Liver transcriptomic networks reveal main biological processes associated with feed efficiency in beef cattle.

BACKGROUND: The selection of beef cattle for feed efficiency (FE) traits is very important not only for productive and economic efficiency but also for reduced environmental impact of livestock. Considering that FE is multifactorial and expensive to measure, the aim of this study was to identify biological functions and regulatory genes associated with this phenotype. RESULTS: Eight genes were differentially expressed between high and low feed efficient animals (HFE and LFE, respectively). Co-expression analyses identified 34 gene modules of which 4 were strongly associated with FE traits. They were mainly enriched for inflammatory response or inflammation-related terms. We also identified 463 differentially co-expressed genes which were functionally enriched for immune response and lipid metabolism. A total of 8 key regulators of gene expression profiles affecting FE were found. The LFE animals had higher feed intake and increased subcutaneous and visceral fat deposition. In addition, LFE animals showed higher levels of serum cholesterol and liver injury biomarker GGT. Histopathology of the liver showed higher percentage of periportal inflammation with mononuclear infiltrate. CONCLUSION: Liver transcriptomic network analysis coupled with other results demonstrated that LFE animals present altered lipid metabolism and increased hepatic periportal lesions associated with an inflammatory response composed mainly by mononuclear cells. We are now focusing to identify the causes of increased liver lesions in LFE animals.

Effects of ß-Glucan Supplementation on LPS-Induced Endotoxemia in Horses.

ß-glucan is part of the cell wall of fungi and yeasts and has been known for decades to have immunomodulating effects on boosting immunity against various infections as a pathogen-associated molecular pattern that is able to modify biological responses. ß-glucan has been used in rat models and in vitro studies involving sepsis and SIRS with good results, but this supplement has not been evaluated in the treatment of endotoxemia in horses. This study aims to evaluate the effects of preventive supplementation with ß-glucan in horses submitted to endotoxemia by means of inflammatory response modulation. Eight healthy horses, both male and female, aged 18 ± 3 months, weighing 300 ± 100 kg of mixed breed, were randomly assigned to two groups of four animals, both of which were subjected to the induction of endotoxemia via the intravenous administration of E. coli lipopolysaccharides (0.1 µg/kg). For 30 days before the induction of endotoxemia, horses in the ß-glucan group (GB) received 10 mg/kg/day of ß-glucan orally, and horses in the control group (GC) received 10 mg/kg/day of 0.9% sodium chloride orally. The horses were submitted to physical exams, including a hematological, serum biochemistry, and peritoneal fluid evaluation, and the serum quantification of cytokines TNF-α, IL-6, IL-8, and IL-10. For statistical analysis, the normality of residues and homogeneity of variances were verified; then, the variables were analyzed as repeated measures over time, checking the effect of treatment, time, and the interaction between time and treatment. Finally, the averages were compared using Tukey's test at a significance level of 5%. Horses from both experimental groups presented clinical signs and hematological changes in endotoxemia, including an increase in heart rate and body temperature, neutrophilic leukopenia, an increase in serum bilirubin, glucose, lactate, and an increase in TNF-α, IL-6, and IL-10. Hepatic and renal function were not compromised by ß-glucan supplementation. GB presented higher mean values of the serum total protein, globulins, and IL-8 compared to that observed in GC. In the peritoneal fluid, horses from GB presented a lower mean concentration of neutrophils and a higher mean concentration of macrophages compared to the GC. It was concluded that preventive supplementation of ß-glucan for thirty days modulated the immune response, as evidenced by increasing serum total proteins, globulins, IL-8, and changes in the type of peritoneal inflammatory cells, without effectively attenuating clinical signs of endotoxemia in horses. Considering the safety of ß-glucan in this study, the results suggest the potential clinical implication of ß-glucan for prophylactic use in horse endotoxemia.

Complications following transcutaneous cecal trocarization in horses with a cattle trocar and a cecal needle.

