RESUMO
Necrotic enteritis (NE) is a disease of worldwide distribution, which affects young broilers and causes economic losses on a scale of 6 billion dollars per year. For decades, NE was controlled in poultry flocks by dietary administration of low doses of antimicrobial growth promoters (AGPs). However, an increase in NE incidence was noted after the AGP ban. This study aimed to compare the effect of an antibiotic (Enramycin) diet to a combination of sodium butyrate, hydrolyzed yeast, and zinc proteinate (ViligenTM) on broiler diets regarding performance, blood parameters, intestinal permeability, morphology and lesions, and carcass yield of broilers challenged with Eimeria spp. and Clostridium perfringens to simulate subclinical necrotic enteritis. A total of 1,150 one-day-old male broiler chickens with an initial average weight of 43.9 ± 0.65 g were allocated to 50 experimental pens. Animals were divided into 5 groups: Negative control (NC) without additives; Positive control (PC) with 0.12 g/ton of Enramycin (8%); V500, V1000, and V1500 with the addition of 500, 1.000, and 1.500 g/ton of Viligen, respectively. All animals were challenged by Eimeria spp. at 7 d of age and by C. perfringens at 17, 18, and 19 d for induction of subclinical NE. The broilers fed with all concentrations of Viligen showed similar performance, blood parameters, intestinal permeability, and carcass yield compared to PC broilers. However, NC broilers showed higher FCR compared to PC broilers from 1 to 33 d (1.42 vs. 1.39) (P = 0.048) and from 1 to 42 d (1.51 vs. 1.49) (P < 0.001). V1500 broilers had fewer intestinal lesions at 28 d when compared to the PC treatment (P < 0.05) and showed that higher Viligen inclusion resulted in lower intestinal damage. At 21 d, the V500 group showed higher intestinal morphology characteristics (VH:VD 4.9 vs. 3.5) compared to the PC treatment (P < 0.001). Thus, in this study, the dietary addition of Viligen to broilers challenged by an experimental model of subclinical NE resulted in lower intestinal damage and similar performance to that obtained by the addition of Enramycin.
Assuntos
Ração Animal , Galinhas , Infecções por Clostridium , Clostridium perfringens , Coccidiose , Dieta , Eimeria , Enterite , Doenças das Aves Domésticas , Animais , Galinhas/crescimento & desenvolvimento , Doenças das Aves Domésticas/parasitologia , Doenças das Aves Domésticas/microbiologia , Ração Animal/análise , Masculino , Dieta/veterinária , Enterite/veterinária , Enterite/parasitologia , Coccidiose/veterinária , Coccidiose/parasitologia , Infecções por Clostridium/veterinária , Eimeria/fisiologia , Clostridium perfringens/fisiologia , Suplementos Nutricionais/análise , Antibacterianos/administração & dosagem , Antibacterianos/farmacologia , Ácido Butírico/administração & dosagem , Distribuição Aleatória , Necrose/veterinária , Peptídeos CíclicosRESUMO
OBJECTIVES: This study aimed to produce and purify Clostridium perfringens type C beta-toxin, sheep anti-beta toxin immunoglobulin G (IgG) and chicken immunoglobulin Y (IgY). METHODS: Two methods were used for beta-toxin purification: single-step metal affinity chromatography (MAC) using zinc as a chelator and ion exchange chromatography (IEX). The purified and inactivated beta-toxoids were then administered to sheep and chickens in order to produce IgG and IgY. RESULTS: All assays using the IEX failed. In contrast, MAC purified more than 21 mg of toxin per run in a single-step protocol. The purified and inactivated beta-toxoids were then administered to sheep and chickens, and IgG and IgY were purified with a high yield, medium antibody titer of 50 IU/mL, and high avidity (73.2 %). CONCLUSIONS: C. perfringens type C beta-toxin and sheep or chicken anti-beta toxin IgG and IgY antibodies were successfully produced and purified using a simple protocol. This protocol can be used for the production of components used in the diagnosis and research of necrotic enteritis caused by C. perfringens type C, as well as for the evaluation of existing vaccines and the development of new preventive methods against this disease.
