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)

An outbreak of fatal Pullorum disease (Salmonella Pullorum) in Guinea fowl keets (Numida meleagris) / Surto fatal de pulorose (Salmonella Pullorum) em pintinhos de galinhas-d'Angola (Numida meleagris)

Pullorum disease is described worldwide and is caused by Salmonella enterica subspecies enterica serovar Gallinarum biovar Pullorum (S. Pullorum). S. Pullorum infection is important in commercial poultry, provoking a systemic disease with high mortality rates. Its occurrence requires notification, and when it is diagnosed in commercial breeding flocks, its eradication is demanded. The aim of this study was to report a severe outbreak of Pullorum disease in young Guinea fowl (Numida meleagris), resulting in 100% mortality of keets (n=290) within the first two weeks of age. All examined keets had enlarged liver, kidneys and spleen (5/5), and the affected tissues were submitted to histological and bacteriological examination. On histopathology, random paratyphoid nodules characterized by areas of necrosis with fibrin and a moderate infiltrate of macrophages and heterophils were observed in the liver. In kidneys, discrete areas of necrosis associated with moderate multifocal infiltrates of lymphocytes, and plasma cells were observed. In the spleen, a moderate infiltrate of macrophages was noticed. Isolation of colonies suggestive of S. Pullorum from liver and spleen was performed in selective agars and, after biochemical tests, confirmed by specific duplex-PCR. The antimicrobial susceptibility test of the isolated strain revealed resistance to only sulfamethoxazole + trimethoprim among the tested antimicrobials. The S. Pullorum isolate recovered in the present study was highly pathogenic to N. meleagris and may represent a risk to other avian species, including industrial poultry.

A pulorose é descrita mundialmente e é causada por Salmonella enterica subespécie enterica sorovar Gallinarum biovar Pullorum (S. Pullorum). A infecção por S. Pullorum é importante em aves comerciais, provocando doença sistêmica com altas taxas de mortalidade. Sua ocorrência requer notificação e quando diagnosticada em aves de criação comercial resulta na erradicação do plantel. O objetivo deste estudo foi relatar um surto grave de pulorose em filhotes de galinhas-d'Angola (Numida meleagris), resultando em 100% de mortalidade das aves (n=290) nas primeiras duas semanas de idade. Os pintinhos recebidos tinham hepato, espleno e nefromegalia (5/5). Os tecidos dos cinco indivíduos recebidos foram submetidos a exame histológico e bacteriológico. Na histopatologia, foram observados nódulos paratifoides aleatórios caracterizados por áreas de necrose com fibrina e infiltrado moderado de macrófagos e heterófilos no fígado. Nos rins, foram observadas áreas discretas de necrose associadas a infiltrados multifocais moderados de linfócitos e plasmócitos. No baço, foi observado infiltrado moderado de macrófagos. O isolamento de colônias sugestivas de S. Pullorum de fígados e baços foi realizado em ágares seletivos e, após testes bioquímicos, confirmado por duplex-PCR específico. A susceptibilidade antimicrobiana da cepa isolada revelou resistência apenas ao sulfametoxazol + trimetoprim entre os antimicrobianos testados. O isolado de S. Pullorum recuperado no presente estudo foi altamente patogênico para N. meleagris e pode representar um risco para outras espécies de aves, incluindo aves industriais.

Prevalence and antimicrobial resistance of Salmonella spp isolated from free-ranging wild boars in the state of São Paulo, Brazil / Prevalência e resistência antimicrobiana de Salmonella spp isoladas em javalis de vida livre no estado de São Paulo

