A Comparative Study of the Effect of Non-Antibiotic Feed Additives on Experimental Colonization of Salmonella Enterica Serovar Enteridis and Intestinal Pathomorphology in Broiler Chickens

The objective of this study was to evaluate the effect of eubiotics on the intestinal morphology of broilers. For this purpose, 125 birds were divided into six groups with two replicates each (10 birds in each replicate). Group A was given a Basal diet. All groups except group A were challenged with Salmonella enterica serovar Enteritidis. Group B was provided the basal diet, group C was fed a Probiotic-added diet; group D was fed a Prebiotics-based diet; group E was given essential oils plus the basal diet; and group F was provided with organic acids plus the basal diet. Two separate experiments were carried out for Salmonella recovery, checking the cecal tonsils and conducting an intestinal pathomorphic evaluation. Villus length, villus width, villus surface area, and crypt depth were measured by micrometry. There was an overall improvement (p<0.05) in intestinal morphometric parameters for all the treatment groups except for the negative control group, which showed the lowest villus height and villus depth values. Maximum villus height (p<0.05) of the duodenum was achieved by group E, which was fed a diet containing essential oils, whereas a maximum villus surface area index (p<0.05) was recorded for the birds of Group D, which were fed a diet containing prebiotics. Maximum villus height (p<0.05) and surface area index in ileum mucosa was recorded (p<0.05) in the birds of group D (treated with prebiotics). It is concluded that there is an overall increase in the gut histology of broilers fed non-antibiotic based feed.(AU)

Effect of Bacillus Subtilis on Immune Function of Hd11 Chicken Macrophages

Poultry is frequently contaminated by Salmonella, a pathogen leading to human health concern worldwide. This study aimed to evaluate the effect of Bacillus subtilis (BS)strain 048 (BS048) on the activation, phagocytosis, sterilization, cytokine secretion, and nitrogen oxide synthesis of HD11 chicken macrophages subjected to Salmonella enteritidis challenge, using lipopolysaccharide treatment as a negative control. The results showed: (1) BS048 had no significant effect on extracellular lactate dehydrogenase activity (p>0.05), while lipopolysaccharide treatment significantly increased extracellular lactate dehydrogenase activity (p >0.05), while lipopolysaccharide treatment significantly increased extracellular lactate dehydrogenase activity (p 0.05);(2)BS048 significantly upregulated the expression levels of pro-inflammatory cytokines (interleukin (IL)- 1â and IL-6), anti-inflammatory cytokines(IL-10 and transforming growth factor-â1), and anti-viral cytokine, interferon-â (p<0.01); ; (3) BS048 significantly upregulated the mRNA expression level of the inducible nitric oxide synthase and its activity as well as extracellular nitrogen oxide level (p <0.01). In conclusion, BS048 could improve antiinflammatory and immune functions of HD11 chicken macrophages, without cytotoxic effects on these cells.(AU)

Evaluation of Analytical Sensitivity of Sdf I based PCR and Sandwich ELISA for Salmonella Enteritidis detection and On-Farm prevalence in Punjab, Pakistan

Salmonella Enteritidis (SE) is a dominant serotype among non-typhoidal Salmonella which renders poultry products unsafe for human consumption. Due to frequent reporting of egg associated outbreaks, broiler breeder flocks are understudied although farm environment present supporting conditions for the growth of SE. In this study, two rapid detection techniques for SE were compared in terms of analytical sensitivity and the extent of SE contamination in broiler breeder farm environment was determined. Analytical sensitivity as limit of detection (LOD) was evaluated quantitatively for serotype specific PCR based on amplification of Sdf I gene and a commercially available sandwich ELISA for antigen detection. In triplicate experiments, tenfold serial dilutions of SE were prepared and tested with each technique. Using pure cultures, analytical sensitivity of PCR and ELISA were found to be 18.6 CFU/ml and 2.77×105 CFU/ml respectively. PCR (LOD, log 1.2) was found to be more sensitive and rapid than ELISA (LOD, log 5.4). Environmental swab samples (n = 260) were collected from 22 hen houses representing 8 broiler breeder farms located in and around Lahore and Sheikhupura districts of Punjab province. From each hen house swab samples were collected from litter, nests, feeders, drinkers, fans, pads, ceiling, walls and walkways. Following selective enrichment, pooled swab samples were subjected to PCR. Results showed that 36.3 % (8/22) hen houses were detected positive for SE. These findings suggest improvement in farm biosecurity measures and advocate implementation of integrated Salmonellosis control programs in broiler breeder houses to minimize carcass contamination.(AU)

Bacillus coagulans as an alternative to antibiotics in the performance and control of Salmonella Enteritidis in broilers / Bacillus coagulans como alternativa aos antibióticos sobre o desempenho e controle da Salmonella Enteritidis em frangos de corte

