Protected biofactors and antioxidants reduce the negative consequences of virus and cold challenge while enhancing performance by modulating immunometabolism through cytoskeletal and immune signaling in the jejunum.

The aim of this study was to evaluate the effectiveness and mechanism of action of 2 feed additives in reducing the impacts of virus and temperature stressors. We determined the effects of protected biofactors and antioxidants (P(BF+AOx)), and protected biofactors and antioxidants with protected organic acids and essential oils (P(BF+AOx)+P(OA+EO)) on the immune and metabolic health of Ross 308 broiler chickens. These biofactors and antioxidants were derived from vitamins, and Aspergillus niger, Aspergillus oryzae and Bacillus subtilis fermentation extracts. All Ross 308 chickens were exposed to a double-dose of live bronchitis vaccine at d 0 and environmentally challenged by reducing the temperature from 32°C to 20°C at d 3 for 48 h. Control birds were fed without feed additives in the diet. Performance data and jejunum samples were collected to evaluate the effects of these treatments on growth, cytokine expression, and protein phosphorylation via kinome peptide array. ANOVA was used for statistical analysis of the performance and gene expression data (p-value of 0.05), and PIIKA2 was used for statistical evaluation and comparison of the kinome peptide array data. The P(BF plus;AOx) and P(BF+AOx)+P(OA+EO) treatments significantly increased bird weight gain and decreased feed conversion. The kinome peptide array data analysis showed increased activity of cytoskeletal, cell growth and proliferation proteins, and metabolic signaling in the jejunum of P(BF+AOx)+P(OA+EO) treated chickens. There was a significant decrease in IL-6 gene expression in the jejunum of P(BF+AOx)+P(OA+EO) samples compared to control at d 15. P(BF+AOx)+P(OA+EO) treatments in the jejunum showed strong immunomodulatory effects, perhaps to control inflammation. P(BF+AOx)+P(OA+EO) improves gut health via growth and metabolic signaling in the jejunum while inducing stronger immunomodulation.

A protected complex of biofactors and antioxidants improved growth performance and modulated the immunometabolic phenotype of broiler chickens undergoing early life stress.

We evaluated the supplementation of a protected complex of biofactors and antioxidants [P(BF+AOx)] on growth performance, antioxidant activity, expression of immune-related genes, and immunometabolic phenotype of broilers submitted to early life stressors. The treatments were a nutritionally complete basal diet supplemented or not with P(BF+AOx) (Jefo Nutrition Inc., Saint-Hyacinthe, QC, Canada) from 1 to 14 d of age. 720 one-day old male Ross 308 chickens were placed into pens of 30 birds (12 replicates/treatment). Birds were double-vaccinated against infectious bronchitis (IB; MILDVAC-Ma5T) at the hatchery and submitted, on d 3, to an acute reduction on environmental temperature (from 32° C to 20°C) for 48 h. Feed intake (FI), body weight gain (BWG), and feed conversion ratio (FCR) were calculated weekly. On d 7 and 15, samples were collected for expression of immune-related genes and kinome array analysis, and serum to evaluate the antioxidant status. Data were analyzed by ANOVA using SAS (SAS 9.4). From d 1 to 21 and d 1 to 28, the dietary supplementation of P(BF+AOx) significantly increased BWG (P < 0.05) by 3.6 and 3.8%, respectively, and improved FCR (P < 0.05) by 1.2 and 1.8%, respectively. From d 1 to 35, dietary supplementation enhanced BWG (P = 0.03) by 4%. Serum glutathione reductase activity on d 15 was higher in birds fed diets supplemented with P(BF+AOx) compared to the control diet-fed birds (P = 0.04). Dietary supplementation reduced the expression of IL-1ß (P = 0.03) in the lungs on d 7. On d 15, dietary supplementation increased the expression of IL-6 (P = 0.02) and IL-10 (P = 0.03) in the liver. It was observed that, via decreased phosphorylation, catalase was activated in the jejunum and liver, and the phosphorylation of immunoregulatory or proinflammatory proteins was decreased. Other important cellular signaling pathways were also changed in the liver and jejunum due to the supplementation. The supplementation of P(BF+AOx) improves growth performance by promoting a general anti-inflammatory and antioxidant response in chickens undergoing early life stress.