European wild boars (Sus scrofa) are considered exotic invasive species worldwide. Invasions of wild boars are a growing public health concern, as wild boars may represent an important reservoir of zoonotic pathogens, including bacteria of the genus Salmonella. The aim of this study was to determine the prevalence and serovars of Salmonella spp. in free-ranging wild boars legally hunted in the state of São Paulo, Brazil, and the susceptibility of those Salmonella spp. to antimicrobials. Fecal samples and mesenteric lymph nodes were acquired from 63 wild boars. The prevalence of Salmonella spp. in free-ranging wild boars was 9.5 % (6/63; confidence interval: 4.4 % - 19.2 %). Six serovars were isolated: S. enterica subsp. enterica ser. 4,5,12:-:1,2, S. enterica ser. Cerro, S. enterica ser. Madelia, S. enterica ser. Typhimurium, S. enterica ser. I (4,5,12:i:-) and S. enterica ser. Muenster. Analysis of antimicrobial resistance of Salmonella spp. showed that the majority of serovars were fully susceptible to the tested antimicrobials. Only S. enterica ser. Typhimurium and S. enterica ser. Muenster showed a resistance pattern to at least one antimicrobial analyzed. To our knowledge, this study is the first report the prevalence and serovars of Salmonella spp. in free-ranging wild boars in the State of São Paulo, Brazil. Results indicate a low prevalence with variability of Salmonella serovars, with some pattern of antimicrobial resistance. This study highlights the potential role of wild boars as carriers of Salmonella and could pose a risk to wild and domestic animals as well as humans.

Os javalis europeus (Sus scrofa) são considerados uma espécie exótica invasora em todo o mundo. As invasões de javalis são uma preocupação crescente de saúde pública, pois os javalis podem representar um importante reservatório de patógenos zoonóticos, incluindo bactérias do gênero Salmonella. O objetivo deste estudo foi determinar a prevalência e os sorovares de Salmonella spp. em javalis de vida livre caçados legalmente no Estado de São Paulo, Brasil, e a suscetibilidade dessa Salmonella spp. aos antimicrobianos. Amostras fecais e linfonodos mesentéricos foram adquiridos de 63 javalis. A prevalência de Salmonella spp. em javalis selvagens foi de 9,5% (6/63; intervalo de confiança: 4,4% - 19,2%). Seis sorovares foram isolados: S. enterica subsp. enterica ser. 4,5,12:-:1,2, S. enterica ser. Cerro, S. enterica ser. Madelia, S. enterica ser. Typhimurium, S. enterica ser. I (4,5,12:i:-) e S. enterica ser. Muenster. As análises de resistência antimicrobiana de Salmonella spp. evidenciaram que a maioria dos sorovares era pansensível aos antimicrobianos testados. Apenas S. enterica ser. Typhimurium e S. enterica ser. Muenster mostraram um padrão de resistência a pelo menos um antimicrobiano analisado. A saber, este estudo é o primeiro relato da prevalência e de sorovares de Salmonella spp. em javalis de vida livre no Estado de São Paulo, Brasil. Os resultados indicaram baixa prevalência com variabilidade de sorovares de Salmonella, com algum padrão de resistência antimicrobiana. Este estudo destaca o papel potencial dos javalis como portadores de Salmonella spp. e pode representar um risco para os animais domésticos e selvagens, bem como para os humanos.

The Efficiency of Bacteriophages Against Salmonella Typhimurium Infection in Native Noi Broilers

Though recently considered a therapeutic treatment for commercial broilers, little is known about the effects of bacteriophages on native, slow-growing birds. This study evaluated their efficacy against Salmonella enterica subsp. enterica serovar Typhimurium infected Noi chicken, a native Vietnamese broiler breed. In total, 420 birds were used in a completely randomized design consisting of seven treatments and four replicates of 15 birds. The treatments were NC (negative control), PC (positive control, S. Typhimurium challenged); NC+B1 and NC+B2 (negative control plus B1 or B2 bacteriophage, respectively); PC+B1, PC+B2 (positive control plus B1 or B2 bacteriophage, respectively) and PC+B1B2 (positive control plus both B1 and B2 bacteriophages). After four weeks of infection, the mortality rate in the PC group was 51.1% compared with 11.1% in the PC+B1B2 treatment. Bacteriophage administration had resulted in increased weight gain and decreased feed conversion ratio, particularly when both phages were included in the treatment (p<0.001). Moreover, the relative percentage of carcass weight was lowest in the PC treatment (66.9%) (p<0.001), whereas the other treatments registered similar carcass weight values. Regarding the internal organs, liver weight percentage was higher in the non-treated Salmonella group, and enlarged spleens were also noted in infected chickens even when treated with bacteriophages. The correlation between phage administration and blood parameters was unclear. Although the use of two bacteriophages for therapy was determined to be preferable for the majority of the criteria examined, further genetic characterization of the phages will be required before they can be widely used in chicken farms.(AU)