The use of antimicrobial growth promoters in broiler feed has been beneficial for improving performance and preventing diseases. However, the indiscriminate use of these products in the feed can result in the development of resistant bacteria, the accumulation of residues in the products, and an imbalance in the microflora of birds. Therefore, it is necessary to evaluate alternatives, such as beneficial microorganisms that improve microbial growth without affecting animal health and product quality. This research aimed to evaluate the supplementation with the probiotic Bacillus coagulans on the performance, carcass characteristics, and health of broilers from seven to 42 days. In total, 720 broilers were used, distributed in a randomized block design with six treatments and eight replicates. The evaluated treatments were as follows: Control ration (RC); PROB1 (Probiotic 400 g/t); PROB2 (Probiotic 400 g/t until 21 days and 200 g/t from 22 to 42 days); RC + antibiotic; RC + Salmonella inoculation; PROB1 + Salmonella inoculation. The treatments did neither influence feed intake, carcass yield, and cuts nor the incidence of injuries to the chest, hock, and footpad. Weight gain and feed conversion were better in birds that received antibiotic or probiotic diets. There was an incidence of Salmonella in the challenges excreta at42 days only in the treatment with challenge without adding probiotics. We conclude that the probiotic Bacillus coagulans can be used as an alternative to antibiotics in the diet of broilers as it facilitates similar performance and is efficient in the control of Salmonella Enteritidis.

A utilização de promotores de crescimento antimicrobianos na alimentação de frangos de corte tem sido benéfica para melhoria do desempenho e para prevenção de doenças. Porém, o uso indiscriminado destes produtos nas rações pode resultar em desenvolvimento de bactérias resistentes, acúmulo de resíduos nos produtos e desequilíbrio da microflora das aves. Portanto, torna-se necessário avaliar alternativas como microrganismos benéficos que melhorem o crescimento microbiano, sem afetar a saúde do animal e a qualidade dos seus produtos. Objetivou-se com esta pesquisa avaliar a suplementação do probiótico Bacillus coagulans sobre o desempenho, características de carcaça e saúde de frangos de corte de 7 a 42 dias. Utilizou-se 720 frangos de corte distribuídos em delineamento em blocos casualizados, com seis tratamentos e oito repetições. Os tratamentos avaliados foram: Controle; PROB1 (Probiótico 400 g/t); PROB2 (Probiótico 400 g/ton até os 21 dias e 200 g/t dos 22 aos 42 dias); Controle + antibiótico; Controle + inoculação de Salmonella; PROB1 + inoculação de Salmonella. Os tratamentos não influenciaram o consumo de ração, rendimento de carcaça e cortes e a incidência de lesões no peito, jarrete e coxim plantar. O ganho de peso e a conversão alimentar foram melhores nas aves que receberam rações com antibiótico ou probiótico. Houve incidência de Salmonella nas excretas aos 42 dias somente no tratamento com desafio sem adição de probiótico. Conclui-se que o probiótico Bacillus coagulans pode ser usado como alternativa ao antibiótico na ração de frangos de corte, pois proporciona desempenho semelhante e é eficiente no controle da Salmonella Enteritidis

Antimicrobial and insecticidal effects of essential oil and plant extracts of Myrcia oblongata DC in pathogenic bacteria and Alphitobius diaperinus

Abstract The secondary metabolism products of plants have influenced great economic interest, given their chemical diversity and biological activities. Because of this, this study evaluates the phytochemical composition, antimicrobial activity, insecticidal, and antioxidant activity of plant extracts and oil of Myrcia oblongata. Saponins, steroids, triterpenoids, tannins, and flavonoids were detected. The extracts showed antimicrobial capacity on the tested microorganisms, except for the methanolic extract, which showed no activity for P. mirabilis and S. enteritidis. Regarding the analysis of antioxidant compounds, the hexanic, ethyl acetate and acetone extracts showed higher antioxidant activities and also higher insecticidal performance on Alphitobius diaperinus larvae, resulting in 80% adult mortality. The results reported here show that there may be a relationship between antioxidant potential and the insecticidal effect of Myrcia oblongata DC. The components present in both the extract and the oil can be used as natural alternative to synthetic compounds in the biological control of parasites and pathogenic microorganisms.

Resumo Os produtos do metabolismo secundário das plantas têm despertado grande interesse econômico, dada sua diversidade química e atividades biológicas. Neste sentido, o estudo objetivou avaliar a composição fitoquímica, atividade antimicrobiana, inseticida e antioxidante dos extratos vegetais e óleo de Myrcia oblongata. Foram detectados a presença de saponinas, esteróides, triterpenóides, taninos e flavonóides. Os extratos apresentaram capacidade antimicrobiana sobre os microrganismos testados, exceto o extrato metanólico que não demonstrou atividade para P. mirabilis e S. Enteritidis. Quanto a análise de compostos antioxidantes observou-se que os extratos hexânico, acetato de etila e acetona apresentaram maiores atividades antioxidantes e também maior performance inseticida sobre a larva Alphitobius diaperinus e exibindo mortalidade de 80% na fase adulta. Os resultados aqui reportados mostram que pode haver uma relação entre potencial antioxidante e efeito inseticida do óleo de Myrcia oblongata; os componentes presentes tanto no extrato como o oléo podem ser utilizados como alternativa natural aos compostos sintéticos no controle biológico de parasitas e microrganismos patogênicos.

