Genetic parameters for mouth size and their influence on growth traits in pasture-raised Nelore cattle.

Evaluating traits that allow breeders to increase production efficiency in beef production systems is important. The mouth size (MS) score is a trait easily measured and implemented by breeders. Bite size is related to MS in beef cattle and is a determinant of daily feed intake of pasture-raised animals, influencing their growth. The aim of this study was to estimate genetic parameters for MS, weaning weight (WW) and postweaning weight gain (PWG) of Nelore cattle and to evaluate the influence of the interaction between MS and WW on PWG. Phenotypic records of 134,282 Nelore animals born between 1995 and 2019 were used. Variance components were estimated using multitrait animal model with the Bayesian method. The model included the contemporary group as fixed effect, age at measurement of the trait as linear covariate, and direct additive genetic and residual effects as random effects. For WW, random maternal and maternal permanent environmental effects were added to the model. A Bayesian approach was used to analyze the interaction between WW clusters and MS. The heritability estimates were 0.24, 0.15, and 0.23 for MS, WW, and PWG, respectively. The genetic correlation between variables studied ranged from 0.24 to 0.46. The results suggest that animals with a larger mouth tend to have greater PWG, demonstrating the positive influence of MS score on the postweaning performance of cattle. The direct heritability estimates confirm the possibility of selecting animals for the traits studied.

Evaluating traits that allow breeders to increase production efficiency in beef production systems is important. The mouth size (MS) score is a trait easily measured and implemented by breeders. Our results showed that MS in Nelore cattle is a heritable trait, and it is favorably associated with growth traits, indicating that animals with larger mouth are heavier at weaning and gain more weight after weaning on pasture. MS score should be further explored to evaluate its complexity and inclusion in breeding programs incorporating data collected from cattle raised under pasture conditions.

Genotype-environment interaction for milk production of Gyr cattle in Brazil and Colombia.

BACKGROUND: Genotype by environment interactions (G × E) can play an important role in cattle populations and should be included in breeding programs in order to select the best animals for different environments. OBJECTIVE: The aim of this study was to investigate the G × E for milk production of Gyr cattle in Brazil and Colombia by applying a reaction norm model used genomics information, and to identify genomic regions associated with milk production in the two countries. METHODS: The Brazilian and Colombian database included 464 animals (273 cows and 33 sires from Brazil and 158 cows from Colombia) and 27,505 SNPs. A two-trait animal model was used for milk yield adjusted to 305 days in Brazil and Colombia as a function of country of origin, which included genomic information obtained with a single-step genomic reaction norm model. The GIBBS3F90 and POSTGSf90 programs were used. RESULTS: The results obtained indicate G × E based on the reranking of bulls between Brazil and Colombia, demonstrating environmental differences between the two countries. The findings highlight the importance of considering the environment when choosing breeding animals in order to ensure the adequate performance of their progeny. Within this context, the reranking of bulls and the different SNPs associated with milk production in the two countries suggest that G × E is an important effect that should be included in the genetic evaluation of Dairy Gyr cattle in Brazil and Colombia. CONCLUSION: The Gyr breeding program can be optimized by choosing a selection environment that will allow maximum genetic progress in milk production in different environments within and between countries.

Carcass and meat quality of red-winged tinamou (Rhynchotus rufescens) selected for muscle growth.

The aim of the present study was to evaluate the efficiency of selection for body growth and its association with carcass and meat quality traits in the red-winged tinamou. Two experimental groups were selected based on the selection index: selection group with a higher index (TinamouS) and commercial group with a lower index (TinamouC). Weight at 180 days and slaughter weight were significantly higher (p < 0.05) in the TinamouS group, as were hot carcass weight, skinless breast weight, wing weight, and thigh and drumstick weight. The meat quality traits or sensory attributes did not differ significantly (p > 0.05) between groups. A significant positive correlation (0.59; p < 0.05) was found between shear force and chewability and a significant negative correlation (- 0.59; p < 0.05) between aroma intensity and strange aroma. The present study suggests that the selection index promotes greater body growth and preserves meat quality and sensory traits in red-winged tinamou.

Analysis of residual feed intake in Nellore bulls of different ages, rib eye area, and backfat thickness.