Salmonella Pullorum em criação de galinhas caipiras no Oeste da Bahia, Brasil / Salmonella Pullorum in a free-range chicken farm in Western Bahia, Brazil

Background: Industrial poultry farming has developed progressively in Brazil, conferring the country a prominent position on the national and international scene. Likewise, alternative poultry farming is an important economic activity for small-scale family farmers. However, shortcomings related to sanitary management lead to increased occurrence of avian diseases, such as those caused by Salmonella spp. Despite salmonellosis has been described in industrial establishments, reports in alternative farms are less common, therefore the objective of this study was to describe the occurrence of salmonellosis in free-range chickens in the municipality of Barra, Western Bahia, Brazil. Cases: The poultry farmer reported the occurrence of diarrhoea in his chicken flock since the acquisition of the batch of chicks. Initially, carrying out the medicinal treatment of the birds, there was clinical improvement, however, successive recurrences of clinical signs occurred, such as diarrhoea, apathy, anorexia, and death of some birds. Upon learning about the case, an epidemiological investigation of the flock was carried out, and it was noted that some of the birds were retracted, apathetic, anorexic, and a lot of diarrheic faeces of a yellowish-white appearance were also observed. The entire flock had a history of vaccination against diseases: newcastle disease, infectious bronchitis, gumboro disease, and fowlpox. For better evaluation, five birds were necropsied, enabling the observation that the animals had a good body score. However, the necropsy revealed lesions such as splenomegaly, hepatomegaly, and enteritis in the three birds initially analysed (Animals 1, 2, and 3). The other birds (Animals 4 and 5) were submitted to evaluation for Eimeria sp. oocysts by means of scrapings from the intestinal mucosa, and there were no structures compatible with oocysts. Faecal samples were collected from another six birds in the flock for coproparasitological examination, and the presence of oocysts was not detected. Finally, sera from 20 birds in the flock were collected for the Rapid Serum Agglutination Test (SAR) for the detection anti-Salmonella Pullorum antibodies. Discussion: The diagnosis was based on clinical evaluation, post mortem pathological findings of the necropsied birds, epidemiological data and confirmed with SAR testing, whereby 11 birds were seropositive for Salmonella enterica subspecies enterica serovar Pullorum. The prevalence of S. Pullorum is poorly described in alternative farming. Based on the farmer's report, it is believed that the chicks were purchased already infected, because birds from the same batch, also purchased by neighbouring producers, showed the same clinical signs. The unsatisfactory sanitation in the flock was another factor that may have favoured the persistence of the bacteria, since, the lack of removal of organic matter is a source of nutrients for microorganisms, and this may have favoured the multiplication and maintenance of the bacteria in the down feathers, feed, and water. The elimination of S. Pullorum through the faeces, in addition to the density of the birds, may have led to transmission to the other healthy birds. However, the knowledge and adoption of prophylactic measures in free-range chicken farms is a crucial factor in minimizing the occurrence of outbreaks and thus avoiding a future public Health Problem.

