a persistent follicle and initiating FSH treatments following GnRH-induced ovulation. The number of transferable embryos following superovulation during the first follicular wave did not differ from that achieved 4 days after oestradiol benzoate and progesterone. To further simplify superovulation, FSH has been administered as a single intramuscular injection. Superovulation of beef donors with a single intramuscular injection of Folltropin-V (Bioniche Animal Health, Belleville, ON, Canada) diluted in a slow-release formulation resulted in embryo production comparable to that obtained using the traditional twice-daily protocol. The single intramuscular injection has the potential to reduce labour and handling and may be useful when handling stress is an impediment to success. These alternatives provide ways of facilitating widespread application of embryo transfer technologies.
Asunto(s)
Cruzamiento/métodos , Bovinos/fisiología , Sincronización del Estro/métodos , Superovulación/fisiología , Animales , Transferencia de Embrión/métodos , Transferencia de Embrión/veterinaria , Estradiol/farmacología , Femenino , Hormona Folículo Estimulante/farmacología , Hormona Liberadora de Gonadotropina/farmacologíaRESUMEN
A clinical trial conducted during the summer of 2006 evaluated effects of two- and three-dose regimens of an Escherichia coli O157 vaccine product on the probability of detecting E. coli O157:H7 in feces and colonization of the terminal rectum. The three-dose regimen significantly reduced the probability for cattle to shed E. coli O157:H7 in feces 65% compared to placebo-treated cattle. A dose-effect was demonstrated because a two-dose regimen of the vaccine product was intermediate in effect. These results are consistent with previous estimates of vaccine efficacy against fecal shedding, and agree with our previous finding that efficacy of two or three doses of this vaccine product exhibit a dose-response.
Asunto(s)
Enfermedades de los Bovinos/prevención & control , Infecciones por Escherichia coli/veterinaria , Escherichia coli O157/inmunología , Escherichia coli O157/aislamiento & purificación , Vacunas contra Escherichia coli/administración & dosificación , Animales , Derrame de Bacterias , Distribución Binomial , Bovinos , Enfermedades de los Bovinos/epidemiología , Enfermedades de los Bovinos/microbiología , Células Cultivadas , Recuento de Colonia Microbiana , Dieta , Relación Dosis-Respuesta Inmunológica , Células Epiteliales/microbiología , Infecciones por Escherichia coli/epidemiología , Infecciones por Escherichia coli/microbiología , Infecciones por Escherichia coli/prevención & control , Escherichia coli O157/crecimiento & desarrollo , Heces/microbiología , Mucosa Intestinal/microbiología , Masculino , Vacunación Masiva/veterinaria , Probabilidad , Recto/microbiología , Riesgo , Factores de Tiempo , Resultado del TratamientoRESUMEN
Type III secreted proteins from Escherichia coli O157:H7 are involved in the attachment of the organism to mammalian cells and have been shown to be effective vaccine components capable of reducing colonization of cattle by the organism. In the current study, we used a streptomycin-treated mouse model to evaluate the efficacy of subcutaneous vs intranasal administration of the vaccine. Following immunization, mice were infected with E. coli O157:H7 and feces were monitored for shedding. Immune responses against EspA and Tir were also monitored. Subcutaneous immunization of mice with type III secreted proteins induced significant EspA- and Tir-specific serum IgG antibodies but did not significantly induce any antigen-specific IgA in feces, whereas intranasal immunization elicited significant EspA- and Tir-specific serum IgG antibodies with some animals developing antigen-specific IgA in feces. Only mice that were immunized intranasally with formulations containing mucosal adjuvants, either cholera toxin or CpG-containing oligonucleotides, showed decreased E. coli O157:H7 shedding following experimental infection. Mice immunized subcutaneously with type III secreted proteins did not shed E. coli in feces. These results demonstrate the potential for the use of type III secreted proteins in mucosal vaccine formulations to prevent colonization and shedding of E. coli O157:H7.
