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1.
Poult Sci ; 101(8): 101988, 2022 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-35809347

RESUMO

Prevention of necrotic enteritis (NE), caused by Clostridium perfringens (C. perfringens), is one of the most important goals to improve the profitability of broiler chickens. This work aimed to compare the efficacy of 2 antibiotic alternatives including a postbiotic (dry feed additive and aqueous nonviable Lactobacillus (L.) species fermentation) and a probiotic (dry feed additive and aqueous Bacillus (B.) subtilis and B. lischeniformis mixture) with an antibiotic (amoxicillin in water) against NE. Four hundred, day-old broiler chicks were divided into 8 equal groups (Gs), n = 50 each (5 replicates; 10 each). Chickens of G1 (postbiotic dry-feed additive), G2 (postbiotic and antibiotic in drinking water), G3 (postbiotic dry and aqueous), G4 (probiotic dry-feed additive), G5 (probiotic and antibiotic in drinking water), G6 (probiotic dry and aqueous), and G7 (nontreated) were orally inoculated with a toxigenic C. perfringens type A on the d 19 to 21 of age and predisposed with 3X coccidial vaccine for induction of NE. However, chickens of G8 were kept nontreated or challenged. The severity of NE signs was markedly decreased in G3 in comparison with other challenged treatment groups, and the mortality rates were 22%, 10%, 16%, 22%, 12%, 20%, and 36% in Gs 1, 2, 3, 4, 5, 6, and 7, respectively. The best significant (P ≤ 0.05) feed conversion ratio was detected in G3 (1.51), G6 (1.54), and G2 and G8 (1.61). In addition, the European production efficiency factor was significantly (P ≤ 0.05) improved in G3 (279.33) and G2 (266.67), but it was decreased in G7 (177.33) when compared with G8 (339.33). An improvement in intestinal and hepatic pathology and liver function tests, as well as a significant (P ≤ 0.05) decrease in bacterial counts were observed in Gs 2, 5, 3, 6, 1, and 4, respectively in comparison with G7. Immunologically, the highest significant (P ≤ 0.05) hemagglutination inhibition antibody titers for Newcastle disease virus vaccine were in Gs 1 and 3 (6.4 log2). In conclusion, the combined feed and water postbiotic treatment demonstrated promising results in ameliorating the severity of NE and improving the hepatic and the immune status of broiler chickens when compared with the commonly used probiotic and antibiotic.


Assuntos
Infecções por Clostridium , Água Potável , Enterite , Doenças das Aves Domésticas , Probióticos , Animais , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Galinhas/fisiologia , Infecções por Clostridium/microbiologia , Infecções por Clostridium/prevenção & controle , Infecções por Clostridium/veterinária , Clostridium perfringens/fisiologia , Enterite/microbiologia , Enterite/prevenção & controle , Enterite/veterinária , Doenças das Aves Domésticas/tratamento farmacológico , Doenças das Aves Domésticas/microbiologia , Doenças das Aves Domésticas/prevenção & controle , Probióticos/farmacologia , Probióticos/uso terapêutico
2.
BMC Infect Dis ; 22(1): 617, 2022 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-35840943

RESUMO

BACKGROUND: Liposuction has become one of the most popular cosmetic surgeries in China. However, few studies have discussed infectious shock caused by C. perfringens as one of the causes of death after liposuction. CASE PRESENTATION: A 24-year-old woman was brought to the emergency department (ED) of Guangzhou Chinese Overseas Hospital for treatment. The patient had undergone liposuction in her bilateral lower limbs two days prior. At the ED, the patient was unconscious, and had bilateral equal-sized (diameter, 6 mm) round pupils, no light reflex, a blood pressure (BP) of 71/33 mmHg, a heart rate of 133 bpm, and an SpO2 of 70%. She had bilateral limb swelling, extensive ecchymoses in her lower abdomen and bilateral thighs, local crepitus, blisters, weak pulses on her femoral artery and dorsalis pedis, high skin tension, and hemoglobin of 32 g/L. The patient was diagnosed with Clostridium perfringens infection, and she underwent debridement surgery and supportive treatment. But the patient's BP could not improve. At 8:28 pm on the day of admission, the patient was declared clinically dead after the electrocardiograph showed a horizontal line and spontaneous respiration ceased. CONCLUSIONS: Failure to meet surgical disinfection and environmental standards may be the cause of infection of C. perfringens through wounds. Therefore, it is necessary to strengthen the environmental disinfection of the operating room, and standardize the sterile conditions of the operation staff and patients before and during operation. Liposuction surgery necrotizing fasciitis is a rare but fatal complications, especially if diagnosis delay, therefore it is critical for early diagnosis and treatment of gas gangrene.