The aim of the present study was to use different models that include body composition phenotypes for the evaluation of residual feed intake (RFI) in Nellore bulls of different ages. Phenotypic and genotypic data of bulls that had participated in feed efficiency tests of a commercial (COM) and an experimental (EXP) herd between 2007 and 2019 were used. The mean entry age in the two herds was 645 and 279 days, respectively. The phenotypes were evaluated: rib eye area (REA), backfat thickness (BFT), residual feed intake (RFIKOCH), RFI adjusted for REA (RFIREA), RFI adjusted for BFT (RFIBFT), and RFI adjusted for REA and BFT (RFIREA BFT). The (co)variance components and prediction of genomic estimated breeding values (GEBV) were obtained by REML using ssGBLUP in single and two-trait analyses. Spearman's correlations were calculated based on the GEBV for RFIKOCH. The RFI phenotypes exhibited moderate heritability estimates in both herds (0.17 ± 0.03 to 0.27 ± 0.04). The genetic correlation between phenotypes was positive and high (0.99) in the two herds, a fact that permitted the creation of a single database (SDB). The heritability estimates of the SDB were also of moderate magnitude for the different definitions of RFI (0.19 ± 0.04 to 0.21 ± 0.04). The genetic correlations were positive and high between RFI traits 0.97 ± 0.01 to 0.99 ± 0.01), and positive and low/moderate between REA and BFT (0.01 ± 0.10 to 0.31 ± 0.12). The selection of animals based on the GEBV for RFIKOCH did not alter the ranking of individuals selected for RFIREA, RFIBFT, and RFIREA BFT. The results of the present study suggest that records of Nellore bulls of different ages and with different body compositions can be combined in a SDB for RFI calculation. Therefore, young animals can be evaluated in feed efficiency tests in order to reduce costs and the generation interval and possibly to obtain a higher response to selection.

Genetic principal components for reproductive and productive traits in Holstein cows reared under tropical conditions.

The objective of this study was to compare the standard multi-trait model and five reduced-rank models fitted to the first principal components and genetic parameter estimates in order to determine the most appropriate method to model the covariance structure of reproductive and productive traits in Brazilian Holstein cows. Individual records of the following traits from 5217 cows were analyzed: 305-day milk yield (MY305), peak yield, milk yield per day of calving interval, days from calving to first estrus, days from calving to last service (CLS), calving interval (CI), and gestation length. Schwarz's Bayesian information criterion was used to compare the different models. The results indicated that four principal components were necessary to model the genetic (co)variance structure, reducing the number of parameters to be estimated. Analysis of genetic and phenotypic correlations showed that milk production-related traits were strongly correlated with each other (ranging from 0.74 to 0.99), while the correlation of these traits with the reproductive traits was weak (ranging from - 0.14 to 0.27). Heritability estimates for the traits ranged from 0.03 to 0.18. The reproductive traits CLS and CI and the production trait MY305 should be included as selection criteria in dairy cattle breeding programs because they are correlated with the first two principal components, retaining 91% of the genetic variability of the data.

Type 2 Marine-Lenhart Syndrome: An Uncommon Cause of Thyrotoxicosis.

Marine-Lenhart syndrome (MLS) is an uncommon cause of primary hyperthyroidism, which can occur in the context of diffuse goiter due to Graves disease (GD) or autonomic nodular disease (Plummer disease (PD)). The coexistence of these two conditions is the hallmark of the MLS. Patients with MLS have a lower remission rate with oral antithyroid drugs, requiring definitive management therapies with radioactive iodine or surgery. We present the case of a 48-year-old female with a history of primary autoimmune hyperthyroidism (GD) since 2016, with biochemical control of hyperthyroidism with methimazole but without the possibility of stopping treatment. The scintigraphic uptake pattern showed heterogeneous uptake of the thyroid parenchyma with three hyper-uptake nodules without inhibition of the rest of the thyroid tissue, findings of an MLS condition with the indication for definitive therapy, for which he was referred to nuclear medicine for the administration of radioactive iodine.

Macrophages and T Lymphocytes in the Ovine Placenta After Experimental Infection With Toxoplasma gondii.

Early abortion in ovine toxoplasmosis has had limited investigation. This study evaluated the immune response in the placenta of sheep orally infected with Toxoplasma gondii and euthanized between 2 and 4 weeks postinfection. Toxoplasma infection of the placenta was only found at 4 weeks after infection. Parasitic debris in foci of necrosis were immunolabeled in the maternal caruncle, whereas well-preserved intracellular parasitic vacuole-like structures were found in trophoblasts of fetal cotyledon. Early abortions had increased macrophages in caruncular septa, whereas in later abortions the placentas containing the parasite had an increase of T lymphocytes and macrophages mainly in the fetal cotyledons. This study suggests that the immune response in both the fetal and maternal compartments of the placenta may contribute to the pathogenesis of ovine toxoplasmosis and that these responses differ between early and late presentations of the disease.