Antimicrobial and insecticidal effects of essential oil and plant extracts of Myrcia oblongata DC in pathogenic bacteria and Alphitobius diaperinus / Aplicação de óleo e extratos de Myrcia oblongata DC em bactérias patogênicas e controle biológico de Alphitobius diaperinus

The secondary metabolism products of plants have influenced great economic interest, given their chemical diversity and biological activities. Because of this, this study evaluates the phytochemical composition, antimicrobial activity, insecticidal, and antioxidant activity of plant extracts and oil of Myrcia oblongata. Saponins, steroids, triterpenoids, tannins, and flavonoids were detected. The extracts showed antimicrobial capacity on the tested microorganisms, except for the methanolic extract, which showed no activity for P. mirabilis and S. enteritidis. Regarding the analysis of antioxidant compounds, the hexanic, ethyl acetate and acetone extracts showed higher antioxidant activities and also higher insecticidal performance on Alphitobius diaperinus larvae, resulting in 80% adult mortality. The results reported here show that there may be a relationship between antioxidant potential and the insecticidal effect of Myrcia oblongata DC. The components present in both the extract and the oil can be used as natural alternative to synthetic compounds in the biological control of parasites and pathogenic microorganisms.

Os produtos do metabolismo secundário das plantas têm despertado grande interesse econômico, dada sua diversidade química e atividades biológicas. Neste sentido, o estudo objetivou avaliar a composição fitoquímica, atividade antimicrobiana, inseticida e antioxidante dos extratos vegetais e óleo de Myrcia oblongata. Foram detectados a presença de saponinas, esteróides, triterpenóides, taninos e flavonóides. Os extratos apresentaram capacidade antimicrobiana sobre os microrganismos testados, exceto o extrato metanólico que não demonstrou atividade para P. mirabilis e S. Enteritidis. Quanto a análise de compostos antioxidantes observou-se que os extratos hexânico, acetato de etila e acetona apresentaram maiores atividades antioxidantes e também maior performance inseticida sobre a larva Alphitobius diaperinus e exibindo mortalidade de 80% na fase adulta. Os resultados aqui reportados mostram que pode haver uma relação entre potencial antioxidante e efeito inseticida do óleo de Myrcia oblongata; os componentes presentes tanto no extrato como o oléo podem ser utilizados como alternativa natural aos compostos sintéticos no controle biológico de parasitas e microrganismos patogênicos.

Antimicrobial and insecticidal effects of essential oil and plant extracts of Myrcia oblongata DC in pathogenic bacteria and Alphitobius diaperinus / Aplicação de óleo e extratos de Myrcia oblongata DC em bactérias patogênicas e controle biológico de Alphitobius diaperinus

The secondary metabolism products of plants have influenced great economic interest, given their chemical diversity and biological activities. Because of this, this study evaluates the phytochemical composition, antimicrobial activity, insecticidal, and antioxidant activity of plant extracts and oil of Myrcia oblongata. Saponins, steroids, triterpenoids, tannins, and flavonoids were detected. The extracts showed antimicrobial capacity on the tested microorganisms, except for the methanolic extract, which showed no activity for P. mirabilis and S. enteritidis. Regarding the analysis of antioxidant compounds, the hexanic, ethyl acetate and acetone extracts showed higher antioxidant activities and also higher insecticidal performance on Alphitobius diaperinus larvae, resulting in 80% adult mortality. The results reported here show that there may be a relationship between antioxidant potential and the insecticidal effect of Myrcia oblongata DC. The components present in both the extract and the oil can be used as natural alternative to synthetic compounds in the biological control of parasites and pathogenic microorganisms.(AU)

Os produtos do metabolismo secundário das plantas têm despertado grande interesse econômico, dada sua diversidade química e atividades biológicas. Neste sentido, o estudo objetivou avaliar a composição fitoquímica, atividade antimicrobiana, inseticida e antioxidante dos extratos vegetais e óleo de Myrcia oblongata. Foram detectados a presença de saponinas, esteróides, triterpenóides, taninos e flavonóides. Os extratos apresentaram capacidade antimicrobiana sobre os microrganismos testados, exceto o extrato metanólico que não demonstrou atividade para P. mirabilis e S. Enteritidis. Quanto a análise de compostos antioxidantes observou-se que os extratos hexânico, acetato de etila e acetona apresentaram maiores atividades antioxidantes e também maior performance inseticida sobre a larva Alphitobius diaperinus e exibindo mortalidade de 80% na fase adulta. Os resultados aqui reportados mostram que pode haver uma relação entre potencial antioxidante e efeito inseticida do óleo de Myrcia oblongata; os componentes presentes tanto no extrato como o oléo podem ser utilizados como alternativa natural aos compostos sintéticos no controle biológico de parasitas e microrganismos patogênicos.(AU)

Antimicrobial and insecticidal effects of essential oil and plant extracts of Myrcia oblongata DC in pathogenic bacteria and Alphitobius diaperinus / Aplicação de óleo e extratos de Myrcia oblongata DC em bactérias patogênicas e controle biológico de Alphitobius diaperinus

The secondary metabolism products of plants have influenced great economic interest, given their chemical diversity and biological activities. Because of this, this study evaluates the phytochemical composition, antimicrobial activity, insecticidal, and antioxidant activity of plant extracts and oil of Myrcia oblongata. Saponins, steroids, triterpenoids, tannins, and flavonoids were detected. The extracts showed antimicrobial capacity on the tested microorganisms, except for the methanolic extract, which showed no activity for P. mirabilis and S. enteritidis. Regarding the analysis of antioxidant compounds, the hexanic, ethyl acetate and acetone extracts showed higher antioxidant activities and also higher insecticidal performance on Alphitobius diaperinus larvae, resulting in 80% adult mortality. The results reported here show that there may be a relationship between antioxidant potential and the insecticidal effect of Myrcia oblongata DC. The components present in both the extract and the oil can be used as natural alternative to synthetic compounds in the biological control of parasites and pathogenic microorganisms.