Asunto(s)
Infecciones por Escherichia coli/prevención & control , Escherichia coli O157/fisiología , Proteínas de Escherichia coli/administración & dosificación , Vacunas contra Escherichia coli/administración & dosificación , Heces/microbiología , Vacunación , Administración Intranasal , Animales , Anticuerpos Antibacterianos/sangre , Infecciones por Escherichia coli/inmunología , Infecciones por Escherichia coli/microbiología , Escherichia coli O157/inmunología , Proteínas de Escherichia coli/inmunología , Vacunas contra Escherichia coli/inmunología , Femenino , Humanos , Inmunoglobulina A/sangre , Inmunoglobulina G/sangre , Inyecciones Subcutáneas , Ratones , Ratones Endogámicos BALB CRESUMEN
Escherichia coli O157:H7 is an important food- and water-borne pathogen of humans, causing Hemorrhagic Colitis and Haemolytic Uremic Syndrome. Colonization of both cattle and human hosts is mediated through the action of effector molecules secreted via a Type III secretion system, a mechanism shared by other enterohemorrhagic E. coli (EHEC). We recently reported that vaccination of cattle with Type III-secreted proteins (TTSPs) resulted in decreased shedding of the organism following both experimental infection as well as under conditions of natural exposure. In order to extend this to non-O157 EHEC serotypes, we examined the serological cross reactivity of TTSPs of serotypes O26:H11, O103:H2, O111:NM and O157:H7. Western blotting experiments with polyclonal antisera directed against serotype O157:H7 TTSPs suggested that there was significant cross reactivity, although there was limited cross reactivity when two Tir- and EspA-specific monoclonal antibodies were used. Groups of cattle were then vaccinated with TTSPs produced from each of the above serotypes and the magnitude and specificity of the responses were measured. All animals responded well with antibodies to TTSPs of the homologous serotype. However, limited cross reactivity was observed against the others. No cross reactivity was observed against Tir and EspA of serotype O157:H7. These results suggest that vaccination of cattle with TTSPs as a means of reducing the risk of EHEC transmission to humans will induce protection that is serotype specific.
Asunto(s)
Anticuerpos Antibacterianos/inmunología , Escherichia coli Enterohemorrágica/inmunología , Escherichia coli O157/inmunología , Proteínas de Escherichia coli/inmunología , Receptores de Superficie Celular/inmunología , Animales , Anticuerpos Antibacterianos/sangre , Especificidad de Anticuerpos , Adhesión Bacteriana , Bovinos , Enfermedades de los Bovinos/inmunología , Enfermedades de los Bovinos/prevención & control , Línea Celular , Reacciones Cruzadas , Escherichia coli Enterohemorrágica/clasificación , Infecciones por Escherichia coli/inmunología , Infecciones por Escherichia coli/prevención & control , Vacunas contra Escherichia coli/administración & dosificación , Vacunas contra Escherichia coli/inmunología , Humanos , Inmunización , SerotipificaciónRESUMEN
Our objective was to characterize immune parameters in susceptible (SM) and resistant (RM) mares, with and without artificial insemination (AI) and immunomodulation. Eight RM and eight SM were selected based on their reproductive history and functional tests. Both groups of mares were evaluated during three consecutive cycles: Cycle 1, untreated cycle (control); Cycle 2, AI with dead semen; Cycle 3, AI with dead semen and immunomodulation. Endometrial biopsies were taken during the three cycles as follows: Cycle 1--at estrus, when follicles > or =35mm and at diestrus (7+/-1 days after ovulation); Cycle 2--at estrus 24h post-AI, and at diestrus; Cycle 3--at estrus 24h after treatment with a Mycobacterium phlei cell-wall extract (MCWE) and AI, and at diestrus. The mRNA transcription (mRNAT) of IL-8 and IL-10 were determined by real-time PCR. Image analysis of immunohistochemistry slides was performed using digital software (Image-Pro Plus v 5.0; Media Cybernetics); the percentage of stained area was determined for Major Histocompatibility Complex II (MHC-II), polymorphonuclear leukocytes (PMN) and T lymphocytes (TL) on each tissue section. In Cycle 1, SM had significantly higher MHC-II, TL, PMN and IL-8 than RM during estrus (P<0.006, P<0.0005, P<0.05, respectively), while transcription of IL-10 was significantly lower than in RM (P<0.0001). During diestrus, SM had higher levels of TL, PMN and IL-8 than RM (P<0.0001). After AI (Cycle 2), SM had higher levels of IL-8 and lower levels of IL-10 than RM at estrus and no differences were detected for MHC-II, TL and PMN positive cells. During diestrus in the same cycle, all the immune parameters were higher in SM mares (P<0.005, P<0.0004, P<0.0001, P<0.02, respectively). When MCWE was applied at the time of AI (Cycle 3), SM expressed significant higher levels of IL-10 24h after treatment (P<0.005), which were also higher than in the control Cycle 2 or after AI (Cycle 2). However, no significant differences were detected for MHC-II, lymphocytes-PMN or IL-8 between SM and RM during diestrus in Cycle 3. This study showed that SM had higher levels of all immune parameters except IL-10 than RM during Cycle 1. After AI (Cycle 2), the inflammatory condition persisted in SM but not RM mares until day 7 post-ovulation. Following treatment with MCWE at the time of AI (Cycle 3) uterine immunological changes in SM resulted in an endometrial immune environment similar to that found in normal RM.