Assuntos
Infecções por Clostridium , Fasciite Necrosante , Gangrena Gasosa , Lipectomia , Choque , Adulto , Infecções por Clostridium/diagnóstico , Clostridium perfringens , Fasciite Necrosante/diagnóstico , Fasciite Necrosante/etiologia , Fasciite Necrosante/terapia , Feminino , Gangrena Gasosa/diagnóstico , Humanos , Lipectomia/efeitos adversos , Choque/complicações , Adulto Jovem
3.
Vet Res ; 53(1): 59, 2022 Jul 26.
Artigo em Inglês | MEDLINE | ID: mdl-35883183

RESUMO

Arginine (Arg), lysine (Lys), and methionine (Met) can be used to support the health status of turkeys. The present study investigated selected performance, gut integrity, and immunological parameters in turkeys reared in optimal or challenge conditions. The experiment lasted for 28 days, and it had a completely randomized 2 × 3 factorial design with two levels of dietary Arg, Lys and Met (high or low) and challenge with Clostridium perfringens (C. perfringens), Escherichia coli lipopolysaccharide (LPS) or no challenge (placebo). Increased dietary levels of Arg, Lys and Met had a beneficial effect on turkey performance and immunological parameters, and it improved selected indicators responsible for maintaining gut integrity in different challenge conditions. Under optimal conditions (with no challenge), high ArgLysMet diets did not compromise bird performance and they improved selected performance parameters in challenged birds. The immune system of turkeys was not excessively stimulated by high ArgLysMet diets, which did not disrupt the redox balance and had no negative effect on gut integrity. High ArgLysMet diets increased the expression levels of selected genes encoding nutrient transporters and tight junction proteins. However, the influence exerted by different dietary inclusion levels of Arg, Lys and Met on gut integrity was largely determined by the stressor (C. perfringens vs. LPS). Further studies are required to investigate the role of Arg, Lys and Met levels in the diet on the immune response, gut function and performance of turkeys in different challenge conditions.


Assuntos
Lisina , Perus , Ração Animal/análise , Animais , Arginina/metabolismo , Clostridium perfringens , Dieta/veterinária , Suplementos Nutricionais , Lipopolissacarídeos , Metionina , Perus/metabolismo
4.
Mol Immunol ; 148: 45-53, 2022 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-35665660

RESUMO

Phospholipases C (PLCs) represent an important group of lethal toxins produced by pathogenic bacteria of the Clostridium genus, including the beta toxin of C. haemolyticum. Bacillary hemoglobinuria in cattle and sheep is the main disease caused by this pathogen and its incidence can be reduced by annual vaccination of herds. Currently, widely used vaccines depend on cultivating the pathogen and obtaining high concentrations of the toxin, disadvantages that can be overcome with the use of recombinant vaccines. In the development of this new generation of immunizing agents, identifying and understanding the structural and immunological aspects of the antigen are crucial steps, but despite this, the beta toxin is poorly characterized. Fortunately, the time and resources required for these investigations can be reduced using immunoinformatics. To advance the development of recombinant vaccines, in addition to a brief review of the structural and immunological aspects of beta toxin, this work provides in silico mapping of immunodominant regions to guide future vaccinology studies against C. haemolyticum. A review of alternatives to overcome the limitations of beta toxin vaccines (conventional or recombinant) is also proposed.


Assuntos
Toxinas Bacterianas , Infecções por Clostridium , Animais , Vacinas Bacterianas , Bovinos , Clostridium , Infecções por Clostridium/prevenção & controle , Clostridium perfringens , Proteínas Recombinantes , Ovinos , Fosfolipases Tipo C , Vacinas Sintéticas
5.
Braz J Biol ; 84: e258114, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35649037

RESUMO

The study was aimed to analyse the effects of antibiotic growth promoters (AGPs), Oxytetracycline di-hydrate and Tylosin phosphate on the intestinal microflora in broiler chicken. The AGPs were provided in different concentrations solely or in combinations for 42 days of rearing. Faecal samples were collected from the intestine (duodenum, jejunum and caeca) of broiler chicken on 14th, 28th and 42nd days of trial. Samples were cultured on different selective medium and bacterial identification was performed by different biochemical and molecular diagnostic tools. Results showed a significant effect of AGPs on the growth of pathogenic microorganisms such as Escherichia coli and Clostridium perfringens in the intestine. Interestingly, an impaired growth was observed for both bacterium showing a significant effect (P<0.05) of AGPs on E. coli and C. perfringens on day 14th, 28th, and 42nd. This effect was observed solely and in combination while using AGPs. Data further showed that the effect was more prominent in combination and with an increase concentration of AGPs. Remarkably, no impairment was seen on the growth of L. reuteri at different sites of intestine and duration (14th, 28th, and 42nd days). The results showed that the use of AGPs in diet has no harmful effect on beneficial bacteria, however, an impaired growth was seen on the harmful bacteria. It is suggested that a combination of AGPs (OXY-1.0+TP-0.5) is economical and have no harmful effect on the broiler chicken. The use of AGPs in a recommended dose and for a specific period of time are safe to use in poultry both as growth promoter and for the prevention of diseases.


Assuntos
Microbioma Gastrointestinal , Oxitetraciclina , Animais , Antibacterianos/farmacologia , Bactérias , Galinhas/microbiologia , Clostridium perfringens , Escherichia coli , Oxitetraciclina/farmacologia , Fosfatos , Tilosina/farmacologia
6.
Arch Microbiol ; 204(7): 361, 2022 Jun 04.
Artigo em Inglês | MEDLINE | ID: mdl-35662380

RESUMO

This is a culture-dependent study with the objective of pure culturing and characterizing pathogenic bacteria from the blowhole, lung, stomach and fecal samples of a neonatal crucially endangered Yangtze finless porpoise (Neophocaena asiaeorientalis asiaeorientalis) that died 27 days after birth. Bacteria were inoculated using a swab onto blood and MacConkey agar plates and representative isolates were identified through 16S rRNA gene sequence analysis. A total of three Clostridium perfringens type C strains from the fecal samples were isolated. Toxin genes, including cpa, cpb and cpb2, were detected by PCR amplification, whereas the etx, iap and cpe genes were not detected. Biofilm formation of the three strains was then examined. Only one strain was capable of biofilm formation. In addition, isolates showed strong resistance against the antibiotics amikacin (3/3), erythromycin (1/3), gentamicin (3/3), streptomycin (3/3), and trimethoprim (3/3), while sensitivity to ampicillin (3/3), bacitracin (3/3), erythromycin (2/3), penicillin G (3/3), and tetracycline (3/3). The results suggested C. perfringens type C could have contributed to the death of this neonatal porpoise.