Os produtos do metabolismo secundário das plantas têm despertado grande interesse econômico, dada sua diversidade química e atividades biológicas. Neste sentido, o estudo objetivou avaliar a composição fitoquímica, atividade antimicrobiana, inseticida e antioxidante dos extratos vegetais e óleo de Myrcia oblongata. Foram detectados a presença de saponinas, esteróides, triterpenóides, taninos e flavonóides. Os extratos apresentaram capacidade antimicrobiana sobre os microrganismos testados, exceto o extrato metanólico que não demonstrou atividade para P. mirabilis e S. Enteritidis. Quanto a análise de compostos antioxidantes observou-se que os extratos hexânico, acetato de etila e acetona apresentaram maiores atividades antioxidantes e também maior performance inseticida sobre a larva Alphitobius diaperinus e exibindo mortalidade de 80% na fase adulta. Os resultados aqui reportados mostram que pode haver uma relação entre potencial antioxidante e efeito inseticida do óleo de Myrcia oblongata; os componentes presentes tanto no extrato como o oléo podem ser utilizados como alternativa natural aos compostos sintéticos no controle biológico de parasitas e microrganismos patogênicos.

Production Layer Salmonella Enteritidis Control through Dry Fed Pre & Probiotic Products

ABSTRACT Increasing interest in multiple strain Bacillus probiotics and parietal yeast fractions as feed ingredients for egg laying hen diets has also led to food safety questions. This study was undertaken to evaluate the ability of these products to reduce Salmonella Enteritidis colonization. Sixty Hy-Line hens aged 56 weeks were placed in individual cages and fed a mash diet containing one of the following treatments, control, Bacillus spp. probiotic, yeast cell wall, or a combination of yeast cell wall and Bacillus probiotic. At 60 weeks of age all hens were challenged orally with 7 x 107 CFU/bird of Salmonella Enteritidis. At 61 weeks of age, birds were humanely euthanized, by cervical dislocation and the ceca aseptically removed and cultured for S. Enteritidis prevalence and number by the Most Probable Number method. There was no significant difference in prevalence of Salmonella Enteritidis between the control and any treatments. The control birds had 4.37 log10 MPN/g of S. Enteritidis detected in the ceca. The Probiotic group had 2.96 MPN/g, a reduction of 1.41(p 0.05) and the yeast cell wall group had 2.89 MPN/g a reduction of 1.48 (p 0.05). The combination had 3.60 MPN/g a numerical reduction of 0.78 (p=0.14). The yeast cell wall and Bacillus probiotic groups significantly reduced the amount of Salmonella Enteritidis in the ceca of the laying hens.

Epidemiological and pathological aspects of salmonellosis in cattle in southern Brazil / Aspectos epidemiológicos e patológicos de salmonelose em bovinos no Sul do Brasil

Salmonellosis is a bacterial disease that affects several domestic animal species, and is commonly diagnosed in cattle, horses, and pigs. This study aimed to describe the epidemiological and pathological findings of eleven cases of enteric salmonellosis and two cases of salmonellosis with pulmonary involvement in cattle in Rio Grande do Sul State, Brazil. Clinical signs included fever, yellow diarrhea, sometimes with blood streaks, and dyspnea, with a clinical course ranging from 1 to 30 days. Eight cases occurred as outbreaks, whereas five cases occurred individually. Risk factors included inadequate handling practices, such as overcrowded facilities and comorbidities, including anaplasmosis. The main gross finding of the enteric presentation was fibrinonecrotic enterocolitis, occasionally associated with button ulcers, mesenteric lymphadenomegaly, splenomegaly, cholecystitis and hepatomegaly. In addition, one steer with a chronic clinical progression presented severe segmental thickening of the ileum, associated with intestinal rupture and peritonitis. In the respiratory system, the main findings were reddened, non-collapsed lungs, with multifocal areas of atelectasis. The main microscopic findings were observed in the small and large intestines, and these were characterized by severe necrosis and mucosal ulceration, associated with marked inflammatory infiltrate of neutrophils and fibrin deposition intermixed by rod-shaped bacterial aggregates, and fibrosis, as well as interstitial pneumonia. Seven cases yielded positive bacterial cultures for Salmonella spp. and three serovars, namely Typhimurium, Dublin, and Panama were identified. All cases exhibited immunolabeling for Salmonella spp. using immunohistochemistry.(AU)