Percutaneous decompression of the cecum is a procedure that could be considered for horses with cecal gas distension. The aim of this study was to identify complications such as peritonitis and clinically relevant peritonitis (CRP) after transabdominal cecal trocarization in healthy horses using a cattle trocar and a cecal needle. Mixed breed horses were assigned to three groups (n = 6): horses that underwent trocarization with a cecal needle (G1) or a cattle trocar (G2), and a control group (CG) without cecal trocarization. The same horses were used in each group, respecting a three-month washout period between studies. A physical examination, serial blood, and peritoneal fluid sampling were performed, prior to cecal trocarization and 2, 6 and 12 hours after the first collection and 1, 2, 3, 7, and 14 days after the procedure. Acute-phase proteins in blood and peritoneal fluid were analyzed by polyacrylamide gel electrophoresis. Horses with a high cell count in the peritoneal fluid (i.e., 10,000 cells/µl) were considered to have peritonitis and CRP if they met at least two of the following clinical criteria: anorexia, lethargy, tachycardia, tachypnea, fever, ileus, abnormal oral mucous membrane color, abnormal white blood cells count, or high blood fibrinogen concentration (> 5 g/L). All horses recovered from cecal trocarization and abdominocentesis with no major complications. Cecal trocarization caused cytologic evidence of peritonitis in G1 and G2 during the 14 days of evaluation. CRP was not observed, although a decrease in cecal motility was observed in G1 and G2 during the experimental period and three horses, one from G1 and two from G2, showed a single moment of fever. None of the groups showed leukopenia or leukocytosis, although blood neutrophil count decreased at D7 and D14 in G1 and at D14 in G2 (p ≤ 0.05). After cecal trocarization, an increase in the total nucleated cells count, total proteins, globulins, alkaline phosphatase and acute phase proteins were observed in the peritoneal fluid of G1 and G2 during the 14 days of evaluation (p ≤ 0.05), without causing clinically relevant peritonitis. Transcutaneous cecal trocarization promotes peritonitis, which is more intense with a cattle trocar than with a cecal needle. The cecal needle should be considered for cecal trocarization of horses with cecal tympany.

Nephrin and Podocin mRNA Detection in Urine Sediment of Dogs with Chronic Kidney Disease: Preliminary Observations.

Introduction: Dogs with chronic kidney disease (CKD) may have alterations in the glomerular filtration barrier, including podocyte loss. Detection of podocyte mRNA in urine could be useful for assessing podocyturia in dogs with kidney disease. The objective of this study was to evaluate the presence of nephrin mRNA (NPHS1) and podocin mRNA (NPHS2) in urine sediments of dogs with naturally occurring CKD and healthy dogs. Material and Methods: Twenty-four dogs, 14 with CKD and 10 as healthy controls, underwent clinical evaluation. The dogs with CKD were divided into two groups, according to the International Renal Interest Society criteria: stage 1 or 2 CKD (n = 5) and stage 3 or 4 CKD (n = 9). Urine was collected by catheterisation or free catch and RNA isolation from the urine sediments was optimised using glycogen as a co-precipitant. Detection of NPHS1 and NPHS2 in the sediment samples was performed using quantitative real-time PCR. Results: Both types of mRNA were detected in samples from all groups, but the percentages of detection were higher in the group of dogs with stage 1 or 2 CKD and lower in the group of dogs with stage 3 or 4 disease. Conclusion: Physiological podocyturia was observed in healthy dogs, and the results suggest differential podocyturia in dogs with CKD, according to the stage of the disease, i.e. an increase in podocyturia in dogs at stage 1 or 2 and a reduction in podocyturia in dogs at stage 3 or 4.

Congenital Transmission of Toxoplasma gondii After Experimental Reinfection With Brazilian Typical Strains in Chronically Infected Sheep.