Assuntos
Antitoxinas , Infecções por Clostridium , Enterite , Imunoglobulinas , Doenças das Aves Domésticas , Animais , Ovinos , Clostridium perfringens , Infecções por Clostridium/veterinária , Enterite/veterinária , Galinhas , Toxoides , Imunoglobulina G , Doenças das Aves Domésticas/prevenção & controleRESUMO
Deoxynivalenol (DON) is a predisposing factor for necrotic enteritis. This study aimed to investigate the effects of a DON and Clostridium perfringens (CP) challenge on the intestinal morphology, morphometry, oxidative stress, and immune response of broilers. Additionally, we evaluated the potential of a Lactobacillus spp. mixture as an approach to mitigate the damage induced by the challenge. One-day-old broiler chickens (n = 252) were divided into seven treatment groups: Control, DON, CP, CP + DON, VL (DON + CP + viable Lactobacillus spp. mixture), HIL (DON + CP + heat-inactivated Lactobacillus spp. mixture), and LCS (DON + CP + Lactobacillus spp. mixture culture supernatant). Macroscopic evaluation of the intestines revealed that the CP + DON group exhibited the highest lesion score, while the VL and HIL groups showed the lowest scores. Microscopically, all Lactobacillus spp. treatments mitigated the morphological changes induced by the challenge. DON increased levels of reactive oxygen species (ROS) in the jejunum, and CP increased ROS levels in the jejunum and ileum. Notably, the Lactobacillus spp. treatments did not improve the antioxidant defense against CP-induced oxidative stress. In summary, a Lactobacillus spp. mixture, whether used as a probiotic, paraprobiotic, or postbiotic, exerted a partially protective effect in mitigating most of the intestinal damage induced by DON and CP challenges.
Assuntos
Galinhas , Probióticos , Tricotecenos , Animais , Clostridium perfringens , Espécies Reativas de Oxigênio , Intestinos , Lactobacillus , Probióticos/farmacologiaRESUMO
This study performed the characterization of coccidiosis in broilers and evaluated the occurrence of suggestive cases of necrotic enteritis (NE), seeking if there is an association between the diseases in Brazilian flocks. Two hundred and fifty-six birds from 32 flocks were evaluated. Macroscopic and histopathological lesions were graduated for coccidiosis and NE. Intestinal content was investigated by polymerase chain reaction (PCR) for seven species of Eimeria and by selective anaerobic culture for Clostridium perfringens and identification of the NetB gene. Flocks positive for coccidiosis represented 93.8%. Macroscopic lesions of coccidiosis were Grade 1 for E. acervulina (27%); E. tenella (9.7%) and E. maxima (8.9%). Histopathological evaluation showed Grade 1 in duodenum (38.2%); jejunum (21.4%); cecum (9.3%) and ileum (5%). PCR demonstrated positivity for E. tenella (21.9%), E. maxima (18.8%), and E. acervulina (3.1%). Suggestive macroscopic lesions of necrotic enteritis ranged from Grade 1 (16%), 2 (23%) and 3 (10,9%). Histopathology indicated the absence of necrosis, showing only hemorrhage in the mucosa and submucosa, with the presence of Eimeria spp. Clostridium perfringens type A netB+ was not isolated, demonstrating that macroscopic lesions found mostly in the jejunum did not characterize NE, based on histopathology and negativity of the NetB gene. The study suggests that, due to the high occurrence of coccidiosis, many macroscopic findings suggestive of NE are, in fact, attributed to atypical lesions caused by the reproduction of Eimeria spp.
Este estudo realizou a caracterização de coccidiose em frangos de corte e avaliou a ocorrência de casos sugestivos de enterite necrótica (EN), buscando se há alguma associação entre estas duas enfermidades em lotes de frango de corte no Brasil. Foram avaliadas 256 aves de 32 lotes. Lesões macroscópicas e histopatológicas foram graduadas para coccidiose e EN. O conteúdo intestinal foi investigado por reação em cadeia da polimerase (PCR) para sete espécies de Eimeria e por cultura anaeróbia seletiva para Clostridium perfringens e identificação do gene NetB. Os lotes positivos para coccidiose representaram 93,8%. Lesões macroscópicas de coccidiose foram de Grau 1 para E. acervulina (27%); E. tenella (9,7%) e E. maxima (8,9%). A avaliação histopatológica mostrou Grau 1 no duodeno (38,2%); jejuno (21,4%); ceco (9,3%) e íleo (5%). A PCR demonstrou positividade para E. tenella (21,9%), E. maxima (18,8%) e E. acervulina (3,1%). Lesões macroscópicas sugestivas de enterite necrótica variaram de grau 1 (16%), 2 (23%) e 3 (10,9%). A histopatologia indicou ausência de necrose, apresentando apenas hemorragia em mucosa e submucosa, com presença de Eimeria spp. Clostridium perfringens tipo A netB + não foi isolado, demonstrando que lesões macroscópicas encontradas principalmente no jejuno não caracterizaram NE, com base na histopatologia e negatividade do gene NetB. O estudo sugere que, em virtude da alta ocorrência de coccidiose nos lotes, muitos achados macroscópicos sugestivos de EN são, na verdade, atribuídos a lesões atípicas provocadas pela reprodução de Eimeria spp.