Salmonelose é uma doença bacteriana que afeta inúmeras espécies animais, especialmente os bovinos, os equinos e os suínos. O presente estudo teve como objetivo descrever os aspectos epidemiológicos e patológicos de onze casos de salmonelose entérica e dois de salmonelose pulmonar em bovinos no estado do Rio Grande do Sul, Brasil. Os sinais clínicos incluíram febre, diarreia amarelada, por vezes com estrias de sangue, anorexia, perda de peso e dispneia, com curso clínico que variou de um a 30 dias. Em oito casos, a doença ocorreu em forma de surtos e cinco foram individuais. Identificou-se fatores de risco relacionados ao manejo inadequado com os bovinos, como alta lotação, além de comorbidades associadas, como anaplasmose. Os principais achados macroscópicos da forma entérica consistiram em enterocolite fibrinonecrótica, por vezes associada a formação de úlceras botonosas, linfonodos mesentéricos e baço aumentados, colecistite e hepatomegalia. Ainda, um bovino com quadro clínico crônico apresentou acentuado espessamento segmentar da parede do íleo associado a ruptura intestinal e peritonite. Na forma respiratória, os principais achados incluíram pulmões não colabados, avermelhados, com áreas multifocais de atelectasia. Os principais achados microscópicos foram observados no intestino delgado e grosso e foram caracterizados por acentuada necrose e ulceração da mucosa, associada a acentuado infiltrado inflamatório de neutrófilos e deposição de fibrina entremeada por agregados bacterianos cocobacilares e fibrose, além de pneumonia intersticial. Sete casos foram positivos para Salmonella sp. no cultivo bacteriano, com identificação dos sorovares Typhimurium, Dublin e Panama. Ao exame imuno-histoquímico para Salmonella sp. todos os casos apresentaram marcação positiva nos órgãos avaliados.(AU)

Action of benzalkonium chloride in different pH

ABSTRACT Ammonium quaternary compounds are widely used in poultry and swine production as disinfectants in the control of pathogens. They act on gram-positive bacteria, gram-negative bacteria, enveloped fungi and viruses. However, in some conditions of pH and presence of organic matter can be inactivated. This study evaluated the action of ammonium quaternary compounds at 1:1,000 and 1:2,000 dilutions against Salmonella enterica serovarTyphimurium and Salmonella enterica serovar Enteritidis in the presence of three different organic matter simulators, fetal bovine serum, skim milk and whole milk concentration of 1, 3, 5, and 7% and at pH 6 and 9, with 15 min of contact. It was possible to verify that the organic matter simulators adjusted in the same conditions of contact time and percentage, in the in vitro tests, presented different results and the fetal bovine serum did not inactivate the disinfectant. However, the best result against S. Typhimurium and S. Enteritidis was obtained at pH 6 at the dilution of 1:1,000 in all organic matter simulators.

Action of benzalkonium chloride in different pH

Ammonium quaternary compounds are widely used in poultry and swine production as disinfectants in the control of pathogens. They act on gram-positive bacteria, gram-negative bacteria, enveloped fungi and viruses. However, in some conditions of pH and presence of organic matter can be inactivated. This study evaluated the action of ammonium quaternary compounds at 1:1,000 and 1:2,000 dilutions against Salmonella enterica serovarTyphimurium and Salmonella enterica serovar Enteritidis in the presence of three different organic matter simulators, fetal bovine serum, skim milk and whole milk concentration of 1, 3, 5, and 7% and at pH 6 and 9, with 15 min of contact. It was possible to verify that the organic matter simulators adjusted in the same conditions of contact time and percentage, in the in vitro tests, presented different results and the fetal bovine serum did not inactivate the disinfectant. However, the best result against S. Typhimurium and S. Enteritidis was obtained at pH 6 at the dilution of 1:1,000 in all organic matter simulators.

Validation of Alternative Methods of Detection of Salmonella Spp. in Experimentally Contaminated Poultry Environmental Samples

The analysis of Salmonella in the feces and the birds environment is a way of monitoring the colonization in the flocks and verifying the need for the introduction of stricter controls, in such a way that the results of the tests should be known before being sent for slaughter. The polymerase chain reaction (PCR), as well as other rapid methods represent alternatives increasingly used to detect enteric pathogens, but they need proof of effectiveness for their wide use. The aim of this study was to evaluate the equivalence between the results obtained by the methods: real-time PCR (BAX® System), Modified Rappaport-Vassiliadis Semi-solid Medium (MSRV) (ISO 6579) and the traditional method of official reference in Brazil for research of S. Typhimurium and S. Enteritidis in poultry samples. Two hundred and fifty-two samples of disposable shoe covers (DSC) and 252 samples of feces were infected with an average of 2 to 3 log CFU/g of each serovar, and the same samples without fortification were evaluated by the three methods. Five hundred and four diagnoses were obtained with satisfactory results in terms of repeatability (greater than 80%), reproducibility (mean 83,1%), sensitivity (81% to 100%), specificity (95% to 100%), and accuracy (90% to 100%). The compliance test verified that there was not a significant difference between the alternative and the official methods, allowing us to state that the methodologies have had equivalent performances.(AU)

Validation of Alternative Methods of Detection of Salmonella Spp. in Experimentally Contaminated Poultry Environmental Samples