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

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)

Salmonellosis in a young Amazonian manatee (Trichechus inunguis) / Salmonelose em um filhote de peixe-boi-da-amazônia (Trichechus inunguis)

The objective of this study was to describe a case of salmonellosis in an Amazonian manatee (Trichechus inunguis). The animal was rescued from a tributary of the Capim River, in the municipality of Goianésia do Pará, Pará, Brazil, kept in a pool, and died approximately five months after being rescued. The main changes observed at necropsy were that the cecum and colon had serosal hyperemia, wall edema, thickened mucosa with ulcerated areas and covered by a thin layer of fibrin, and a yellowish liquid content. Histologically, there was moderate multifocal fibrinonecrotic typhlocolitis associated with intense bacterial colonization. Salmonella enterica subsp. enterica (rough) was isolated in cecum and colon fragments. This is the first report of salmonellosis in an Amazonian manatee.(AU)

O objetivo deste trabalho é descrever um caso de salmonelose em um peixe-boi-da-amazônia (Trichechus inunguis). O animal foi resgatado em um afluente do rio Capim, no município de Goianésia do Pará, Pará, Brasil, sendo mantido em uma piscina e morrendo aproximadamente cinco meses após o resgate. As principais alterações observadas na necropsia foram ceco e colón com hiperemia da serosa; edema da parede; mucosa espessada, com áreas ulceradas e revestida por uma fina camada de fibrina; e conteúdo líquido amarelado. Histologicamente havia tiflocolite fibrinonecrótica, multifocal, moderada, associada a intensa colonização bacteriana. Em fragmentos de ceco e cólon foi isolada Salmonella enterica subsp. enterica (rugosa). Este é o primeiro relato de salmonelose em peixe-boi-da-amazônia.(AU)

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)

Association between non-typhoidal Salmonella isolated from commercial poultry sheds and associated factors in Paraná, Brazil: Cross-sectional retrospective study

The epidemiology of salmonellosis in poultry is complex, which makes it difficult to identify the origin and spread of this disease in poultry farms. The aims of this study were to characterize the spatial distribution of Salmonella enterica in epidemiological units in Paraná, Brazil; and to investigate correlations between this microorganism and associated factors. Among the epidemiological units, 78 of 243 (32.10%) were positive. Spatially, the northwestern and western regions had higher concentrations of positive cases than the other regions. In bivariate analyses, the presence of other animal species in the epidemiological unit (prevalence ratio, PR = 0.64; 95% confidence interval, CI = 0.43­0.95; p = 0.022) and proximity to establishments at risk (PR = 0.51; 95% CI = 0.32­0.81; p = 0.001) did not influence positivity, but the average population per poultry shed (between 30,501 and 32,500; PR = 2.57; 95% CI = 1.72­3.83; p = 0.001) was associated with Salmonella positivity. Multiple logistic regression demonstrated that the average population per poultry shed, presence of surrounding risk-posing establishments and presence of surrounding poultry sheds produced a significant multiple model for S. enterica. The results indicated that the presence of S. enterica may be related to higher density broiler in poultry sheds, presence of surrounding poultry sheds, proximity between positive and negative epidemiological units and altitude of the municipality. The information obtained showed that some factors were related to positivity for this microorganism and emphasizes the importance of serotyping to obtain other epidemiological data.

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.

Association between non-typhoidal Salmonella isolated from commercial poultry sheds and associated factors in Paraná, Brazil: Cross-sectional retrospective study

ABSTRACT The epidemiology of salmonellosis in poultry is complex, which makes it difficult to identify the origin and spread of this disease in poultry farms. The aims of this study were to characterize the spatial distribution of Salmonella enterica in epidemiological units in Paraná, Brazil; and to investigate correlations between this microorganism and associated factors. Among the epidemiological units, 78 of 243 (32.10%) were positive. Spatially, the northwestern and western regions had higher concentrations of positive cases than the other regions. In bivariate analyses, the presence of other animal species in the epidemiological unit (prevalence ratio, PR = 0.64; 95% confidence interval, CI = 0.430.95; p = 0.022) and proximity to establishments at risk (PR = 0.51; 95% CI = 0.320.81; p = 0.001) did not influence positivity, but the average population per poultry shed (between 30,501 and 32,500; PR = 2.57; 95% CI = 1.723.83; p = 0.001) was associated with Salmonella positivity. Multiple logistic regression demonstrated that the average population per poultry shed, presence of surrounding risk-posing establishments and presence of surrounding poultry sheds produced a significant multiple model for S. enterica. The results indicated that the presence of S. enterica may be related to higher density broiler in poultry sheds, presence of surrounding poultry sheds, proximity between positive and negative epidemiological units and altitude of the municipality. The information obtained showed that some factors were related to positivity for this microorganism and emphasizes the importance of serotyping to obtain other epidemiological data.