Toxoplasma gondii is a cause of congenital diseases, miscarriages and stillbirths in production animals. In Brazil, non-archetypal genotypes of the parasite may be related to severe disease. Experimental infection with T. gondii was studied in sheep to analyse congenital transmission-related parameters in reinfections with different Brazilian parasite strains. Thirteen T. gondii-seronegative sheep were orally infected with 2 × 103oocysts for the primary infection: G1 (4 animals) were inoculated with TgCatBr71 strain (Type BrI genotype) and G2 andG3 (5 and 4 animals, respectively) withTgCatBr60 strain (Type BrIII genotype). After chronification of infection, the animals were impregnated. A second infection was performed after 60 days of gestation. TheG1 andG3 animals were inoculated withTgCatBr60BrIII and the G2 animals withTgCatBr71 BrI oocysts. The effects of reinfection were compared with a control group (5 animals) through physical examination, ultrasound imaging and serology. Ovine experimental infections were evaluated using mouse bioassays, molecular analysis, serological tests, histopathology, and immunohistochemistry. No abortions occurred; a seropositive lamb and a mummified fetus from G2-BrIIIxBrI were produced. The vertical transmission rate detected in lambs from chronically infected sheep was 31.6% (6/19). It is demonstrated that reinfection and subsequent congenital transmission occured in one sheep with a primary Brl infection challenged with BrIII genotype of T. gondii. In a twin pregnancy from G2-BrIIIxBrI, congenital transmission from a latent infection was detected in both lambs. Congenital transmission could not be tracked in three lambs. Overall, previous T. gondii infection may fail to protect against congenital transmission from a reinfection and primary infection induced insufficient protection against vertical transmission which must be taken into account in decision-making for the use of seropositive animals as breeders. Similar trials with larger groups and contemplating host cellular immune response studies should be conducted to evaluate the actual impact of T. gondii reinfection involving different strains in sheep.

Determination of serum aflatoxin B1-lysine to evaluate the efficacy of an aflatoxin-adsorbing feed additive in pigs fed an aflatoxin B1-contaminated diet.

In this study, serum aflatoxin B1 (AFB1)-lysine was determined in order to evaluate the in vivo efficacy of a hydrated sodium calcium aluminosilicate (HSCAS) in pigs fed AFB1. Twenty-four 49-day-old crossbred barrows were maintained in individual cages and allowed ad libitum access to feed and water. A completely randomized design was used with six animals assigned to each of four dietary treatments for 21 days as follows: (A) basal diet (BD), (B) BD supplemented with 0.5 % HSCAS, (C) BD supplemented with 1.1 mg/kg AFB1, and (D) BD supplemented with 0.5 % HSCAS and 1.1 mg/kg AFB1. HSCAS was able to alleviate the toxic effects of AFB1 on pigs and reduce (P < 0.05) the levels of serum AFB1-lysine. Cumulative reductions of adduct yield values, calculated through the equation [(pg AFB1-lysine/mg albumin) / (µg AFB1/kg body weight)], were 53.0, 62.8, and 72.1 after 7, 14, and 21 days of oral exposure, respectively. AFB1-lysine has potential as an AFB1-specific biomarker for diagnostic purposes and for evaluating the efficacy of chemoprotective interventions in pigs.

Evaluation of coagulation parameters in dogs with overweight or obesity

Background: Hemostatic alterations have already been well characterized in humans with body fat excess, being includedin the list of obesity related comorbidities. Overweight and obesity are common in dogs; however, there is little informationabout the blood coagulation parameters in dogs with these conditions. The aim of this study was to compare hematologicaland coagulation parameters between lean dogs and overweight or obese dogs, including platelets count; prothrombin time(PT); activated partial thromboplastin time (aPTT), coagulation time (CT) and plasma fibrinogen concentration.Materials, Methods & Results: A total of 22 dogs (aged 1 to 10 years, neutered) were evaluated, 10 of them presentedideal body condition score (BCS 4-5) and formed the group 1 (control) and 12 were overweight or obese (BCS 7-9) andformed the group 2. The dogs were submitted to clinical evaluation and then to blood collection for the following laboratorytests: blood count (performed on automatic analyzer), quantification...(AU)