Assuntos
Animais , Feminino , Galinhas/microbiologia , Coccidiose/patologia , Coccidiose/veterinária , Coccidiose/epidemiologia , Enterite/diagnóstico , Enterite/veterinária , Enterite/patologiaRESUMO
Clostridium perfringens is the etiological agent of NE, a disease that greatly affects the poultry industry. Experiments on the induction of NE are difficult to carry out, as it is a multifactorial disease, and thus different predisposing factors have been used. This study evaluated the effect of the Gumboro disease vaccine virus vaccine (IBDV-vac) associated or not with infection by Eimeria spp. in broilers, as a predisposing factor for NE. Broilers (n = 99) were divided into groups (11) challenged with IBDV-vac, Eimeria spp. CP type G (CP13, CP14 and CP03) or both. The macroscopic evaluation revealed that the highest average (3.45) of injury occurred for the CP13 + IBDV-vac group. The microscopic analysis showed that Eimeria spp. increased the population of intraepithelial lymphocytes and reduced the villus/crypt ratio in duodenum and jejunum when associated with CP13 or CP14. There was a synergistic effect between the CP strain used and the predisposing factors; nevertheless, it was not clear which was the most effective predisposing factor to potentiate the lesions, suggesting that the association of the strain with the factors should first be evaluated for each experimental protocol.
RESUMO
The use of antibiotic growth promoters (AGPs) has historically been the most important prophylactic strategy for the control of Necrotic Enteritis (NE) caused by some Clostridium perfringens toxin types in poultry. During the last five decades, AGPs have also been supplemented in feed to improve body weight gain and feed efficiency as well as to modulate the microbiome (consisting of microbes and their genes both beneficial and potentially harmful) and reduce enteric pathogens, among other benefits. New regulatory requirements and consumer preferences have led to strong interest in natural alternatives to the AGPs for the prevention and control of illnesses caused by enteric pathogens. This interest is not just focused on the direct removal or inhibition of the causative microorganisms but also the improvement of intestinal health and homeostasis using a range of feed additives. A group of promising feed additives is short- and medium-chain fatty acids (SCFA and MCFA, respectively) and their derivatives. The use of SCFA and MCFA, including butyric, caproic, caprylic, capric, and lauric acids, has shown strong effects against NE in broilers both at experimental and commercial levels. These fatty acids also benefit intestinal health integrity and homeostasis. Other effects have also been documented, including increases in intestinal angiogenesis and gene expression of tight junctions. Chemical modifications to improve stability and point of release in the intestine have been shown to improve the efficacy of SCFA and MCFA and their derivatives. The aim of this review is to give an overview of SCFA, MCFA and their derivatives, as an alternative to replace AGPs to control the incidence and severity of NE in poultry.
RESUMO
Clostridium perfringens (Cp.) is the cause of human foodborne desease. Meat and poultry products are identified as the main source of infection for humans. Cp. can be found in poultry litter, feces, soil, dust, and healthy birds' intestinal contents. Cp. strains are known to secrete over 20 identified toxins and enzymes that could potentially be the principal virulence factors, capable of degrading mucin, affecting enterocytes, and the small intestine epithelium, involved in necrotic enteritis (NE) pathophysiology, also leading to immunological responses, microbiota modification and anatomical changes. Different environmental and dietary factors can determine the colonization of this microorganism. It has been observed that the incidence of Cp-associated to NE in broilers has increased in countries that have stopped using antibiotic growth promoters. Since the banning of such antibiotic growth promoters, several strategies for Cp. control have been proposed, including dietary modifications, probiotics, prebiotics, synbiotics, phytogenics, organic acids, and vaccines. However, there are aspects of the pathology that still need to be clarified to establish better actions to control and prevention. This paper reviews the current knowledge about Cp. as foodborne pathogen, the pathophysiology of NE, and recent findings on potential strategies for its control.