Relationship between reproductive and productive traits in Holstein cattle using multivariate analysis.

Multivariate procedures are used for the extraction of variables from the correlation matrix of phenotypes in order to identify those traits that explain the largest proportion of phenotypic variation and to evaluate the relationship structure between these traits. The reproductive traits (days from calving to first estrus [CFE], days from calving to last service [CLS], calving interval [CI] and gestation length [GL]) and milk production traits (milk yield at 305 days of lactation [MY305], peak yield [PY] and milk yield per day of calving interval [MYCI]) of 5,217 Holstein females (primiparous and multiparous) were measured. Principal component analysis (PCA) and factor analysis of the correlation matrix were used to estimate the correlation between traits. Analysis grouped the seven traits into three principal components and four latent factors that retained approximately 81.5% and 88.9% of the total variation of the data, respectively. The production variables exhibited positive phenotypic correlation coefficients of high magnitude (>.67). The phenotypic correlation estimates between the productive and reproductive traits were low, ranging from .13 to .22. A strong association (.99) was observed between CLS and CI. Our results indicate that multivariate analysis was effective in generating correlations between the traits studied, grouping the seven traits in a smaller number of variables that retained approximately 81% of the total variation of the data.

Positive Culture and Prognosis in Patients With Sepsis: A Prospective Cohort Study.

PURPOSE: To analyze the prognostic role of positive cultures in patients with sepsis. METHODS: A prospective cohort study in a tertiary referral hospital in Medellín, Colombia. Adults older than 18 years of age with a bacterial infection diagnosis according to Centers for Disease Control criteria and sepsis (evidence of organ dysfunction) were included. A logistic regression model was used to determine the association between positive cultures and hospital mortality, and a Cox regression with a competing risk modeling approach was used to determine the association between positive cultures and hospital stay as well as secondary infections. RESULTS: Overall, 408 patients had positive cultures, of which 257 were blood culture, and 153 had negative cultures. Patients with positive cultures had a lower risk of mortality (odds ratio [OR], 0.43; 95% confidence interval [CI], 0.27-0.68), but this association was not maintained after adjusting for confounding factors (OR, 0.56; 95% CI, 0.31-1.01). No association was found with the hospital stay (adjusted subhazard ratio [SHR], 1.06; 95% CI, 0.83-1.35). There was no association between positive cultures and the presence of secondary infections (adjusted SHR, 0.99; 95% CI, 0.58-1.71). CONCLUSION: Positive cultures are not associated with prognosis in patients with sepsis.

Descripción de la experiencia en pacientes diagnosticados con insulinoma. Estudio multicéntrico en Medellín, Colombia / Description of the experience with patients with insulinoma diagnosis. Multicentric study in Medellín, Colombia

RESUMEN Objetivo: describir la experiencia de los pacientes con insulinoma, diagnosticados y tratados entre los años 2002 y 2017 en tres hospitales de la ciudad de Medellín, Colombia. Métodos: estudio descriptivo y retrospectivo de pacientes con criterios bioquímicos para hipoglucemia hiperinsulínica y confirmación histopatológica de insulinoma. Resultados: se estudiaron 19 casos, 10 eran mujeres (52,6 %), la edad media al diagnóstico fue 43 años (D.E 15,5). Hubo cuatro casos de insulinoma multifocal (21,1 %), tres asociados con NEM-1 (15,8 %) y dos malignos (10,6 %). Todos presentaron hipoglucemia en ayunas y 63,2 % posprandial. En la prueba de ayuno, el nadir de glucemia sucedió antes de 48 horas en todos los casos, en promedio 9 horas (D.E 8,0). El diagnóstico bioquímico fue realizado con hipoglucemia e insulina elevada en todos los casos, aunque el péptido C fue reportado en nueve pacientes (47,3 %) y las sulfonilureas en dos (11,1 %). La localización preoperatoria se hizo por imágenes en 12 individuos (68,5 %) y las pruebas invasivas fueron necesarias en seis (31,5 %). Las pruebas diagnósticas fueron positivas en un 83 % para resonancia, 50 % para ecografía endoscópica y prueba de estímulo intraarterial con calcio y 100 % para ecografía intraoperatoria. La cirugía se realizó en 18 casos (94,7 %). La mortalidad (15,8 %) fue derivada de complicaciones en el posoperatorio temprano; la curación se logró en todos los casos. Conclusiones: el insulinoma en nuestro medio tiene características demográficas y clínicas similares a otras series. Existen limitaciones locales para el acceso a los estudios bioquímicos y en el rendimiento diagnóstico de las pruebas de localización.