Assuntos
Toninhas , Animais , Antibacterianos/farmacologia , Bactérias/genética , Biofilmes , Clostridium perfringens/genética , Eritromicina , Genótipo , Toninhas/genética , Toninhas/microbiologia , RNA Ribossômico 16S/genética
7.
Methods Mol Biol ; 2521: 173-188, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35732998

RESUMO

Bacterial toxins gain growing attention as potential cancer treatment due to their potent cytotoxic effects. Among the very different toxins with diverse modes of action, the Clostridium perfringens enterotoxin (CPE) is in focus to treat solid cancers. This toxin targets the tight junction proteins claudin-3 and -4 (Cldn-3/4), which are frequently overexpressed in solid cancers. Binding to these claudins induces pore formation in the host cell plasma membrane leading to rapid oncoleaking cell death of tumor cells. Based on this, extending the targeting of CPE beyond Cldn-3/4 is of interest, since other claudins, such as claudin-1 or -5 are often overexpressed in various cancer entities such as non-small-cell lung cancer (NSCLC) or papillary thyroid carcinoma. In this chapter we describe the modification of a CPE-encoding vector by structure-directed mutagenesis to either preferentially target Cldn-1 and -5 or to expand targeting to Cldn1-9 for improved broadened cytotoxic targeting of claudin-overexpressing tumors such as but not limited to lung cancer via CPE gene transfer.


Assuntos
Carcinoma Pulmonar de Células não Pequenas , Neoplasias Pulmonares , Claudinas/genética , Claudinas/metabolismo , Clostridium perfringens/genética , Clostridium perfringens/metabolismo , Enterotoxinas/genética , Enterotoxinas/metabolismo , Terapia Genética , Humanos , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/terapia
9.
Poult Sci ; 101(6): 101848, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35544960

RESUMO

Several factors predisposing to necrotic enteritis (NE) have been identified, including diet and Eimeria spp. infestations. Coccidiosis vaccines are indicated to decrease the intestinal lesions caused by specific Eimeria species that are a known predisposing factor to NE and, consequently, these vaccines could be a holistic approach to the control of NE disease and an alternative solution to coccidiostats. Besides, feed additives have also gained special attention from the poultry industry as an alternative solution to antibiotics to prevent NE as well as other bacterial enteritis. Then, the combination of vaccination against coccidiosis and the supplementation of the diet with feed additives could be a composite approach to the control of NE problems triggered by Eimeria spp. infestation. The objective of this study was to test the efficacy of an attenuated coccidiosis vaccine (EVANT) in combination with different feed additives to prevent the loss of production performance and intestinal lesions in broilers challenged with NE. Healthy day-old broilers (n = 960) were randomly allocated to 6 groups (8 cages/group). Groups 1-2 were left unvaccinated. Groups 3-6 were vaccinated following the manufacturer's instructions. Chickens were grown using a diet favoring the intestinal proliferation of Clostridium perfringens. Moreover, the diets of groups 4-6 were supplemented with medium chain fatty acids (MCFA), butyric acid or phytogenic feed additives (PFA), respectively. A NE infection model was used to challenge groups 2-6; chickens were orally infected with Eimeria maxima (4,500 oocysts) and then C. perfringens (108 CFU) at 15 and 20 d, respectively. Birds were monitored and productive parameters recorded until 42 d; intestinal lesions were scored. Results showed that coccidiosis vaccination, with or without the addition of feed additives, decreased intestinal lesions associated with NE and improved the performance of the birds. Besides, the addition of MCFA to the diet decreased intestinal lesions associated to NE in vaccinated animals compared to all treatment groups. Moreover, the same additive improved the feed conversion rate. Therefore, vaccination with a live attenuated coccidiosis vaccine together with in-feed inclusion of MCFA might be a solution to reduce NE in broilers raised antimicrobial- and coccidiostat-free.


Assuntos
Infecções por Clostridium , Coccidiose , Eimeria , Enterite , Óleos Voláteis , Doenças das Aves Domésticas , Ração Animal/análise , Animais , Galinhas , Infecções por Clostridium/microbiologia , Infecções por Clostridium/prevenção & controle , Infecções por Clostridium/veterinária , Clostridium perfringens , Coccidiose/prevenção & controle , Coccidiose/veterinária , Dieta/veterinária , Enterite/microbiologia , Enterite/prevenção & controle , Enterite/veterinária , Doenças das Aves Domésticas/microbiologia , Vacinas Atenuadas
10.
Am J Case Rep ; 23: e936329, 2022 May 08.
Artigo em Inglês | MEDLINE | ID: mdl-35526110