Mycotoxin-contaminated diets and an adsorbent affect the performance of Nellore bulls finished in feedlots.

Mycotoxins are present in almost all feedstuffs used in animal nutrition but are often ignored in beef cattle systems, even though they can affect animal performance. The objective of this study was to evaluate the effects of mycotoxins and a mycotoxin adsorbent (ADS) on performance of Nellore cattle finished in a feedlot. One hundred Nellore cattle (430 ± 13 kg) were used in a randomized complete block design with a 2 × 2 factorial arrangement of treatments. The factors consisted of two diets with either natural contamination (NC) or exogenous contamination (EC) and the presence (1 g/kg of DM; ADS) or absence of a mycotoxin adsorbent. The NC and EC diets had the following contaminations, respectively: 0.00 and 10.0 µg/kg aflatoxins, 5114 and 5754 µg/kg fumonisins, 0.00 and 42.1 µg/kg trichothecenes B, 0.00 and 22.1 µg/kg trichothecenes A and 42.9 and 42.9 µg/kg fusaric acid. At the beginning of the experiment, all animals were weighed, and four randomly selected animals were slaughtered to evaluate the initial carcass weight. After 97 days of treatment, all animals were weighed and slaughtered. There was no interaction among factors for the DM intake (DMI; P = 0.92); however, there was a tendency for the EC diets to decrease the DMI by 650 g/day compared to animals fed NC diets (P = 0.09). There was a trend for interaction among factors (P = 0.08) for the average daily gain (ADG), where the greatest ADG was observed for cattle fed the NC diet (1.77 kg), and the lowest was observed for those fed the EC diet (1.51 kg). The NC + ADS and EC + ADS treatments presented intermediate values for ADG. The animals fed the NC diet had a greater final BW (596 kg) than animals fed the EC treatment (582 kg; P = 0.04). There was a tendency for interaction among factors for carcass gain (P = 0.08). Similarly to ADG, the highest carcass gain was observed for animals fed the NC diet (1.20 kg), and the lowest was observed for those fed the EC diet (1.05 kg). The NC + ADS and EC + ADS treatments presented intermediate values. The natural contamination groups had greater carcass gain than that of the EC groups, and the use of the ADS recovered part of the weight gain in animals fed the EC diet. In conclusion, mycotoxins at the levels evaluated affected the performance of beef cattle, and adsorbents may mitigate their impact.

Effect of functional oils on the immune response of broilers challenged with Eimeria spp.

Infection with Eimeria sp. results in the activation of multiple facets of the host immune system; the use of phytogenics can modulate the inflammatory response and improve the performance of the challenged animal. The aim of this study was to evaluate the effect of a commercial blend of cashew nut shell liquid (CNSL) and castor oil on the immune response of broilers challenged with coccidiosis. A total of 864 one-day-old male chicks (Cobb 500) were randomly distributed into six treatment groups (8 pens/treatment and 18 chicks/pen) in a three-by-two factorial design with three additives: control (non-additive), 100 ppm of monensin or 0.15% CNSL-castor oil. Challenge status was determined twice at 14 days of age. Unchallenged birds were inoculated by gavage with oocysts sporulated with Eimeria tenella, Eimeria acervulina and Eimeria maxima. Although the positive control (non-additive and challenged) and CNSL-castor oil treatment groups exhibited similar variation in weight gain (ΔBWG) compared to unchallenged birds fed without additives, the variation observed in birds fed diets containing CNSL-castor oil was associated with a higher maintenance requirement and not feed efficiency. In the second week after infection, ΔBWG of the CNSL-castor oil treatment group did not significantly change compared to the other treatment groups. At days 7 and 14 post-challenge, there was a higher excretion of oocysts in the control group, whereas the CNSL-castor oil and monensin groups did not differ. The CNSL-castor oil group exhibited increased gene expression of interferon (IFN), interleukin 6 (IL-6) and tumor necrosis factor (TNF), while the control group exhibited increased expression of cyclooxygenase (COX) and IL-1. The heterophils/lymphocyte ratio was low for the monensin treatment group. The unchallenged birds that received monensin treatment presented higher gene expression of IFN, COX and IL-1 compared to the other treatments, while the CNSL-castor oil group exhibited reduced gene expression, except for TNF. The commercial blend of cashew nut liquid and castor oil modulated the inflammatory response against Eimeria spp. In the absence of the parasite, there was no stimulation of genes involved in the inflammatory response, demonstrating that the blend is an effective tool in specifically modulating the immune system of birds afflicted with coccidiosis.