SUMMARY Objective: The objective of the study was to describe the characteristics of patients with insulinoma in three hospitals in Medellín, Colombia, between 2002 and 2017. Methods: A retrospective analysis of patients with hyperinsulinemic hypoglycemia and histologic confirmation of insulinoma was conducted. Results: A total of 19 cases were identified. Ten women (52.6 %) and 9 males (47.4 %). The mean age at diagnosis was 43 years (S.D: 15.5). Four cases had multifocal insulinoma (21.1%), 3 cases were secondary to multiple endocrine neoplasia type 1 (15.8 %), and 2 of them were malignant (10.6 %). All patients presented fasting hypo-glycemia, and 63.2% presented post-prandial hypoglycemia. Glucose nadir in the fasting test occurred in the first 48 hours in all cases, with a mean time to hypoglycemia of 9 hours (S.D 8.0). The biochemical diagnosis was done with increased insulin in the presence of hypoglycemia. C-peptide was done in 9 patients (47.3 %), and sulfonylureas in 2 cases (11.1 %). Preoperative localization was done by imaging in 12 cases (68.5 %), and invasive tests were required in 6 cases (31.5 %). Localization tests were positive as follows: magnetic resonance imaging in 83%, endoscopic ultrasound in 50%, selective intra-arterial calcium injection in 50 %, and intraoperative ultrasound in 100%. Eighteen patients (94. 7%) underwent surgery. Mortality (15.8 %) was secondary to early post-operative complications. Conclusions: The characteristics of patients with insulinoma in Medellín are similar to other series. However, there are important local limitations for proper biochemical testing and imaging localization. This is the largest study in our country.

Peripheral and placental immune responses in sheep after experimental infection with Toxoplasma gondii at the three terms of gestation.

Although it is known that gestation could influence the clinical course of ovine toxoplasmosis, the precise effect of the term of gestation when sheep are infected are yet mostly unknown. The aim of this study was to evaluate the peripheral and placental immune responses developed in pregnant sheep after experimental infection with Toxoplasma gondii at different times of gestation. Thirty-six pregnant sheep were allocated in different groups, orally inoculated with sporulated oocysts of T. gondii at early, mid and late gestation and culled within 30 days post-infection. The peripheral humoral and cytokine responses were evaluated, as well as the transcription of cytokines at the placenta. Serological analysis revealed that, regardless the term of gestation when infected, specific IgG against T. gondii were detected from day 8 post-infection and there was an early peripheral release of IFN-γ at the first week post-infection followed by a short peak of IL10 and TNF-α at the second week post-infection. There were no significant differences in this response between infected groups. At the placenta, a similar increase in transcription of IFN-γ, and TNF-α was found at the three terms of gestation, while IL-4 increased mainly at the first and second terms and IL-10 transcription was higher at the last term. While these findings show that both Th1 and Th2 cytokines play a key role in the pathogenesis of ovine toxoplasmosis and that placental and peripheral immune responses do not closely correlate, there seems to be no clear modulation of these responses along the gestation.

Antimicrobial agent prescription: a prospective cohort study in patients with sepsis and septic shock.

OBJECTIVE: To assess the true association between appropriate prescription of antibiotics and prognosis in patients with sepsis, a key issue in health care and quality improvement strategies. METHODS: Prospective cohort study in three university hospitals to determine whether the empirical prescription of antibiotics was adequate or inadequate, and to compare hospital death rates and length of stay according to different classifications of antibiotics prescription. Logistic regression models for risk estimation were fitted. RESULTS: A total of 705 patients with severe sepsis were included. No differences were found in positive-culture patients (n = 545) regarding the risk of death with insufficient spectrum antibiotics, compared to patients who received adequate spectrum antibiotics (OR = 0.90; 95% CI = 0.55-1.48). Delay in initiating antibiotics was not associated with the risk of death in patients with adequate spectrum of antibiotics, either with positive (OR = 1.04; 95% CI = 0.99-1.08) or negative cultures (OR = 0.98; 95% CI = 0.92-1.04). There were no differences in the length of hospital stay, according to the antibiotic prescription (median 11 days, IQR = 7-18 days for the whole cohort). CONCLUSIONS: No associations were found between inadequate antibiotic prescription or delay to initiate therapy and mortality or length of stay.