RESUMO

BACKGROUND Clostridium perfringens (CP), one of several clostridial species gram-positive bacteria, is a major cause of animal necrosis enteritis and traumatic gangrene. In some reports, CP can cause acute emphysematous cholecystitis in patients with biliary tract infections. However, C. perfringens combined with other aerobic bacteria (eg, E. coli) in bloodstream co-infection is extremely rare and often fatal. Herein, we present a case of co-infection to underscore this unusual situation so that clinicians can adequately evaluate and treat patients in time. CASE REPORT A 74-year-old man presented to the Emergency Department half a day after the onset of acute abdominal pain accompanied by nausea, vomiting, and chills. The patient was admitted, following development of jaundice, chills, high fever, confusion, and shock. Computed tomography (CT) revealed that the patient had cholangiectasis with acute obstructive suppurative cholangitis (AOSC). We subsequently performed percutaneous transhepatic gallbladder drainage surgery combined with antibiotics, including ceftriaxone, levofloxacin, and metronidazole. C. perfringens and Escherichia coli infections were identified by in vitro blood culture. Fortunately, the patient responded favorably to treatment in our hospital and was cured within 1 week. CONCLUSIONS We report a rare case of C. perfringens and E. coli bloodstream co-infection in a patient with AOSC. We suggest that anaerobic and aerobic co-infection should be considered in future clinical diagnoses. Effective antibiotic treatment combined with surgical drainage is crucial if mixed infection occurs.


Assuntos
Bacteriemia , Colangite , Infecções por Clostridium , Coinfecção , Infecções por Escherichia coli , Sepse , Antibacterianos/uso terapêutico , Bacteriemia/complicações , Bacteriemia/diagnóstico , Bacteriemia/tratamento farmacológico , Calafrios , Colangite/complicações , Colangite/diagnóstico , Infecções por Clostridium/complicações , Infecções por Clostridium/diagnóstico , Infecções por Clostridium/microbiologia , Clostridium perfringens , Escherichia coli , Infecções por Escherichia coli/complicações , Infecções por Escherichia coli/diagnóstico , Humanos , Sepse/complicações , Supuração
11.
J Comp Pathol ; 194: 39-49, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35577458

RESUMO

Four toco toucans (Ramphastos toco), one channel-billed toucan (Ramphastos vitellinus) and one white-throated toucan (Ramphastos tucanus) died in two disease outbreaks in the same aviary in 2011 and 2016. Post-mortem examination revealed diffuse necrotic enteritis (NE) as the cause of death of five of these six birds. Clostridium perfringens was identified by culture and real-time multiplex PCR for C. perfringens α-, ß-, ε- and ι-toxin genes in ligated intestine of one toucan from each outbreak. At another aviary, two keel-billed toucans (Ramphastos sulfuratus) died peracutely from severe haemolytic crisis with haemoglobinaemic nephrosis and cholestasis and acute tubulointerstitial nephritis. Mild NE was present in these birds and C. perfringens was demonstrated in liver by bacterial culture and real-time multiplex PCR for C. perfringens α-, ß-, ε- and ι-toxin genes. To the authors' knowledge, this is the first description of outbreaks of NE associated with C. perfringens in captive toucans. Although haemolytic crisis has been reported in humans with C. perfringens type A septicaemia and hepatic abscesses, this presentation appears not to have been described in C. perfringens infections in toucans or other avian species. The factors causing C. perfringens proliferation and disease in the toucans were not identified. PCR for C. perfringens NetB toxin and enterotoxin genes performed retrospectively on one of the C. perfringens isolates from the second outbreak and on paraffin-embedded tissues from one dead toucan from the first outbreak was negative. With the current C. perfringens toxin typing scheme, C. perfringens type A was identified in the first two outbreaks.


Assuntos
Infecções por Clostridium , Enterite , Doenças das Aves Domésticas , Animais , Galinhas , Infecções por Clostridium/epidemiologia , Infecções por Clostridium/microbiologia , Infecções por Clostridium/veterinária , Clostridium perfringens/genética , Surtos de Doenças/veterinária , Enterite/epidemiologia , Enterite/veterinária , Doenças das Aves Domésticas/epidemiologia , Estudos Retrospectivos
12.
Acta Trop ; 232: 106537, 2022 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-35623400

RESUMO

Although some studies on the effects of para-probiotics on the immune system and intestinal health have been conducted independently of research on antibiotics ass growth promoters. This study investigated the effects of heat-killed Lactobacillus plantarumL-137 (L-137) and antibiotics as preventive and/or therapeutic substances for broilers against subclinical necrotic enteritis caused by Clostridium perfringens (CP). In total, 300 1-day-old broilers (46.13 ± 1.38 g) were randomly stocked at 10 birds pen-1 in five replicates and divided into six groups, namely T1 and T2, positive and negative control of CP challenge; T3 and T4, prevention with basal diet plus 10 and 50 mg/kg L-137; T5 and T6, prevention and treatment with basal diet plus 50 mg/kg of L-137 and bacitracin at 50 ppm, respectively. Broilers administered L-137 in T4, T5 and bacitracin in T6 showed an improved (p < 0.05) villus height/crypt depth ratio than control groups, suggesting that it might significantly boost growth performance. In contrast to bacitracin, a high dosage of L-137 significantly increased (p < 0.05) the spleen index value and the cytokine levels, as well as the expression of intestinal ß-defensin genes on day 28. During the 42-day production period, broilers in T4 and T5 showed a significantly enhanced (p < 0.05) expression of cytokines, AvBD-1 and AvBD-7 on day 42 compared to the control and bacitracin groups. In particular, broilers given the L-137 diets demonstrated no cumulative mortality following CP exposure, compared to a 2% mortality in T6. Our findings provide insight into eco-friendly alternatives to antibiotics for maximizing growth performance, feed efficiency and long-term disease protection in chickens; however, this has to be proven in larger-scale commercial experiments.