Histological parameters to evaluate intestinal health on broilers challenged with Eimeria and Clostridium perfringens with or without enramycin as growth promoter.

The maintenance of integrity of the gastrointestinal tract is an important aspect for animal productivity, since it is able to absorb nutrients more efficiently and serves as a barrier against microorganisms. To control agents detrimental to intestinal integrity, growth-promoting antibiotics (AGP) are used, which reduce the number of toxin-producing microorganisms in the intestinal lumen, acting as anti-inflammatory agents. There is a demand for restriction of use of AGP in animal feed, but there are few studies showing what parameters we should observe to search for alternative additives. The aim of this study was to establish histological parameters that explain the effect of enramycin as growth promoter on intestinal health in broilers challenged with Eimeria and Clostridium perfringens. The zootechnical performance and the histology by I See Inside (ISI) methodology were evaluated on liver and ileum samples. Chickens challenged without AGP have the worst BWG, FCR, and histological ISI score (ISI score 9) in the ileum compared to non-challenged (ISI score 5). The use of enramycin on challenged group significantly minimized the ISI score in the ileum at 21 and 28 d (ISI score 7.4 and 8.0, respectively) compared with the challenged group not fed with enramycin (ISI score 9.2 and 9.9, respectively), associated with reduced lamina propria thickness and inflammatory cell infiltration. We suggest these 2 histological parameters as a standard to compare products for gut health.

Effect of aflatoxin experimental ingestion and Eimeira vaccine challenges on intestinal histopathology and immune cellular dynamic of broilers: applying an Intestinal Health Index.

The present study evaluated the effect of aflatoxin B1 and Eimeria vaccine challenges and their interaction on intestinal morphology, applying the morphometric index "I See Inside" (ISI). Immune cellular response and broiler chicken performance were also studied. A total of 240 broiler chickens were divided in a 2 × 2 factorial arrangement with 4 treatments, T1: Control diet and no challenge (CON), T2: Aflatoxin B1 (AFLA), T3: Control diet and Eimeria challenge (COC), and T4: Aflatoxin B1 and Eimeria challenge (AFLA+COC). The mathematical morphometric index ISI was applied to evaluate macro and microscopic alterations. Samples of liver and jejunum were analyzed for macrophages, CD4+, and CD8+ cells counting by immunohistochemistry at 7, 14, and 21 d of age. Chickens challenged with Eimeria presented higher ISI of macroscopic alterations associated to Eimeria lesion at the medium small intestine, lower body weight gain (BWG) and feed intake (FI), and worse feed conversion ratio compared to non-challenged birds. Both Eimeria and aflatoxin challenges modulated the immune cells in the jejunum and liver, generally increasing the number of macrophages, CD4+, and CD8+ cells in relation to the control group. Birds from COC and COC+AFLA groups presented higher ISI histological score in the jejunum at 7 and 14 d of age compared to the CON and AFLA groups. The reduction of FI and BWG was correlated to high histological ISI and resulted in a high presence of immune cells in tissues, suggesting immune response demand. The histological ISI had statistical correlation to broiler performance.

Effects of conjugated linoleic acid and lutein on the growth performance and immune response of broiler chickens.