The analysis of Salmonella in the feces and the birds environment is a way of monitoring the colonization in the flocks and verifying the need for the introduction of stricter controls, in such a way that the results of the tests should be known before being sent for slaughter. The polymerase chain reaction (PCR), as well as other rapid methods represent alternatives increasingly used to detect enteric pathogens, but they need proof of effectiveness for their wide use. The aim of this study was to evaluate the equivalence between the results obtained by the methods: real-time PCR (BAX® System), Modified Rappaport-Vassiliadis Semi-solid Medium (MSRV) (ISO 6579) and the traditional method of official reference in Brazil for research of S. Typhimurium and S. Enteritidis in poultry samples. Two hundred and fifty-two samples of disposable shoe covers (DSC) and 252 samples of feces were infected with an average of 2 to 3 log CFU/g of each serovar, and the same samples without fortification were evaluated by the three methods. Five hundred and four diagnoses were obtained with satisfactory results in terms of repeatability (greater than 80%), reproducibility (mean 83,1%), sensitivity (81% to 100%), specificity (95% to 100%), and accuracy (90% to 100%). The compliance test verified that there was not a significant difference between the alternative and the official methods, allowing us to state that the methodologies have had equivalent performances.

Antibacterial activity of chitosan biofilm for the conservation of fertile and table eggs / Atividade antibacteriana de biofilme de quitosana para conservação de ovos férteis e de mesa

The aim of this study was to develop a chitosan biofilm against Salmonella enteritidis, for the conservation of fertile and table eggs. Two experiments were performed. Experiment 1: 400 specific pathogen-free table eggs were divided in a completely randomized design into four treatments, five replicates and each replicate with 20 table eggs. Experimental groups were assigned to control and 1, 5 and 10% chitosan treatment. The eggs were immersed in the chitosan solution. They were then exposed to Salmonella enteritidis and stored for 1, 24, 96 and 168h at 4ºC. The eggs were then washed with 10mL of physiological saline solution. Experiment 2: 80 specific pathogen-free fertile eggs were tested, the assays were assigned to control and 1, 5 and 10% chitosan treatment. Each treatment had 20 fertile eggs. The eggs were immersed in the chitosan solution. They were individually weighed and incubated. Egg weight, humidity loss, and hatchability (weight and length of newly hatched chicks) characteristics were assessed. In Experiment 1, comparison between treatments showed differences (P< 0.05) in the total recovered of Salmonella enteritidis on eggshell, with the lower values in 5 y 10% chitosan treatment at 96 y 168h respectively. In Experiment 2, chitosan did not show any effect on the egg weight and chick weight, where the average was 57.44 and 38.23g respectively. The humidity loss and chick length showed differences (P< 0.05), with the lower values in 5 y 10% chitosan treatment. The antibacterial activity of chitosan biofilm provide a practical tool against Salmonella enteritidis in fertile and table eggs because the chitosan did not affect egg weight and chick weight, relevant parameters in the poultry industry.(AU)

O presente estudo teve como objetivo desenvolver um biofilme de quitosana contra Salmonella enteritidis, para conservação de ovos férteis e de mesa. Dois experimentos foram realizados. Experimento 1: 400 ovos de mesa livres de patógenos especificados foram divididos em delineamento inteiramente casualizado em quatro tratamentos, cinco repetições e cada réplica contendo 20 ovos de mesa. Grupos experimentais foram designados para controle e 1, 5 e 10% de tratamento com quitosana. Os ovos foram imersos em solução de quitosana. Em seguida foram expostos a Salmonella enteritidis, e armazenados por 1, 24, 96 e 168h a 4ºC. Após, os ovos foram lavados com 10mL de solução salina fisiológica. Experimento 2: 80 ovos férteis livres de patógenos especificados foram testados. Os ensaios foram atribuídos a controle e 1, 5 e 10% de tratamento com quitosana. Cada tratamento teve 20 ovos férteis. Os ovos foram imersos em solução de quitosana. Em seguida foram individualmente pesados e incubados. Peso dos ovos, perda de umidade e características de eclodibilidade (peso e comprimento dos pintinhos recém-nascidos) foram avaliados. No Experimento 1, a comparação entre tratamentos mostrou diferenças (P< 0,05) na quantidade total recuperada de Salmonella enteritidis na casca, com os menores valores em 5 e 10% de tratamento com quitosana a 96 e 168h respetivamente. No experimento 2, a quitosana não mostrou nenhum efeito no peso do ovo e no peso do pintinho, onde a média foi de 57,44 e 38,23g respetivamente. A perda de umidade e comprimento do pintinho apresentaram diferenças (P< 0,05), com os menores valores em 5 e 10% de tratamento com quitosana. A atividade antibacteriana do biofilme de quitosana, fornece uma ferramenta prática contra Salmonella enteritidis em ovos férteis e de mesa, pois a quitosana não afetou o peso do ovo e peso do pintinho, parâmetros relevantes na indústria avícola.(AU)

Antibacterial activity of chitosan biofilm for the conservation of fertile and table eggs / Atividade antibacteriana de biofilme de quitosana para conservação de ovos férteis e de mesa