Late diagnosis of pseudohypoparathyroidism in adulthood. Case series / Pseudohipoparatiroidismo diagnosticado tardíamente en la adultez, una serie de casos

Abstract Introduction: Pseudohypoparathyroidism (PHP) is a rare hereditary disease, characterized by hypocalcemia/hyperphosphatemia secondary to peripheral resistance to parathyroid hormone (PTH). PHP diagnosis is usually precluded since hypocalcemia is considered as the primary diagnosis, thus delaying further diagnostic studies and preventing an adequate management of this clinical condition. Materials and methods: Retrospective review of the databases of the Endocrinology departments of two tertiary care centers of Medellin, Colombia from January 2012 to December 2016. Patients diagnosed with PHP based on clinical presentation and confirmatory laboratory values were included. Results: Four patients met the inclusion criteria. All PHP cases were diagnosed in adulthood despite strong early clinical and laboratory evidence of the disease. Three patients were diagnosed with Fahr's syndrome and two with Albright's hereditary osteodystrophy. The mean values obtained were PTH of 376.8 pg/mL, calcium of 6.17 mg/dL and phosphorus of 6.55 mg/dL. Conclusions: PHP is a rare disorder. This paper describes four PHP cases diagnosed during adulthood. Emphasis should be placed on the judicious approach to the patient with hypocalcemia and hyperphosphatemia with increased PTH and normal renal function, since these symptoms strongly suggest a diagnosis of PHP.

Resumen Introducción. El pseudohipoparatiroidismo (PHP) es una condición rara caracterizada por hipocalcemia e hiperfosfatemia secundarias a resistencia periférica a la hormona paratiroidea (PTH). Es frecuente que la hipocalcemia sea establecida de forma equivocada como diagnóstico primario y que el diagnóstico definitivo de PHP sea tardío, difiriendo los estudios y el manejo específico que exigen estos pacientes. Materiales y métodos. Se revisaron de forma retrospectiva las bases de datos de endocrinología de dos centros terciarios de Medellín, Colombia, desde enero de 2012 a diciembre de 2016. Se incluyeron pacientes con diagnóstico de PHP por presentación clínica y valores confirmatorios de laboratorio. Resultados. Cuatro pacientes cumplieron los criterios de inclusión. Todos los casos fueron diagnosticados en la adultez a pesar de tener evidencia temprana, clínica y bioquímica de la enfermedad. Tres pacientes tenían síndrome de Fahr y dos tenían osteodistrofia hereditaria de Albright. Los valores medios registrados fueron PTH de 376.8 pg/mL, calcio de 6.17 mg/dL y fósforo de 6.55 mg/dL. Conclusiones. El PHP es un trastorno raro; se describen cuatro casos diagnosticados de forma tardía en la adultez. Se enfatiza en el enfoque juicioso del paciente con hipocalcemia, la cual, en presencia de hiperfosfatemia con PTH elevada y función renal normal, debe hacer sospechar el diagnóstico de PHP.

Safety and efficacy of the bumped kinase inhibitor BKI-1553 in pregnant sheep experimentally infected with Neospora caninum tachyzoites.

Neospora caninum is one of the main causes of abortion in cattle, and recent studies have highlighted its relevance as an abortifacient in small ruminants. Vaccines or drugs for the control of neosporosis are lacking. Bumped kinase inhibitors (BKIs), which are ATP-competitive inhibitors of calcium dependent protein kinase 1 (CDPK1), were shown to be highly efficacious against several apicomplexan parasites in vitro and in laboratory animal models. We here present the pharmacokinetics, safety and efficacy of BKI-1553 in pregnant ewes and foetuses using a pregnant sheep model of N. caninum infection. BKI-1553 showed exposure in pregnant ewes with trough concentrations of approximately 4 µM, and of 1  µM in foetuses. Subcutaneous BKI-1553 administration increased rectal temperatures shortly after treatment, and resulted in dermal nodules triggering a slight monocytosis after repeated doses at short intervals. BKI-1553 treatment decreased fever in infected pregnant ewes already after two applications, resulted in a 37-50% reduction in foetal mortality, and modulated immune responses; IFNγ levels were increased early after infection and IgG levels were reduced subsequently. N. caninum was abundantly found in placental tissues; however, parasite detection in foetal brain tissue decreased from 94% in the infected/untreated group to 69-71% in the treated groups. In summary, BKI-1553 confers partial protection against abortion in a ruminant experimental model of N. caninum infection during pregnancy. In addition, reduced parasite detection, parasite load and lesions in foetal brains were observed.