The effects of lutein and conjugated linoleic acid (CLA) on growth performance and immune response of broiler chickens were evaluated in the presence and absence of Salmonella lipopolysaccharide (LPS) immune challenge. Cobb chicks (360; 1 to 22 d of age) were used in a 3 × 2 factorial arrangement of CLA (0, 1, and 2%) and lutein (0 and 50 mg/kg) dietary levels. At d 8 and 15, birds were injected with BSA to assess IgY production. At d 20, birds were injected with LPS. Samples of liver, spleen, and duodenum were collected at 3 and 16 h post-LPS challenge for RT-qPCR analysis of RXRα, RXRγ, PPARα, PPARγ, TLR-4, IL-1ß, IL-2, IL-10, and IL-12 gene expression. CLA decreased BW, BW gain (BWG), and G:F from d 1 to 20, but these effects were reversed when lutein was included in the 1% CLA diet (P < 0.001). The production of IgY anti-BSA increased following a 2% CLA supplementation (P < 0.01). LPS increased the liver:BW ratio at 3 h post-injection (P < 0.001) and decreased BWG at 3, 16, and 40 h (P < 0.001). Lutein decreased plasmatic nitric oxide levels (P < 0.01). LPS downregulated PPARα mRNA in the duodenum (P = 0.02) and liver (P = 0.04), and PPARγ (P = 0.01) and RXRα (P = 0.08) in the spleen; these effects were not reversed by CLA or lutein as initially hypothesized. Although LPS upregulated IL-1ß (P = 0.02) and IL-12 (P = 0.07) expression, lutein downregulated these pro-inflammatory cytokines in the liver (P = 0.03 and P = 0.07, respectively). Lutein decreased splenic (P = 0.09) but increased hepatic (P = 0.06) TLR-4 mRNA. A dietary CLA supplementation of 2% increased hepatic RXRα (P = 0.10). In conclusion, CLA decreased broiler chicken growth performance, but lutein could prevent this negative effect (depending on CLA dose). Lutein had an anti-inflammatory effect, and a 2% CLA supplementation improved the humoral immune response.

Inoculation of newly hatched broiler chicks with two Brazilian isolates of Salmonella Heidelberg strains with different virulence gene profiles, antimicrobial resistance, and pulsed field gel electrophoresis patterns to intestinal changes evaluation.

Salmonella Heidelberg is one of the 3 most frequently isolated serovars from human Salmonella cases in Canada, and the fourth most commonly reported Salmonella serovar in human foodborne disease cases in the United States. Since 1962, Salmonella Heidelberg has been isolated and reported in poultry and poultry products in Brazil. The poultry industry has focused efforts on reducing salmonellae incidence in live production in an effort to reduce Salmonella in the processing plant. A better understanding of the initial infection in chicks could provide approaches to control Salmonella contamination. The objective of the present study was to evaluate 2 Salmonella Heidelberg strains that differed in the presence of virulence genes invA, agfA, and lpfA; antimicrobial resistance profiles; and epidemiologic profiles on aspects of pathogenicity and intestinal morphology. Newly hatched broiler chicks were inoculated with 2 strains (SH23 and SH35) of Salmonella Heidelberg and cecal morphometry, histopathology, electron microscopy, and bacterial counts in the liver and cecum were assessed. The SH23 and SH35 strains resulted in different changes in villi height and crypt depth and inflammatory cell infiltration in the cecum. The SH35 group had higher liver and cecum bacterial cell counts when compared with SH23 strains.

Characterization of a spontaneously transformed chicken mononuclear cell line.

We describe the characterization of a spontaneously transformed chicken monocytic cell line that developed as a single colony of cells in a heterophil culture that was inadvertently left in the incubator over a period of 25 days. These cells, hitherto named HTC, grow efficiently at both 37 or 41 degrees C in culture medium containing either 5% FBS or 2% chicken serum. The HTC cells are acid phosphatase positive, show expressions of both class I and class II major histocompatibility complex (MHC), CD44, K1, and K55 cell surface antigens, and engulf latex beads, produce nitrite and interleukin-6 on stimulation with bacterial lipopolysaccharide (LPS). Treatment with phorbol myristate acetate (PMA) induces respiratory burst in HTC cells and the secretion of matrix metalloproteinase (MMP) into culture medium. Using gene-specific primers and reverse transcriptase-polymerase chain reaction (RT-PCR), the presence of mRNA trancripts for interferon-gamma (IFN-gamma), interleukin-1 (IL-1), interleukin-6 (IL-6), nitric oxide synthase (NOS), matrix metalloproteinase-2 (MMP-2), and transforming growth factor-beta (TGF-beta) were detected. Lipopolysaccharide (LPS) treatment of HTC cells modulated IL-1, IL-6, IFN-gamma, NOS mRNA levels as detected by RT-PCR analyses. Using different avian tumor virus gene-specific primers and PCR, the HTC cells were positive for the presence of avian leukosis virus (ALV) and Marek's disease virus (MDV) but negative for reticuloendothelial virus (REV), chicken infectious anemia virus (CIAV), and herpes virus of turkeys (HVT). The production of ALV antigens by HTC cells was further confirmed using p27 gag protein ELISA. Collectively, these results show that the HTC cells belong to myeloid/macrophage lineage and were likely transformed by ALV and MDV but retain many interesting and useful biological activities.