Assuntos
Lactobacillus plantarum , Doenças das Aves Domésticas , Ração Animal/análise , Animais , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Bacitracina/farmacologia , Bacitracina/uso terapêutico , Galinhas , Clostridium perfringens , Dieta/veterinária , Suplementos Nutricionais/análise , Temperatura Alta , Doenças das Aves Domésticas/prevenção & controle
13.
Microbiologyopen ; 11(2): e1270, 2022 04.
Artigo em Inglês | MEDLINE | ID: mdl-35478283

RESUMO

Necrotic enteritis (NE), caused by Clostridium perfringens, is an intestinal disease with devastating economic losses to the poultry industry. NE is a complex disease and predisposing factors that compromise gut integrity are required to facilitate C. perfringens proliferation and toxin production. NE is also characterized by drastic shifts in gut microbiota; C. perfringens is negatively correlated with Lactobacilli. Vaccines are only partially effective against NE and antibiotics suffer from the concern of resistance development. These strategies address only some aspects of NE pathogenesis. Thus, there is an urgent need for alternative strategies that address multiple aspects of NE biology. Here, we developed Limosilactobacillus (Lactobacillus) reuteri vectors for in situ delivery of nanobodies against NetB and α toxin, two key toxins associated with NE pathophysiology. We generated nanobodies and showed that these nanobodies neutralize NetB and α toxin. We selected L. reuteri vector strains with intrinsic benefits and demonstrated that these strains inhibit C. perfringens and secrete over 130 metabolites, some of which play a key role in maintaining gut health. Recombinant L. reuteri strains efficiently secreted nanobodies and these nanobodies neutralized NetB. The recombinant strains were genetically and phenotypically stable over 480 generations and showed persistent colonization in chickens. A two-dose in ovo and drinking water administration of recombinant L. reuteri strains protected chickens from NE-associated mortality. These results provide proof-of-concept data for using L. reuteri as a live vector for delivery of nanobodies with broad applicability to other targets and highlight the potential synergistic effects of vector strains and nanobodies for addressing complex diseases such as NE.


Assuntos
Toxinas Bacterianas , Infecções por Clostridium , Enterite , Doenças das Aves Domésticas , Anticorpos de Domínio Único , Animais , Toxinas Bacterianas/genética , Toxinas Bacterianas/metabolismo , Galinhas , Infecções por Clostridium/patologia , Infecções por Clostridium/prevenção & controle , Infecções por Clostridium/veterinária , Clostridium perfringens/genética , Clostridium perfringens/metabolismo , Enterite/prevenção & controle , Enterite/veterinária , Enterotoxinas/genética , Enterotoxinas/metabolismo , Lactobacillus/metabolismo , Doenças das Aves Domésticas/prevenção & controle , Anticorpos de Domínio Único/genética , Anticorpos de Domínio Único/metabolismo
14.
Toxins (Basel) ; 14(4)2022 04 16.
Artigo em Inglês | MEDLINE | ID: mdl-35448898

RESUMO

Epsilon toxin (Etx) from Clostridium perfringens is the third most potent toxin after the botulinum and tetanus toxins. Etx is the main agent of enterotoxemia in ruminants and is produced by Clostridium perfringens toxinotypes B and D, causing great economic losses. Etx selectively binds to target cells, oligomerizes and inserts into the plasma membrane, and forms pores. A series of mutants have been previously generated to understand the cellular and molecular mechanisms of the toxin and to obtain valid molecular tools for effective vaccination protocols. Here, two new non-toxic Etx mutants were generated by selective deletions in the binding (Etx-ΔS188-F196) or insertion (Etx-ΔV108-F135) domains of the toxin. As expected, our results showed that Etx-ΔS188-F196 did not exhibit the usual Etx binding pattern but surprisingly recognized specifically an O-glycoprotein present in the proximal tubules of the kidneys in a wide range of animals, including ruminants. Although diminished, Etx-ΔV108-F135 maintained the capacity for binding and even oligomerization, indicating that the mutation particularly affected the pore-forming ability of the toxin.


Assuntos
Clostridium perfringens , Enterotoxemia , Animais , Sítios de Ligação , Membrana Celular/metabolismo , Clostridium perfringens/genética , Clostridium perfringens/metabolismo , Enterotoxemia/genética , Ligação Proteica
15.
Viruses ; 14(4)2022 03 25.
Artigo em Inglês | MEDLINE | ID: mdl-35458406

RESUMO

Clostridium perfringens is an important pathogen for both humans and animals, causing human foodborne disease and necrotic enteritis in poultry. In the present study, a C. perfringens-specific phage, vB_CpeS_BG3P (designated as BG3P hereafter), was isolated from chicken farm sewage. Both electron microscopy and phylogenetic analysis suggested that phage BG3P is a novel phage belonging to Siphoviridae family. Phage BG3P exhibited a broad host range against different C. perfringens isolates (90.63% of strains were infected). Sequencing of the complete genome revealed a linear double-stranded DNA (43,528 bp) with 28.65% GC content. After sequence analysis, 73 open reading frames (orfs) were predicted, of which only 13 were annotated with known functions. No tRNA and virulence encoding genes were detected. It should be noted that the protein of orf 15 has 97.92% homology to C. perfringens-specific chloramphenicol resistance protein, which has not been reported for any C. perfringens phage. Phylogenetic analysis of the ssDNA binding protein demonstrated that this phage is closely related to C. perfringens phages phiSM101 and phi3626. In considering future use as an antimicrobial agent, some biological characteristics were observed, such as a good pH (3-11) stability and moderate temperature tolerance (<60 °C). Moreover, bacteriophage BG3P showed a good antimicrobial effect against C. perfringens liquid cultures. Thus, phage treatment with MOI ≥ 100 completely inhibited bacterial growth compared to untreated cultures. Although phage BG3P shows good lytic efficiency and broad host range in vitro, future development and application may need to consider removal of the chloramphenicol-like resistance gene or exploring its lysin for future antibacterial applications.