Evaluation of coagulation parameters in dogs with overweight or obesity

Background: Hemostatic alterations have already been well characterized in humans with body fat excess, being includedin the list of obesity related comorbidities. Overweight and obesity are common in dogs; however, there is little informationabout the blood coagulation parameters in dogs with these conditions. The aim of this study was to compare hematologicaland coagulation parameters between lean dogs and overweight or obese dogs, including platelets count; prothrombin time(PT); activated partial thromboplastin time (aPTT), coagulation time (CT) and plasma fibrinogen concentration.Materials, Methods & Results: A total of 22 dogs (aged 1 to 10 years, neutered) were evaluated, 10 of them presentedideal body condition score (BCS 4-5) and formed the group 1 (control) and 12 were overweight or obese (BCS 7-9) andformed the group 2. The dogs were submitted to clinical evaluation and then to blood collection for the following laboratorytests: blood count (performed on automatic analyzer), quantification...

Resposta proliferativa de células mononucleares do sangue periférico frente a membranas eritrocitárias autólogas de cães recentemente imunizados / Proliferative response of peripheral blood mononuclear cells against autologous red blood cell membranes of dogs recently immunized

Embora faltem evidências diretas da relação causal entre a vacinação recente e o desenvolvimento da anemia hemolítica imunomediada (AHIM) em cães, pode ser identificada uma associação temporal entre elas. Constituem-se em objetivos deste trabalho avaliar: a presença de imunoglobulinas (IgG e IgM) e complemento (C3>) na superfície eritrocitária e o potencial do estímulo mitogênico de membranas eritrocitárias autólogas sobre os linfócitos periféricos de cães nos momentos pré-vacinal (imediatamente antes da vacinação com vacinas polivalente e antirrábica) e pós-vacinal (28 a 38 dias após a imunização). Vinte e um cães adultos e hígidos, machos e fêmeas, foram submetidos à anamnese, exame físico e avaliações laboratoriais nos dois momentos do estudo. O teste da antiglobulina direta (n=15) foi realizado com o reagente de Coombs polivalente, nas diluições de 1:2 a 1:8. A detecção de imunoglobulinas (IgG, IgM) e complemento (C3) na superfície de eritrócitos por citometria de fluxo (n=21) foi realizada utilizando anticorpos anti-IgG de cão produzido em ovelha cadeia pesada, anti-IgM produzido em cabra e anti-C3 de cão produzido em cabra, todos conjugados com fluoresceína de isotiocianato (FITC). As células mononucleares do sangue periférico foram isoladas por gradiende, marcadas com CFSE e estimuladas com Concanavalina A (ConA) e com membranas eritrocitárias autólogas em duas concentrações (ME1 e ME2). Foi utilizado o Índice de Proliferação (IP) como indicador da proliferação celular, obtido pela divisão das intensidades de fluorescência obtidas por citometria de fluxo das amostras basal e estimulada. As comparações das variáveis \"hemácias marcadas com anti-Ig/C3\" e \"IP de linfócitos\" foram realizadas utilizando-se o t-Student para amostras pareadas. As comparações dos IP de linfócitos frente aos diferentes antígenos (Con A, ME1 e ME2) foram realizadas por meio da ANOVA com medidas repetidas. Quando houve diferença significante entre os índices, foram realizadas comparações múltiplas (teste de Bonferroni). Foi considerado um nível de significância de 5%. Observou-se que os cães se encontravam em boas condições de saúde, nos dois momentos do estudo, com as variáveis hematológicas e bioquímicas mantidas próximas entre si e resultado do teste da antiglobulina direta negativo (n=21). A porcentagem de hemácias marcadas com IgG e IgM nos momentos pré-vacinal (1,06±0,49% e 1,42±1,59%) e pós-vacinal (0,83±0,56% e 1,35±1,71%) não foi alterada, com p=0,261 e p=0,699, respectivamente. A porcentagem de hemácias com C3 na superfície no momento pós-vacinação (0,40±0,38%) foi, em média, menor do que no momento pré-vacinação (0,71±0,33%), com p=0,019. Os índices de proliferação obtidos com a ConA, ME1 e ME2 no momento pré-vacinal (2,15±0,83; 1,03±0,07; 1,05±0,11) e pós- vacinal (2,13±0,58; 1,02±0,05; 1,02±0,05) não se modificaram, com p=0,935; p=0,845 e p=0,222, respectivamente. Em ambos os momentos, os índices de proliferação celular observados com o uso de ConA foram, em media, maiores do que os índices com ME1 e ME2 (p<0,001). A baixa porcentagem de hemácias com IgG, IgM ou C3 na superfície e a ausência de resposta proliferativa dos linfócitos quando estimulados com membranas eritrocitárias, indicam que, neste experimento, não houve nenhuma evidência de que o estímulo vacinal pudesse estar relacionado ao desenvolvimento da AHIM