Asunto(s)
Endometritis/veterinaria , Enfermedades de los Caballos/inmunología , Factores Inmunológicos/farmacología , Inseminación Artificial/efectos adversos , Animales , Pared Celular , Susceptibilidad a Enfermedades , Endometritis/inmunología , Endometritis/prevención & control , Ciclo Estral , Femenino , Genes MHC Clase II , Enfermedades de los Caballos/prevención & control , Caballos , Interleucina-10/genética , Interleucina-10/metabolismo , Interleucina-8/genética , Interleucina-8/metabolismo , Linfocitos/metabolismo , Mycobacterium phlei , Útero/citologíaRESUMEN
A feedlot trial was conducted to assess the efficacy of an Escherichia coli O157:H7 vaccine in reducing fecal shedding of E. coli O157:H7 in 218 pens of feedlot cattle in 9 feedlots in Alberta and Saskatchewan. Pens of cattle were vaccinated once at arrival processing and again at reimplanting with either the E. coli O157:H7 vaccine or a placebo. The E. coli O157:H7 vaccine included 50 microg of type III secreted proteins. Fecal samples were collected from 30 fresh manure patties within each feedlot pen at arrival processing, revaccination at reimplanting, and within 2 wk of slaughter. The mean pen prevalence of E. coli O157:H7 in feces was 5.0%; ranging in pens from 0% to 90%, and varying significantly (P < 0.001) among feedlots. There was no significant association (P > 0.20) between vaccination and pen prevalence of fecal E. coli O157:H7 following initial vaccination, at reimplanting, or prior to slaughter.
Asunto(s)
Vacunas Bacterianas/normas , Enfermedades de los Bovinos/prevención & control , Infecciones por Escherichia coli/veterinaria , Escherichia coli O157/inmunología , Heces/microbiología , Alberta , Animales , Bovinos , Recuento de Colonia Microbiana/veterinaria , Infecciones por Escherichia coli/prevención & control , Escherichia coli O157/aislamiento & purificación , Distribución Aleatoria , SaskatchewanRESUMEN
Porcine reproductive and respiratory syndrome virus (PRRSV) induces apoptosis in cultured cells and in infected tissues of pig, and the GP5 protein was reported to be responsible. To study the role of apoptosis during PRRSV infection, we established a subline of HeLa cells stably expressing the PRRSV GP5 protein under an inducible promoter to prevent cell death that may result from GP5 expression. The expression of GP5 was confirmed upon induction by immunofluorescence, Western blot, and immunoprecipitation. No reduction in cell numbers was observed in the GP5-expressing cells, and neither DNA fragmentation nor strand-breaks were detected in these cells. To examine if bystander cells underwent apoptosis, Marc-145 or HeLa cells were co-cultivated with GP5-expressing cells, and the apoptotic characteristics were examined in the bystanders. Neither DNA laddering, strand-breaks, nor reduction in cell numbers was observed in the co-cultivated cells. Gene expression profiles were examined for both GP5-expressing cells and bystanders by apoptosis-specific macroarrays and gene chip-based microarrays, but no genes related to apoptosis were specifically regulated. Our data suggest that the previously reported GP5-induced apoptosis may be an atypical observation. The GP5-expressing cells were immunoreactive with sera from pigs infected with the North American and European types of PRRSV, which implicates a potential usefulness of these cells as a diagnostic reagent for PRRSV.