The aim of this study was to develop a chitosan biofilm against Salmonella enteritidis, for the conservation of fertile and table eggs. Two experiments were performed. Experiment 1: 400 specific pathogen-free table eggs were divided in a completely randomized design into four treatments, five replicates and each replicate with 20 table eggs. Experimental groups were assigned to control and 1, 5 and 10% chitosan treatment. The eggs were immersed in the chitosan solution. They were then exposed to Salmonella enteritidis and stored for 1, 24, 96 and 168h at 4ºC. The eggs were then washed with 10mL of physiological saline solution. Experiment 2: 80 specific pathogen-free fertile eggs were tested, the assays were assigned to control and 1, 5 and 10% chitosan treatment. Each treatment had 20 fertile eggs. The eggs were immersed in the chitosan solution. They were individually weighed and incubated. Egg weight, humidity loss, and hatchability (weight and length of newly hatched chicks) characteristics were assessed. In Experiment 1, comparison between treatments showed differences (P< 0.05) in the total recovered of Salmonella enteritidis on eggshell, with the lower values in 5 y 10% chitosan treatment at 96 y 168h respectively. In Experiment 2, chitosan did not show any effect on the egg weight and chick weight, where the average was 57.44 and 38.23g respectively. The humidity loss and chick length showed differences (P< 0.05), with the lower values in 5 y 10% chitosan treatment. The antibacterial activity of chitosan biofilm provide a practical tool against Salmonella enteritidis in fertile and table eggs because the chitosan did not affect egg weight and chick weight, relevant parameters in the poultry industry.(AU)

O presente estudo teve como objetivo desenvolver um biofilme de quitosana contra Salmonella enteritidis, para conservação de ovos férteis e de mesa. Dois experimentos foram realizados. Experimento 1: 400 ovos de mesa livres de patógenos especificados foram divididos em delineamento inteiramente casualizado em quatro tratamentos, cinco repetições e cada réplica contendo 20 ovos de mesa. Grupos experimentais foram designados para controle e 1, 5 e 10% de tratamento com quitosana. Os ovos foram imersos em solução de quitosana. Em seguida foram expostos a Salmonella enteritidis, e armazenados por 1, 24, 96 e 168h a 4ºC. Após, os ovos foram lavados com 10mL de solução salina fisiológica. Experimento 2: 80 ovos férteis livres de patógenos especificados foram testados. Os ensaios foram atribuídos a controle e 1, 5 e 10% de tratamento com quitosana. Cada tratamento teve 20 ovos férteis. Os ovos foram imersos em solução de quitosana. Em seguida foram individualmente pesados e incubados. Peso dos ovos, perda de umidade e características de eclodibilidade (peso e comprimento dos pintinhos recém-nascidos) foram avaliados. No Experimento 1, a comparação entre tratamentos mostrou diferenças (P< 0,05) na quantidade total recuperada de Salmonella enteritidis na casca, com os menores valores em 5 e 10% de tratamento com quitosana a 96 e 168h respetivamente. No experimento 2, a quitosana não mostrou nenhum efeito no peso do ovo e no peso do pintinho, onde a média foi de 57,44 e 38,23g respetivamente. A perda de umidade e comprimento do pintinho apresentaram diferenças (P< 0,05), com os menores valores em 5 e 10% de tratamento com quitosana. A atividade antibacteriana do biofilme de quitosana, fornece uma ferramenta prática contra Salmonella enteritidis em ovos férteis e de mesa, pois a quitosana não afetou o peso do ovo e peso do pintinho, parâmetros relevantes na indústria avícola.(AU)

Evaluation of propanediol and cobalamin metabolism in the intestinal colonization and systemic invasion of Salmonella Enteritidis in laying hens / [Avaliação do metabolismo do propanodiol e da cobalamina na colonização intestinal e na infecção sistêmica de Salmonella Enteritidis em poedeiras]

Embora Salmonella Enteritidis (SE) seja capaz de metabolizar 1,2-propanodiol (1,2-Pd), utilizado como fonte de carbono e de energia ao longo de uma rota dependente de vitamina B12, a importância deste composto na infeção de Gallus gallus domesticus por SE permanece desconhecida. No presente estudo, foram construídos um mutante de SE sem os genes pduCDE, que codifica a propanodiol desidratase (Pdu), e outro contendo as deleções no pduCDE e também nos genes cobS e cbiA, responsáveis pela síntese de vitamina B12. Em seguida, avaliou-se a importância do metabolismo do 1,2-Pd em SE para colonização intestinal de infecção sistêmica de poedeiras comerciais. As estirpes mutantes de SE foram capazes de colonizar o intestino, de serem excretadas nas fezes e de invadir o baço e o fígado na mesma intensidade que a estirpe selvagem, o que sugere que os produtos dos genes pduC, pduD, pduE, cobS e cbiA não são essenciais durante infecção por Salmonella Enteritidis nessa espécie.(AU)

Effects of Glutamine on the Mucosal Structure and Immune Cells in the Intestines of Broiler Chickens Challenged with Salmonella Enteritidis