Despite the lack of evidence regarding a causal link between recent vaccination and development of immune mediated hemolytic anemia (IMHA) in dogs, a temporal association between them has been identified in some cases. The aim of this study was to evaluate: the presence of immunoglobulins (IgG and IgM) and complement (C3) on the surface of red blood cells and the potential for mitogenic stimulation of peripheral lymphocytes against autologous red blood cell membranes on dogs at pre-vaccination (immediately prior to vaccination with polyvalent and anti-rabies vaccines) and post-vaccination (after 28 and 38 days after vaccination). Twenty-one healthy adult dogs (both males and females) were subjected to physical examination and complementary laboratory exams in the aforementioned two instances of the study (i.e. pre-vaccination and post-vaccination). Direct antiglobulin test (n=15) was performed using the polyvalent Coombs reagent in 1:2 to 1:8 dilutions. Immunoglobulins (IgG, IgM) detection and identification of complement (C3) on the surface of red blood cells were done by flow cytometry (n=21) using antibodies anti-dog IgG heavy chain produced in sheep, anti-dog IgM produced in goat and anti-dog C3 produced in goat - all in conjunction with fluorescein isothiocyanate (FITC). The peripheral blood mononuclear cells were isolated by gradient, labeled with CFSE and stimulated with concanavalin A (Con A) and autologous erythrocyte membranes in two concentrations (EM1 and EM2). The Proliferation Index (PI), used as an indicator of cell proliferation, was obtained by dividing the fluorescence intensities of basal and stimulated samples, both obtained by flow cytometry. A comparison was made between the variables \"labeled red blood cells with anti-Ig/C3\" and \"PI lymphocyte\" using the paired Student\'s t-test. Comparisons of PI lymphocytes to different antigens (Con A, EM1 and EM2) were performed using ANOVA of repeated measures. Whenever significant differences between the indices were found, multiple comparisons (Bonferroni test) were then performed. A 5% significance level was considered. At the two instances of the study, dogs were presented in good health status with both hematological and biochemical variables kept close together and negative results for direct antiglobulin test (n=21). The percentage of red blood cells labeled with IgG and IgM in the pre-vaccination (1.06±0.49% and 1.42±1.59%) and post-vaccination (0.83±0.56% and 1.35±1.71%) were kept similar, with p=0.261 and p=0.699, respectively. The percentage of erythrocytes with C3 on the surface at the time post-vaccination (0.40±0.38%) was on average lower than in the pre-vaccination (0.71±0.33%), p=0.019. The proliferation index obtained with ConA, ME1 and ME2 in the pre-vaccination (2.15±0.83, 1.03±0.07, 1.05±0.11) and post-vaccination (2.13±0.58, 1.02±0.05, 1.02±0.05) did not differ significantly, with p=0.935, p=0.845 and p=0.222, respectively. In both instances, rates of cell proliferation observed with the use of ConA were, on average, higher than the rates with ME1 and ME2 (p<0.001). The low percentage of erythrocytes with IgG, IgM or C3 on the surface and the absence of lymphocyte proliferative response when stimulated with erythrocyte membranes, indicate that, at least in this experiment, there is no evidence of any association between vaccine stimulation and development of IMHA in dogs