Assuntos
Bacteriófagos , Siphoviridae , Animais , Clostridium perfringens/genética , Genoma Viral , Especificidade de Hospedeiro , Filogenia , Siphoviridae/genética
16.
Avian Dis ; 66(1): 1-4, 2022 03.
Artigo em Inglês | MEDLINE | ID: mdl-35366673

RESUMO

Necrotic enteritis (NE) is a serious disease of chickens and turkeys that causes significant economic losses to the poultry industry. On the basis of studies in chickens, Clostridium perfringens type G is considered by many to be the cause of NE in poultry. However, studies on isolates from Finnish and Italian turkeys with NE revealed that the vast majority were C. perfringens type A, and very few were C. perfringens type G. We therefore examined 74 C. perfringens isolates from U.S. turkeys with NE; 98% were type A and only 1% was type G. This result confirms that different C. perfringens types are involved in NE in turkeys when compared with chickens. We also examined the turkey isolates for other toxin genes associated with enteritis in various animal species, namely tpeL, cpb2, cpe, netE, netF, and netG. The tpeL gene, which has been associated with enhanced virulence of C. perfringens in chickens, was only found in 1% of turkey NE isolates. The cpe gene, which encodes C. perfringens enterotoxin (a major cause of food poisoning and non-foodborne C. perfringens-mediated diarrhea in humans) was also found in only 1% of our turkey NE isolates. Although cpb2, which encodes for the beta2 toxin, was found in 73% of our NE isolates, it has also been found in similar percentages of isolates from turkeys with normal intestine. The netE, netF, and netG genes were not detected among our C. perfringens isolates from turkeys.


Reporte de caso- Tipificación de toxinas de cepas de Clostridium perfringens recuperadas de pavos con enteritis necrótica en los Estados Unidos. La enteritis necrótica es una enfermedad severa de los pollos y pavos que provoca importantes pérdidas económicas a la industria avícola. Sobre la base de estudios en pollos, muchos consideran que Clostridium perfringens tipo G es la causa de la enteritis necrótica en las aves comerciales. Sin embargo, los estudios sobre los aislamientos de pavos con enteritis necrótica de Finlandia e Italia revelaron que la gran mayoría de los aislamientos eran C. perfringens tipo A y muy pocos eran C. perfringens tipo G. Por lo tanto, se examinaron 74 aislamientos de C. perfringens de pavos en los Estados Unidos con enteritis necrótica. El 98% eran del tipo A y solo el 1% fueron del tipo G. Este resultado confirma que diferentes tipos de C. perfringens están involucrados en la enteritis necrótica en pavos en comparación con los pollos. También se examinaron los aislamientos de pavo en busca de otros genes de toxinas asociados con la enteritis en varias especies animales, especialmente, tpeL, cpb2, cpe, netE, netF y netG. El gene tpeL, que se ha asociado con una mayor virulencia de C. perfringens en pollos, solo se encontró en el 1 % de los aislamientos de enteritis necrótica de pavo. El gene cpe, que codifica la enterotoxina de C. perfringens (una de las principales causas de intoxicación alimentaria y diarrea no transmitida por los alimentos causada por C. perfringens en humanos) también se encontró en solo el 1 % de los aislamientos de enteritis necrótica de pavo. Aunque cpb2, que codifica para la toxina beta2, se encontró en el 73 % de los aislamientos de enteritis necrótica, también se ha encontrado en porcentajes similares de aislamientos de pavos con intestino normal. Los genes netE, netF y netG no se detectaron entre los aislamientos de C. perfringens de pavos.


Assuntos
Infecções por Clostridium , Enterite , Doenças das Aves Domésticas , Animais , Galinhas , Infecções por Clostridium/veterinária , Clostridium perfringens , Enterite/veterinária , Enterotoxinas/genética , Aves Domésticas , Perus
17.
Poult Sci ; 101(4): 101720, 2022 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-35231770