Salmonelose em bezerros pelo sorovar Dublin no estado do Paraná, Brasil - aspectos clínico-patológicos / Salmonellosis in calves by serovar Dublin in Paraná State, Brazil - clinicopathological aspects

Background: Calf diarrhea remains one of the main diseases affecting the cattle industry. Persistence of this significant problem is associated with the complexity of factors that may be involved (infectious, environmental). An accurate diagnosis is essential for confirming the cause and helping clinicians and cattle producers to apply appropriate strategies in a timely manner. This report describes the histological changes according to the degree of salmonellosis severity, which is a contagious infectious disease caused by Salmonella enterica subsp. enterica, a Gram-negative bacterium, in two beef calves in northern Paraná State, southern Brazil. Cases: Two 90-day-old crossbred Angus and Nellore calves from a beef farm in northern Paraná State were referred to a Veterinary Hospital. Animal 1- developed acute clinical signs (enteritis, ataxia, and muscle rigidity) and died the day after the signs began. Gross findings included heavy and non-collapsed lungs, pulmonary oedema, hepatomegaly, enteritis, and severe diffuse typhlitis. Microscopic analysis revealed severe diffuse necrotic enteritis, typhlitis, severe diffuse interstitial pneumonia, moderate centrilobular hepatic necrosis, mild multifocal nephritis, and severe spleen and lymph node necrosis. Paratyphoid nodules were evidenced on the liver, spleen, and mesenteric lymph nodes. Animal 2- presented apathy, green fibrinous diarrhea, and died three days after the onset of clinical signs. The macroscopic examination showed moderate diffuse enteritis and hepatosplenomegaly. At the microscopic examination, many paratyphoid nodules were observed on the liver, spleen, kidneys, and Peyer's patches, which were associated with intralesional and intravascular bacilli. Additional findings included severe diffuse fibrinous necrotic enteritis with intralesional bacilli, diffuse interstitial pneumonia, multifocal splenic necrosis, lymphoid depletion, and moderate multifocal to coalescent lymphocytic nephritis. Histological Gram staining was performed on selected samples, revealing intralesional Gram-negative bacilli in the liver and intestine. Thus, liver and intestine fragments were used for the microbiological examination. Microbiological culture, isolation and biochemical tests identified the genus Salmonella spp. Then, the colonies were subjected to serological tests for serovar identification, according to ISO/TR 6579-3, which determined the serovar Dublin. After identifying the disease etiological agent, the outbreak was controlled by appropriate antibiotic therapy combined with the correction of sanitary measures. Discussion: Enteritis is a frequent disease in calves, posing a diagnostic challenge in identifying the etiological agent. In the present case, the histological, microbiological, and serological results confirmed a disseminated Salmonella spp. infection. The microscopic findings, such as interstitial pneumonia, fibrinous necrotic enteritis markedly in ileum and paratyphoid nodules in various organs, are the most common aspects of the disease. However, fibrinous cholecystitis, which is considered pathognomonic for salmonellosis, was not observed in this study. Therefore, the absence of such a lesion should not exclude the disease in sick animals. A microscopic injury score was used to determine lesion severity by assigning values from 1 to 4, wherein: 1 = no apparent lesions, 2 = mild lesions, 3 = moderate lesions, and 4 = severe lesions. Both calves were scored as 4. Multiple predisposing factors for the condition were identified in this farm such as different age animals in the same paddock and no specific paddock for sick animals, given that the infectious agent remains in feces, saliva, and nasal discharge. The serovar Dublin induces several clinical signs such as septicemic, respiratory, and enteric manifestations, making a clinical diagnosis a challenge

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.