Influência da infusäo transcervical de plasma seminal ou de estrógeno na concepçäo, no ciclo estral e na ovulaçäo de porcas / Influence of transcervical infusion of seminal plasma or oestrogen on conception, oestrus cycle and ovulation in sows

O objetivo deste trabalho foi o de avaliar o desempenho de 211 porcas da linhagem Camborough 22 Marca Registrada, de diferentes ordens de parto, submetidas à infusäo transcervical de plasma seminal ou de estrógeno no início do estro. Após o desmame, elas foram examinadas duas vezes ao dia para a detecçäo do estro, na presença de um macho sexualmente maduro, e da ovulaçäo, com auxílio da ultrasonografia transcutânea. No início do estro as fêmeas receberam aleatoriamente uma infusäo transcervical de plasma seminal, de estrógeno ou permaneceram como grupo-controle. Todas as fêmeas foram inseminadas no turno seguinte à detecçäo do estro, recebendo no máximo três inseminaçöes com intervalos de 8 a 16h. No grupo-controle as fêmeas de primeiro parto ovularam mais precocemente que as fêmeas com maior número de partos (P<0,05). Os tratamentos näo influenciaram a duraçäo do estro e o momento da ovulaçäo. O número total de leitöes nascidos näo diferiu entre os tratamentos

Influência da suplementaçäo de glutamina sobre o desempenho e o desenvolvimento de vilos e criptas do intestino delgado de frangos / Influence of glutamine supplementation on performance and intestinal villous and crypt development in broiler chickens

Investigou-se o efeito da suplementaçäo de glutamina na dieta sobre o consumo de raçäo, ganho de peso e conversäo alimentar e sobre a estrutura da mucosa intestinal de frangos. Foram utilizados 320 pintos de corte machos distribuídos em um delineamento inteiramente ao acaso com dois tratamentos e quatro repetiçöes, sendo T1 suplementado com 1 por cento de L-glutamina na dieta e T2 controle. Os índices de desempenho foram analisados aos 7, 21 e 49 dias de idade das aves. Aos 7 e 14 dias de idade oito aves foram sacrificadas para colheita de fragmentos de cada porçäo do intestino delgado para avaliaçäo da morfometria intestinal em microscopia de luz em sistema analisador de imagens "Video Plan". As variáveis estudadas foram altura dos vilos, profundidade de cripta e relaçäo vilo: cripta. A adiçäo de 1 por cento de glutamina à dieta de frangos näo influenciou (P>0,05) o seu desempenho zootécnico. Entretanto, 1 por cento de glutamina na raçäo foi capaz de alterar (P<0,01) a altura do vilo, a profundidade de cripta e a relaçäo vilo:cripta no duodeno, bem como a altura de vilo do íleo de frangos no sétimo dia de idade

Ca(2+)-independent nitric oxide synthase activity in human lung after cardiopulmonary bypass.

BACKGROUND: Because surgery involving cardiopulmonary bypass induces a systemic inflammatory response, the effect of cardiopulmonary bypass on nitric oxide (NO) generation was investigated in human lung tissue. METHODS: Nitric oxide synthase (NOS) activity was measured by the conversion of 14C-L-arginine to 14C-L-citrulline in tissue biopsy samples obtained before and after cardiopulmonary bypass. RESULTS: The Ca(2+)-independent production of NO found before cardiopulmonary bypass was extremely low (1.5 (0.5) pmol citrulline/mg/min), but was increased after the bypass operation (23.6 (11) pmol/mg/min). CONCLUSIONS: Ca(2+)-independent NOS activity was detected in human lung after cardiopulmonary bypass. This finding may provide an important insight into the pathogenesis of the tissue damage and acute phase response observed after such surgery.