RESUMO

The purpose of this experiment is to explore the effects of dietary supplementation of benzoic acid, Enterococcus faecium, and essential oil complex (BEC) on coccidia and Clostridium perfringens challenge in laying hens. A total of 80 Lohmann gray laying hens (35 wk old) were allocated to 4 treatments in a 2 × 2 factorial arrangement with the main effects of Clostridium perfringens type A (CP) and coccidia challenge (with or without challenge) and 2 BEC levels (0 and 1,000 mg/kg). The total experimental period was 6 wk. The results showed that: the challenge group significantly decreased the laying rate and average daily feed intake (ADFI) of laying hens (PChallenge < 0.01). The BEC + challenge group significantly increased the laying rate and decreased the feed conversion ratio (FCR) of laying hens (PBEC < 0.05). The challenge significantly decreased the thickness, strength, and relative weight of eggshell (PChallenge < 0.05). The BCE + challenge group significantly increased the relative weight and strength of the eggshell (PBEC < 0.05). The challenge significantly increased the crypt depth of the duodenum, jejunum and ileum, and decreased the villus-to-crypt ratio (V/C) (PChallenge < 0.01). The BEC + challenge group decreased the crypt depth of the duodenum and jejunum, and increased the V/C of the duodenum (PBEC < 0.01). The pathological scores of duodenum and jejunum of the challenge group were significantly higher than other groups (PChallenge < 0.01), while the BEC + challenge group had lower pathological scores of jejunum (PBEC < 0.01). The challenge significantly decreased the mRNA expression of Occludin, Mucin-2, Zonula occluden-1 (ZO-1) (Pchallenge < 0.05); whereas the BEC group significantly increased the expression of Occludin, Mucin-2, and Claudin-1 mRNA (PBEC < 0.05). The challenge significantly increased the level of interleukin 1ß (IL-1ß) in the jejunum (PChallenge < 0.05). Taken together, adding BEC to the diet can improved production performance and egg quality of layers, by protecting intestinal health against Clostridium perfringens type A (CP) and coccidia challenge.


Assuntos
Coccídios , Enterococcus faecium , Óleos Voláteis , Ração Animal/análise , Fenômenos Fisiológicos da Nutrição Animal , Animais , Ácido Benzoico , Galinhas , Clostridium perfringens , Dieta/veterinária , Suplementos Nutricionais/análise , Feminino , Mucina-2 , Ocludina , Óleos Voláteis/farmacologia , Óvulo , RNA Mensageiro
18.
Avian Dis ; 66(1): 1-8, 2022 03.
Artigo em Inglês | MEDLINE | ID: mdl-35308011

RESUMO

Necrotic enteritis (NE) is an important intestinal disease of commercial poultry associated with poor performance, high mortality, and significant economic loss. In this case report, a novel presentation of NE is described in young broilers. Initially, affected farms presented with a moderate increase in mortality at or before 19 days of age. Farms experiencing the disease syndrome belonged to two complexes of the same company. However, all farms sourced chicks from the same hatchery. Farm postmortem examinations revealed moderate to severe enteritis of the upper small intestine characterized by multifocal, irregular, plaque-like, mucosal ulcerations. Additionally, thinning of the intestinal wall with consequential distension and ballooning and a necrotic, pseudomembranous layer covering the mucosa were observed in some birds. Clinically affected birds were submitted to the Poultry Research and Diagnostic Laboratory at Mississippi State University for further evaluation. Birds were between 5 and 11 days of age and presented with similar gross lesions. Anaerobic culture was performed, and Clostridium perfringens was isolated from affected intestinal sections. Environmental sampling at the hatchery was also performed to evaluate the presence and load of clostridial organisms. Clostridium perfringens was isolated from samples collected in the egg room, hatchers/hatch halls, separator room, processing room, and transport trucks. Furthermore, VITEK® mass spectrometry matrix-assisted laser desorption ionization time-of-flight results indicated a 99.9% genetic relatedness between hatchery and live bird isolates, demonstrating an epidemiologic link between clinically affected birds and the hatchery as a point source. This novel presentation of C. perfringens in young broilers warrants attention because proper development of the gastrointestinal tract in the first weeks of life is critical for competitive production in the modern broiler.


Reporte de caso- Nueva presentación de Clostridium perfringens en pollos de engorde jóvenes. La enteritis necrótica es una enfermedad intestinal importante de las aves comerciales asociada con un bajo rendimiento, alta mortalidad y pérdidas económicas significativas. En este reporte de caso, se describe una presentación nueva de enteritis necrótica en pollos de engorde jóvenes. Inicialmente, las granjas afectadas presentaron un aumento moderado en la mortalidad a los 19 días de edad o antes. Las granjas que presentaban el síndrome de la enfermedad pertenecían a dos complejos de la misma empresa. Sin embargo, todas las granjas obtuvieron pollitos de la misma incubadora. Los exámenes post mortem de la granja revelaron una enteritis de moderada a severa en la parte superior del intestino delgado caracterizada por ulceraciones de la mucosa multifocales, irregulares y similares a placas. Además, en algunas aves se observó adelgazamiento de la pared intestinal con la consiguiente distensión e hinchazón y una capa pseudomembranosa necrótica que cubría la mucosa. Las aves clínicamente afectadas se enviaron al Laboratorio de Investigación y Diagnóstico Avícola de la Universidad Estatal de Mississippi para una evaluación adicional. Las aves tenían entre cinco y once días de edad y presentaban lesiones macroscópicas similares. Se realizó cultivo de anaerobios y se aisló Clostridium perfringens de las secciones intestinales afectadas. También se realizaron muestreos ambientales en la incubadora para evaluar la presencia y carga de organismos clostridiales. Se aisló Clostridium perfringens de muestras recolectadas en la sala de huevos, incubadoras/salones de incubación, sala de separación, sala de procesamiento y camiones de transporte. Además, los resultados de la espectrometría de masas MALDI-TOF de VITEK® indicaron una relación genética del 99.9 % entre los aislados de la incubadora y de aves vivas, lo que demuestra un vínculo epidemiológico entre las aves clínicamente afectadas y la incubadora como fuente de infección común. Esta nueva presentación de C. perfringens en pollos de engorde jóvenes merece atención porque el desarrollo adecuado del tracto gastrointestinal en las primeras semanas de vida es fundamental para la producción competitiva en el pollo de engorde moderno.