Plasmid-Mediated Antibiotic Resistance and Class 1 Integron in Salmonella Heidelberg Isolated from Poultry Farms in Santander - Colombia

Salmonella enterica is a zoonotic pathogen transmitted to humans by the consumption of contaminated poultry products. Salmonella causes around 93 million cases of gastroenteritis and 155,000 deaths worldwide. A high number of multidrug-resistant Salmonella isolates has been found from different segments of poultry production, and it is often associated with horizontal antibiotic resistance gene transfer through mobile elements such as plasmids, integrons, and transposons. The emergence of antibiotic-resistant Salmonella has been associated with the misuse of antibiotics in animal production and it is classified as a highly important pathogen from public health due to its zoonotic character and its high dissemination ability. This study aimed to determine the antibiotic resistance associated with plasmids, and class 1 and 2 integrons from Salmonella Heidelberg isolates from healthy chickens in poultry farms from Santander, Colombia. 15 Salmonella Heidelberg isolates obtained from cloacal samples, were evaluated through endpoint PCR to determine the presence of plasmid-encoded resistance genes, and class 1 and 2 integrons. The bla CMY2 , strA and strB, sul1 and sul2, and tetB genes were found in all 15 Salmonella Heidelberg. The class 1 integron was present in 11 out of 15 isolates, harbored the gene cassette dfra7.The results indicate the presence of a high number of antibiotic resistance genes associated with plasmids and class 1 integrons in Salmonella Heidelberg strains isolated from poultry farms, resulting in a public health concern, both in humans and poultry production in Colombia.(AU)

Plasmid-Mediated Antibiotic Resistance and Class 1 Integron in Salmonella Heidelberg Isolated from Poultry Farms in Santander - Colombia

Salmonella enterica is a zoonotic pathogen transmitted to humans by the consumption of contaminated poultry products. Salmonella causes around 93 million cases of gastroenteritis and 155,000 deaths worldwide. A high number of multidrug-resistant Salmonella isolates has been found from different segments of poultry production, and it is often associated with horizontal antibiotic resistance gene transfer through mobile elements such as plasmids, integrons, and transposons. The emergence of antibiotic-resistant Salmonella has been associated with the misuse of antibiotics in animal production and it is classified as a highly important pathogen from public health due to its zoonotic character and its high dissemination ability. This study aimed to determine the antibiotic resistance associated with plasmids, and class 1 and 2 integrons from Salmonella Heidelberg isolates from healthy chickens in poultry farms from Santander, Colombia. 15 Salmonella Heidelberg isolates obtained from cloacal samples, were evaluated through endpoint PCR to determine the presence of plasmid-encoded resistance genes, and class 1 and 2 integrons. The bla CMY2 , strA and strB, sul1 and sul2, and tetB genes were found in all 15 Salmonella Heidelberg. The class 1 integron was present in 11 out of 15 isolates, harbored the gene cassette dfra7.The results indicate the presence of a high number of antibiotic resistance genes associated with plasmids and class 1 integrons in Salmonella Heidelberg strains isolated from poultry farms, resulting in a public health concern, both in humans and poultry production in Colombia.

Diversity and antimicrobial resistance of Salmonellaentericaserovars in surface river water and sediment / Diversidade e resistência antimicrobiana de sorovares de Salmonella enterica em águas superficiais de rios e sedimentos