RESUMO
Decreases in the use of antibiotics and anticoccidials in the poultry industry have risen the appearance of necrotic enteritis (NE). The purpose of this study was to evaluate the effect of a Bacillus direct-fed microbial (DFM) on growth performance, intestinal integrity, NE lesions and ileal microbiota using a previously established NE-challenged model. At day-of-hatch, chicks were randomly assigned to three different groups: Negative control (NC), Positive control (PC) challenged with Salmonella Typhimurium (day 1), Eimeria maxima (EM, day 13) and Clostridium perfringens (CP, day 18-19), and Bacillus-DFM group (DFM) challenged as the PC. Body weight (BW) and body weight gain (BWG) were measured weekly. Total feed intake (FI) and feed conversion ratio (FCR) were evaluated at day 21. Liver samples were collected to assess bacterial translocation and blood samples were used to measure superoxide dismutase (SOD) and fluorescein isothiocyanate-dextran (FITC-d). Intestinal contents were obtained for determination of total IgA and microbiota analysis. NE lesion scores (LS) were performed at day 21. Chickens consuming the DFM significantly improved BW and had a numerically more efficient FCR compared to PC at day 21. Additionally, there were no significant differences in FCR between the DFM group and NC. Furthermore, the DFM group showed significant reductions in LS, IgA and FITC-d levels compared to the PC. However, there were no significant differences in SOD between the groups. The microbiota analysis indicated that the phylum Proteobacteria was significantly reduced in the DFM group in comparison to PC. At the genus level, Clostridium, Turicibacter, Enterococcus, and Streptococcus were reduced, whereas, Lactobacillus and Bacillus were increased in the DFM group as compared to PC (p < 0.05). Likewise, the DFM significantly reduced CP as compared to PC. In contrary, no significant differences were observed in bacterial composition between NC vs. DFM. In addition, beta diversity showed significant differences in the microbial community structure between NC vs. PC, and PC vs. DFM. These results suggest that the dietary inclusion of a selected DFM could mitigate the complex negative impacts caused by NE possibly through mechanism(s) that might involve modulation of the gut microbiota.
RESUMO
Two experiments were conducted to evaluate the effect of the prophylactic or therapeutic administration of a 0.1% mixture containing ascorbic acid and a solid dispersion of curcumin with polyvinylpyrrolidone and boric acid (AA-CUR/PVP-BA) against Salmonella Enteritidis (S. Enteritidis) in broiler chickens. A third experiment was conducted to evaluate the impact of the dietary administration of 0.1% AA-CUR/PVP-BA in a necrotic enteritis (NE) model in broiler chickens. The prophylactic administration of 0.1% AA-CUR/PVP-BA significantly decreased S. Enteritidis colonization in cecal tonsils (CT) when compared to the positive control group (PC, p < 0.05). The therapeutic administration of 0.1% AA-CUR/PVP-BA significantly reduced the concentration of S. Enteritidis by 2.05 and 2.71 log in crop and CT, respectively, when compared with the PC on day 10 post-S. Enteritidis challenge. Furthermore, the serum FITC-d concentration and total intestinal IgA levels were also significantly lower in chickens that received 0.1% AA-CUR/PVP-BA. Contrary, the PC group showed significantly higher total intestinal IgA levels compared to the negative control or AA-CUR/PVP-BA groups in the NE model. However, 0.1% AA-CUR/PVP-BA showed a better effect in reducing the concentration of S. Enteritidis when compared to the NE model. Further studies with higher concentration of AA-CUR/PVP-BA into the feed to extend these preliminary results are currently being evaluated.
RESUMO
A study was conducted to investigate the effects of Bacillus amyloliquefaciens CECT 5940 as a direct-fed microbial (DFM) alone or in association with bacitracin methylene disalicylate (BMD) in broilers under enteric pathogen challenge. A total of 1,530-day-old male Cobb500 chicks were randomly assigned to 5 treatments, with 9 replicate pens with 34 birds each. Treatments included positive control (PC, basal diet without additives or challenge); negative control (NC, basal diet without additive and challenged birds); NC + 0.05 g/kg BMD; NC + 1 g/kg DFM (106 CFU B. amyloliquefaciens CECT 5940/g of feed); and NC + 0.05 g/kg BMD + 1 g/kg DFM. The challenge consisted of oral gavage with Eimeria maxima and Clostridium perfringens inoculum. Body weight gain (BWG), feed intake (FI), and feed conversion ratio (FCR) were evaluated on days 21, 35, and 42. Ileal and cecal content were collected on days 21 and 28 for C. perfringens enumeration by real-time PCR assay and the intestinal health was evaluated by scores. Uniformity (UN), carcass (CY), and breast meat yields (BMY) were evaluated on day 42. After 14 and 21 d post-inoculation, birds in the challenged groups had significant lower FI and BWG compared to the PC group (P < 0.05). However, the groups receiving DFM, BMD, or its combination presented better FCR, CY, BMY, UN, and lower incidence of footpad lesion and litter quality visual scores, compared to the NC group without feed additives (P < 0.05). Mortality was not affected by treatments (P > 0.05). Broilers fed DFM, BMD, or its combination presented lower C. perfringens in ileal content at 21 and 28 d compared to NC group without additives (P < 0.05) and also maintained gut health by keeping the frequency of ballooning, abnormal content, and swollen mucosa comparable to the PC group (P > 0.05). The study indicates that Bacillus amyloliquefaciens CECT 5940 is effective as BMD to provide similar performance and gut health in challenged broilers.