The aim of this study was to investigate the effects of glutamine (Gln) on the intestinal mucosal structure and immune cells of broilers infected with Salmonella Enteritidis. 160 1-d-old commercial Arbor Acres (AA) broilers were randomly selected to receive one of four treatments, each of which had 5 replicates. Each replicate consisted of 8 chicks subjected to a 21-d feeding trial. Group I served as the unchallenged (CON). All birds in groups II (SCC) - IV were challenged with 2.0 × 104 CFU/mL of S. Enteritidis. The birds in groups III and IV were treated with 0.5% and 1.0% Gln. The results showed that S. Enteritidis infection led to a decrease in the relative length and weight, villus height:crypt depth (VH:CD) of the jejunum and ileum, the number of intraepithelial lymphocyte cells, and number of goblet cells and an increase in the number of mast goblet cells compared with the measurements of these parameters in the CON group (p 0.05). In addition, the Gln groups had increased relative length and weight, VH:CD of the jejunum and ileum, numbers of intraepithelial lymphocyte cells, and numbers of goblet cells and decreased crypt depth in the jejunum and ileum and numbers of mast goblet cells compared with the measurements of these parameters in the SCC group (p 0.05). It was concluded that Gln added to broiler diets can effectively alleviate the intestinal mucosal damage caused by S. Enteritidis infection and improve its normal defense barrier function.(AU)

Effects of Glutamine on the Mucosal Structure and Immune Cells in the Intestines of Broiler Chickens Challenged with Salmonella Enteritidis

The aim of this study was to investigate the effects of glutamine (Gln) on the intestinal mucosal structure and immune cells of broilers infected with Salmonella Enteritidis. 160 1-d-old commercial Arbor Acres (AA) broilers were randomly selected to receive one of four treatments, each of which had 5 replicates. Each replicate consisted of 8 chicks subjected to a 21-d feeding trial. Group I served as the unchallenged (CON). All birds in groups II (SCC) - IV were challenged with 2.0 × 104 CFU/mL of S. Enteritidis. The birds in groups III and IV were treated with 0.5% and 1.0% Gln. The results showed that S. Enteritidis infection led to a decrease in the relative length and weight, villus height:crypt depth (VH:CD) of the jejunum and ileum, the number of intraepithelial lymphocyte cells, and number of goblet cells and an increase in the number of mast goblet cells compared with the measurements of these parameters in the CON group (p 0.05). In addition, the Gln groups had increased relative length and weight, VH:CD of the jejunum and ileum, numbers of intraepithelial lymphocyte cells, and numbers of goblet cells and decreased crypt depth in the jejunum and ileum and numbers of mast goblet cells compared with the measurements of these parameters in the SCC group (p 0.05). It was concluded that Gln added to broiler diets can effectively alleviate the intestinal mucosal damage caused by S. Enteritidis infection and improve its normal defense barrier function.

Efficiency of chitosan synergism with clove essential oil in the coating of intentionally contaminated Tambaqui fillets / Eficiência do sinergismo de quitosana com óleo essencial de cravo no revestimento de filés de tambaqui contaminados intencionalmente

The edible coating of chitosan with clove essential oil (CEO) was studied for its ability to reduce the microbial growth of pathogens (Escherichia coli O157:H7 CDCEDL933, Listeria monocytogenes CERELA, Salmonella Enteritidis ATCC13076, Staphylococcus aureus ATCC43300, and Pseudomonas aeruginosa ATCC27853) in Tambaqui fillets kept under refrigeration. In in vitro tests, chitosan showed higher antimicrobial activity against S. aureus and L. monocytogenes (MIC 0.5%), and CEO for L. monocytogenes (MIC 0.08%). Based on the antimicrobial activity of chitosan and CEO, Tambaqui fillets were subjected to different treatments, T1: chitosan 2%; T2: chitosan 2% + CEO 0.16%, and T3: chitosan 0.5% + CEO 0.08%, kept at 4 ºC for 72 h. The chitosan coating, incorporated with CEO, inhibited microorganisms in Tambaqui fillets and enhanced coating efficiency (p < 0.05). It was most effective against L. monocytogenes and S. aureus at the lowest CEO concentration (0.08%). Chitosan coating in combination with CEO enhanced the antimicrobial effect of pathogens on Tambaqui fillets, increased their shelf life under refrigeration, and was more effective against Gram-positive pathogens than Gram-negative pathogens.(AU)

O revestimento comestível de quitosana com óleo essencial de cravo (OEC) foi estudado por sua capacidade em reduzir o crescimento microbiano de patógenos (Escherichia coli O157:H7 CDCEDL933, Listeria monocytogenes CERELA Salmonella Enteritidis ATCC 13076, Staphylococcus aureus ATCC43300 e Pseudomonas aeruginosa ATCC27853) em filés de tambaqui mantidos sob refrigeração. Nos testes in vitro, a quitosana apresentou maior atividade antimicrobiana para S. aureus e L. monocytogenes (CIM 0,5%) e o OEC para L. monocytogenes (CIM 0,08%). Com base na atividade antimicrobiana da quitosana e OEC, os filés de Tambaqui foram submetidos a T1: quitosana a 2%; T2: quitosana 2% + OEC 0,16% e T3: quitosana 0,5% + OEC 0,08%, mantidos a 4 ºC por 72 h. O revestimento de quitosana, incorporado ao OEC, inibiu os micro-organismos nos filés de Tambaqui aumentando a eficiência do revestimento (p< 0,05); e foi mais eficaz para L. monocytogenes e S. aureus na menor concentração do OEC (0,08%). O revestimento de quitosana quando combinado ao OEC aumentou o efeito antimicrobiano de patógenos nos filés de Tambaqui, aumentando sua vida útil sob refrigeração, sendo mais eficaz contra patógenos Gram positivos do que os patógenos Gram negativos.(AU)