This study aimedto evaluate the contamination by Salmonellasp. in the Capinzal River, to determinethe prevalent serovars, patterns of antimicrobial resistance, and the genetic relationships between the serovars identified. A total of 108 samples were collected from 2016 to 2018. The isolation of Salmonellaspp. was conducted accordingto InternationalOrganization for Standardization (ISO) standards. The antimicrobial resistance profile of the Salmonella isolates was evaluated, and isolates were selected for serotyping and verification of genetic similarity using the Pulsed-FieldGel Electrophoresis (PFGE) Technique. Of the 108 samples collected, 35 (32.4%) were positive for Salmonella; 17.2% of the isolates were from the rural area; and 88.6% were from the urban area. Salmonellawas isolated from all collect points along the river, with a higher incidence at the beginning of the urban area, indicating that contamination starts in the rural area and intensifies in the urban area of the city. A percentageof 35.1% of the Salmonellaisolates were resistant to at least two antibiotics, while 18.9% were considered multidrug-resistant (resistant to at least two antibiotics of different classes). Seven serovars were distinguished from the serotyped isolates, with a prevalence rate of 23.5% for S.Infantis, S.Orion, and S. Javiana; 11.8% forS.Senfterberg, and 5.9% forS. Montevideo, S.Heidelberg, and S.entericasubsp. enterica(O: 6.8). The variability in specific restriction sites generated by PFGE resulted in 10 pulsotypes, separating mainly different serotypes.

O objetivo deste estudo foi avaliar a contaminação por Salmonellasp. no rio Capinzal, para determinar os sorovares prevalentes, padrões de resistência antimicrobiana e as relações genéticas entre os sorovaresidentificados. Um total de 108 amostras foram coletadas de 2016 a 2018. O isolamento de Salmonellaspp. foi conduzido de acordo com os padrões da International Organization for Standardization (ISO). O perfil de resistência antimicrobiana dos isolados de Salmonellafoi avaliado, e os isolados foram selecionados para sorotipagem e verificação de similaridade genética por meio da Técnica de Eletroforese em Gel de Campo Pulsado (PFGE). Das 108 amostras coletadas, 35 (32,4%) foram positivaspara Salmonella; 17,2% dos isolados eram da área rural; e 88,6% da área urbana. Salmonellafoi isolada em todos os pontos de coleta ao longo do rio, com maior incidência no início da área urbana, indicando que a contaminação começa na área rural e se intensifica na área urbana da cidade. Um percentual de 35,1%dos isolados de Salmonellaforam resistentes a pelo menos dois antibióticos, enquanto 18,9% foram considerados multirresistentes (resistentes a pelo menos dois antibióticos de classes diferentes). Sete sorovares foram diferenciadosdos isolados sorotipados, com uma taxa de prevalência de 23,5% para S.Infantis,S.Orion e S.Javiana; 11,8% para S.Senfterberg e 5,9% para S.Montevidéu, S.Heidelberg e S.enterica subsp. enterica (O: 6,8). A variabilidade emlocais de restrição específicos gerados por PFGE resultou em 10 pulsotipos, separando principalmente diferentes sorotipos.

Diversity and antimicrobial resistance of Salmonellaentericaserovars in surface river water and sediment / Diversidade e resistência antimicrobiana de sorovares de Salmonella enterica em águas superficiais de rios e sedimentos

This study aimedto evaluate the contamination by Salmonellasp. in the Capinzal River, to determinethe prevalent serovars, patterns of antimicrobial resistance, and the genetic relationships between the serovars identified. A total of 108 samples were collected from 2016 to 2018. The isolation of Salmonellaspp. was conducted accordingto InternationalOrganization for Standardization (ISO) standards. The antimicrobial resistance profile of the Salmonella isolates was evaluated, and isolates were selected for serotyping and verification of genetic similarity using the Pulsed-FieldGel Electrophoresis (PFGE) Technique. Of the 108 samples collected, 35 (32.4%) were positive for Salmonella; 17.2% of the isolates were from the rural area; and 88.6% were from the urban area. Salmonellawas isolated from all collect points along the river, with a higher incidence at the beginning of the urban area, indicating that contamination starts in the rural area and intensifies in the urban area of the city. A percentageof 35.1% of the Salmonellaisolates were resistant to at least two antibiotics, while 18.9% were considered multidrug-resistant (resistant to at least two antibiotics of different classes). Seven serovars were distinguished from the serotyped isolates, with a prevalence rate of 23.5% for S.Infantis, S.Orion, and S. Javiana; 11.8% forS.Senfterberg, and 5.9% forS. Montevideo, S.Heidelberg, and S.entericasubsp. enterica(O: 6.8). The variability in specific restriction sites generated by PFGE resulted in 10 pulsotypes, separating mainly different serotypes.(AU)