Asunto(s)
Virus del Síndrome Respiratorio y Reproductivo Porcino/genética , Proteínas del Envoltorio Viral/metabolismo , Animales , Apoptosis , Expresión Génica , Células HeLa , Humanos , Virus del Síndrome Respiratorio y Reproductivo Porcino/química , Virus del Síndrome Respiratorio y Reproductivo Porcino/inmunología , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/metabolismo , Porcinos , Proteínas del Envoltorio Viral/genéticaRESUMEN
Cattle are an important reservoir of Escherichia coli O157:H7 leading to contamination of food and water, and subsequent human disease. This pathogen colonizes its hosts by producing several proteins such as Tir and EspA that are secreted by a type III secretion system. These proteins play a role in colonization of the intestine, suggesting that they might be useful targets for the development of a vaccine to reduce levels of this organism in cattle. Vaccination of cattle with proteins secreted by E. coli O157:H7 significantly reduced the numbers of bacteria shed in feces, the numbers of animals that shed, and the duration of shedding in an experimental challenge model. Vaccination of cattle also significantly (P=0.04) reduced the prevalence of E. coli O157:H7 in a clinical trial conducted in a typical feedlot setting. This strategy suggests it is possible to vaccinate cattle to decrease the level of E. coli O157:H7 shedding for the purpose of reducing the risk of human disease.
Asunto(s)
Proteínas Bacterianas/inmunología , Vacunas Bacterianas/inmunología , Enfermedades de los Bovinos/prevención & control , Infecciones por Escherichia coli/prevención & control , Infecciones por Escherichia coli/veterinaria , Escherichia coli O157/inmunología , Animales , Bovinos , Enfermedades de los Bovinos/inmunología , Química Farmacéutica , Reservorios de Enfermedades , Infecciones por Escherichia coli/microbiología , Escherichia coli O157/genética , Heces/microbiología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , VacunaciónRESUMEN
Endometrial mRNA expression of the pro-inflammatory cytokines interleukin-1beta (IL-1beta), interleukin-6 (IL-6), and tumor necrosis factor alpha (TNF-alpha) was assessed in mares resistant (RM) or susceptible (SM) to persistent post-breeding endometritis (PPBE). Eight RM and eight SM, were selected based on reproductive records and functional tests out of a herd of 2,000 light cross-type mares. Three experiments were done to study transcription patterns in (i) basal conditions; (ii) after artificial insemination (AI); and (iii) after administration of an immunomodulator at time of artificial insemination. Endometrial biopsies were taken during consecutive cycles: (i) at estrus, when follicles reached 35 mm and at diestrus (7 +/- 1 days after ovulation); (ii) at 24 h post-AI, with dead semen (estrus) and in diestrus; (iii) at 24 h after treatment with a Mycobacterium phlei cell-wall extract (MCWE) preparation and AI (with dead semen), and at diestrus. mRNA expression was quantitated by real time PCR. Under basal conditions, SM had significantly higher mRNA expression of all cytokines in estrus and of IL-1beta and TNF-alpha in diestrus, compared to RM. After AI, there were no differences between RM and SM in estrus; however, mRNA expression for all three pro-inflammatory cytokines was higher than under basal conditions. In diestrus, RM showed significantly lower IL-1beta and TNF-alpha mRNA expression than SM. When MCWE was administered at time of AI, no differences between cytokine induction from RM and SM were found. Globally, mRNA expression for all three cytokines correlated well among themselves when expression was high. The present study showed that (i) in basal conditions RM had lower mRNA expression of pro-inflammatory cytokines than SM with no effect of estrous cycle; (ii) AI upregulated mRNA expression for all three cytokines in both RM and SM, with persistance in diestrus in the latter; (iii) treatment with MCWE at time of AI down-regulated mRNA expression of IL-1 with significant effects in SM which behaved like RM. Immunomodulation with MCWE could be of help in restoring homeostatic local inflammatory mechanisms, thus assisting in the prophylaxis of post-breeding endometritis in mares.