Immunohistochemical expression of interferon-γ in different types of granulomatous lesions associated with bovine paratuberculosis.

Animals infected with Mycobacterium avium subsp. paratuberculosis (Map) show a variety of lesions, from focal forms, seen in subclinical stages to diffuse lesions in clinical cases. The purpose of this study was to evaluate the local expression of IFN-γ by immunohistochemistry in relation with the type of lesion in naturally Map-infected cows. The number of immunolabelled cells, -the majority morphologically consistent with lymphocytes-, was higher in focal and diffuse paucibacillary forms than in diffuse multibacillary lesions, where they appeared closely related to epithelioid cells. Diffuse multibacillary lesions had the lowest numbers, but higher than controls, and positive cells were intermingled among the macrophages. The peripheral IFN-γ production was higher in all Map infected cows and a positive correlation was found with the number of immunolabelled cells in the intestine. The findings of this study show that IFN-γ would play a role in the development of the different types of lesions in paratuberculosis, and also points out the importance of adequate sampling of lymphoid tissue containing samples when studying the local immune response in which IFN-γ expression may be involved, especially in cases where focal lesions are present.

Experimental ovine toxoplasmosis: influence of the gestational stage on the clinical course, lesion development and parasite distribution.

The relation between gestational age and foetal death risk in ovine toxoplasmosis is already known, but the mechanisms involved are not yet clear. In order to study how the stage of gestation influences these mechanisms, pregnant sheep of the same age and genetic background were orally dosed with 50 oocysts of Toxoplasma gondii (M4 isolate) at days 40 (G1), 90 (G2) and 120 (G3) of gestation. In each group, four animals were culled on the second, third and fourth week post infection (pi) in order to evaluate parasite load and distribution, and lesions in target organs. Ewes from G1 showed a longer period of hyperthermia than the other groups. Abortions occurred in all groups. While in G2 they were more frequent during the acute phase of the disease, in G3 they mainly occurred after day 20 pi. After challenge, parasite and lesions in the placentas and foetuses were detected from day 19 pi in G3 while in G2 or G1 they were only detected at day 26 pi. However, after initial detection at day 19 pi, parasite burden, measured through RT-PCR, in placenta or foetus of G3 did not increase significantly and, at in the third week pi it was lower than that measured in foetal liver or placenta from G1 to G3 respectively. These results show that the period of gestation clearly influences the parasite multiplication and development of lesions in the placenta and foetus and, as a consequence, the clinical course in ovine toxoplasmosis.

Systemic and local immune responses in sheep after Neospora caninum experimental infection at early, mid and late gestation.

Besides its importance in cattle, Neospora caninum may also pose a high risk as abortifacient for small ruminants. We have recently demonstrated that the outcome of experimental infection of pregnant sheep with 10(6) Nc-Spain7 tachyzoites is strongly dependent on the time of gestation. In the current study, we assessed peripheral and local immune response in those animals. Serological analysis revealed earlier and higher IFN-γ and IgG responses in ewes infected at early (G1) and mid (G2) gestation, when abortion occurred. IL-4 was not detected in sera from any sheep. Inflammatory infiltrates in the placenta mainly consisted of CD8+ and, to a lesser extent, CD4+ T cells and macrophages (CD163+). The infiltrate was more intense in sheep infected at mid-gestation. In the foetal mesenchyme, mostly free tachyzoites were found in animals infected at G1, while those infected in G2 displayed predominantly particulate antigen, and parasitophorous vacuoles were detected in sheep infected at G3. A similar pattern of placental cytokine mRNA expression was found in all groups, displaying a strengthened upregulation of IFN-γ and IL-4 and milder increases of TNF-α and IL-10, reminiscent of a mixed Th1 and Th2 response. IL-12 and IL-6 were only slightly upregulated in G2, and TGF-ß was downregulated in G1 and G2, suggestive of limited T regulatory (Treg) cell activity. No significant expression of TLR2 or TLR4 could be detected. In summary, this study confirms the pivotal role of systemic and local immune responses at different times of gestation during N. caninum infection in sheep.