Assuntos
Infecções por Clostridium , Enterite , Doenças das Aves Domésticas , Animais , Galinhas , Infecções por Clostridium/diagnóstico , Infecções por Clostridium/veterinária , Clostridium perfringens , Enterite/patologia , Enterite/veterinária , Humanos , Necrose/veterinária , Doenças das Aves Domésticas/diagnóstico , Doenças das Aves Domésticas/patologia
19.
Poult Sci ; 101(4): 101751, 2022 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-35240353

RESUMO

Akkermansia muciniphila (AM) is a mucin-degrading anaerobe, exerting beneficial effects on gut integrity improvement, inflammatory alleviation, and metabolic regulations in humans. Excess amounts of mucin and mucogenesis in the gut facilitate the development of necrotic enteritis (NE) in chickens. The study aimed to evaluate the effects of oral inoculation of AM on NE prevention and gut modulation in a NE-reproduced model coinfecting with Clostridium perfringens (CP) and Eimeria parasites. A total of 105 commercial 1-day-old broilers were randomly allocated into 5 groups, respectively challenged with Eimeria (Eimeria group), Eimeria and CP (Eimeria+CP group), Eimeria and CP with AM (Eimeria+CP+AM group), Eimeria and AM (Eimeria+AM group), and a placebo (Noninfected group). The treatment of AM exhibited a low degree of amelioration on NE severity. The application neither protected broilers from NE by decreasing NE-positive numbers nor reached a significant reduction in lesion scores in the small intestines. The development of NE reduced species diversity in jejunal microbiota; the pretreatments of AM exacerbated the consequence by losing species richness and promoted the similarity of the jejunal microbial community presented in the Eimeria+CP group. The participation of AM enhanced the increments of genera Clostridium sensu stricto 1 and Escherichia_Shigella and decreased the number of Lactobacillus. The significant variations of genera Clostridium sensu stricto 1 and Lactobacillus in jejunal microbiota were associated with NE development and promotion. In conclusion, oral inoculation of AM promoted the development of NE and modulated the jejunal microbiota favorable for CP overgrowth in broilers. The application of AM as a probiotic in broilers should be cautious on account of the effects to predispose NE.


Assuntos
Infecções por Clostridium , Coccidiose , Eimeria , Enterite , Microbioma Gastrointestinal , Doenças das Aves Domésticas , Ração Animal/análise , Animais , Galinhas , Infecções por Clostridium/patologia , Infecções por Clostridium/prevenção & controle , Infecções por Clostridium/veterinária , Clostridium perfringens , Coccidiose/parasitologia , Coccidiose/prevenção & controle , Coccidiose/veterinária , Enterite/patologia , Enterite/prevenção & controle , Enterite/veterinária , Incidência , Mucinas , Doenças das Aves Domésticas/parasitologia , Doenças das Aves Domésticas/prevenção & controle
20.
Poult Sci ; 101(4): 101753, 2022 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-35240358

RESUMO

Necrotic enteritis (NE) is a devastating disease that has seen a resurgence of cases following the removal of antibiotics from feed resulting in financial loss and significant animal health concerns across the poultry industry. The objective was to evaluate the efficacy of a microencapsulated blend of organic (25% citric and 16.7% sorbic) acids and botanicals (1.7% thymol and 1% vanillin [AviPlusP]) to reduce clinical NE and determine the signaling pathways associated with any changes. Day-of-hatch by-product broiler breeder chicks were randomly assigned to a control (0) or supplemented (500 g/MT) diet (n = 23-26) and evaluated in a NE challenge model (n = 3). Birds were administered 2X cocci vaccine on d 14 and challenged with a cocktail of Clostridium perfringens strains (107) on d 17 to 19. On d 20 to 21 birds were weighed, euthanized, and scored for NE lesions. Jejunal tissue was collected for kinome analysis using an immuno-metabolism peptide array (n = 5; 15/treatment) to compare tissue from supplement-fed birds to controls. Mortality and weight were analyzed using Student's t test and lesion scores analyzed using F-test two-sample for variances (P < 0.05). The kinome data was analyzed using PIIKA2 peptide array analysis software and fold-change between control and treated groups determined. Mortality in the supplemented group was 47.4% and 70.7% in controls (P = 0.004). Lesions scores were lower (P = 0.006) in supplemented birds (2.47) compared to controls (3.3). Supplement-fed birds tended (P = 0.19) to be heavier (848.6 g) than controls (796.2 g). Kinome analysis showed T cell receptor, TNF and NF-kB signaling pathways contributed to the improvements seen in the supplement-fed birds. The following peptides were significant (P < 0.05) in all 3 pathways: CHUK, MAP3K14, MAP3K7, and NFKB1 indicating their importance. Additionally, there were changes to IL6, IL10, and IFN- γ mRNA expression in tissue between control- and supplement-fed chickens. In conclusion, the addition of a microencapsulated blend of organic acids and botanicals to a broiler diet reduced the clinical signs of NE that was mediated by specific immune-related pathways.


Assuntos
Infecções por Clostridium , Enterite , Doenças das Aves Domésticas , Ácidos , Ração Animal/análise , Animais , Galinhas , Infecções por Clostridium/prevenção & controle , Infecções por Clostridium/veterinária , Clostridium perfringens , Dieta/veterinária , Enterite/tratamento farmacológico , Enterite/prevenção & controle , Enterite/veterinária , Humanos , Necrose/prevenção & controle , Necrose/veterinária , Compostos Orgânicos , Doenças das Aves Domésticas/prevenção & controle , Transdução de Sinais
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