Immunohistochemical study and mRNA cytokine profile of the local immune response in cattle naturally infected with Calicophoron daubneyi.

In order to recognize the local immune response of the definitive host to Calicophoron daubneyi natural infection, an immunohistochemical study was carried out in the reticulum and rumen in 49 naturally infected cattle. The role of cytokines (IL-4 and IL-10 interleukins and IFN-γ) in the activation of specific defence mechanisms was evaluated by reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) assays to study cytokine mRNA expression. In all infected animals, CD3+ T lymphocytes seemed to be the main element of the inflammatory infiltrate in the reticular and ruminal lamina propria at the point of the parasite adhesion. Intraepithelial globule leukocytes also showed immunolabelling for CD3. Most CD3+ cells also expressed CD4 (T cell helper) antigen although sporadic CD8+-cytotoxic lymphocytes were observed. Local expression of IFN-γ was observed in damaged papillae at the site of parasite attachment and in scattered cells in the lamina propria. B cells (CD79αcy+, CD45+ and IgG+) were found constantly in relation to lymphoid aggregates. MAC387 was expressed in squamous epithelium and in macrophages of the lamina propria of affected papillae. Macrophages in this location also stained positively for CD163 and CD68. Intraepithelial Langerhans cells and macrophages located in the lamina propria showed immunopositivity for MHCII in the affected areas. RT-qPCR analysis confirmed a statistical significant increase of IFN-γ, and IL-10 expression (p<0.01) in the rumen associated with the presence of flukes. These findings suggest a predominant Th1 polarized local immune response with the probable involvement of Th regulatory cells in cattle C. daubneyi natural infection.

Virulence attenuation of a Mycobacterium avium subspecies paratuberculosis S-type strain prepared from intestinal mucosa after bacterial culture. Evaluation in an experimental ovine model.

The differences in pathogenicity between an inoculum derived directly from an intestinal tissue homogenate from a paratuberculosis affected sheep and the S-type Mycobacterium avium subsp. partuberculosis (Map) strain isolated in laboratory media from the mentioned homogenate were assessed in two experiments in lambs. Specific peripheral immune responses were significantly lower in animals inoculated with the cultured organisms that showed only granulomatous lesions in the intestinal lymphoid tissue. However, in the homogenate group, more abundant granulomata also occurred in the lamina propria. Map was isolated only in lambs infected with the culture strain. Map DNA was demonstrated by nested-PCR in all the lambs but in a lower proportion (57.1% vs 100%) in those from the culture group. Under these particular experimental conditions, the results suggest that an attenuation of Map virulence has occurred in the cultured strain compared to the initial tissue homogenate, even after a low number of passages.

Placental thrombosis in acute phase abortions during experimental Toxoplasma gondii infection in sheep.

After oral administration of ewes during mid gestation with 2000 freshly prepared sporulated oocysts of T. gondii isolate M4, abortions occurred between days 7 and 11 in 91.6% of pregnant and infected ewes. Afterwards, a further infection was carried out at late gestation in another group of sheep with 500 sporulated oocysts. Abortions happened again between days 9 and 11 post infection (pi) in 58.3% of the infected ewes. Classically, abortions in natural and experimental ovine toxoplasmosis usually occur one month after infection. Few experimental studies have reported the so-called acute phase abortions as early as 7 to 14 days after oral inoculation of oocysts, and pyrexia was proposed to be responsible for abortion, although the underline mechanism was not elucidated. In the present study, all placentas analysed from ewes suffering acute phase abortions showed infarcts and thrombosis in the caruncullar villi of the placentomes and ischemic lesions (periventricular leukomalacia) in the brain of some foetuses. The parasite was identified by PCR in samples from some placentomes of only one sheep, and no antigen was detected by immunohistochemical labelling. These findings suggest that the vascular lesions found in the placenta, and the consequent hypoxic damage to the foetus, could be associated to the occurrence of acute phase abortions. Although the pathogenesis of these lesions remains to be determined, the infectious dose or virulence of the isolate may play a role in their development.