O objetivo deste estudo foi avaliar a contaminação por Salmonellasp. no rio Capinzal, para determinar os sorovares prevalentes, padrões de resistência antimicrobiana e as relações genéticas entre os sorovaresidentificados. Um total de 108 amostras foram coletadas de 2016 a 2018. O isolamento de Salmonellaspp. foi conduzido de acordo com os padrões da International Organization for Standardization (ISO). O perfil de resistência antimicrobiana dos isolados de Salmonellafoi avaliado, e os isolados foram selecionados para sorotipagem e verificação de similaridade genética por meio da Técnica de Eletroforese em Gel de Campo Pulsado (PFGE). Das 108 amostras coletadas, 35 (32,4%) foram positivaspara Salmonella; 17,2% dos isolados eram da área rural; e 88,6% da área urbana. Salmonellafoi isolada em todos os pontos de coleta ao longo do rio, com maior incidência no início da área urbana, indicando que a contaminação começa na área rural e se intensifica na área urbana da cidade. Um percentual de 35,1%dos isolados de Salmonellaforam resistentes a pelo menos dois antibióticos, enquanto 18,9% foram considerados multirresistentes (resistentes a pelo menos dois antibióticos de classes diferentes). Sete sorovares foram diferenciadosdos isolados sorotipados, com uma taxa de prevalência de 23,5% para S.Infantis,S.Orion e S.Javiana; 11,8% para S.Senfterberg e 5,9% para S.Montevidéu, S.Heidelberg e S.enterica subsp. enterica (O: 6,8). A variabilidade emlocais de restrição específicos gerados por PFGE resultou em 10 pulsotipos, separando principalmente diferentes sorotipos.(AU)

Antimicrobial activity and chemical characterization of Laurel essential oil / Atividade antimicrobiana e caracterização química do óleo essencial de Louro / Actividad antimicrobiana y caracterización química del aceite esencial de Laurel

The factors that arouse interest in the study of essential oils as biocidal agents are numerous, such as the fact that they have antibacterial, antifungal, insecticidal, antioxidant, anti-inflammatory and larvicidal properties. The objective of this work was to evaluate the antimicrobial activity, in vitro, of the laurel (Laurus nobilis L) essential oil on the growth of pathogenic bacteria Salmonella enterica serovar Typhimurium ATCC 14028 and Staphylococcus aureus ATCC 25923, at different exposure times, as well as to perform the chemical characterization. Twenty compounds were identified and quantified, representing 96.57% of the total composition. The class of oxygenated monoterpenes represented the majority class of the essential oil, with 1,8-cineol (33.8%) as the substance found in greater quantity, followed by linalool (17.79%). The third constituent in greater quantity was sabinene (12.23%), belonging to the group of monoterpene hydrocarbons. Terpinyl acetate (9.41%) was also considered to be quantitatively representative. Laurel essential oil showed bacteriostatic activity against S. Typhimurium ATCC 14028 and S. aureus ATCC 25923.(AU)

Os fatores que despertam interesse no estudo dos óleos essenciais como agentes biocidas são inúmeros, como o fato de possuírem propriedades antibacteriana, antifúngica, inseticida, antioxidante, antiinflamatória e larvicida. O objetivo deste trabalho foi avaliar a atividade antimicrobiana, in vitro, do óleo essencial de louro (Laurus nobilis L) sobre o crescimento das bactérias patogênicas Salmonella enterica sorovar Typhimurium ATCC 14028 e Staphylococcus aureus ATCC 25923, em diferentes tempos de exposição, assim como realizar a caracterização química do óleo. Vinte compostos foram identificados e quantificados, representando 96,57% da composição total. A classe dos monoterpenos oxigenados representou a classe majoritária do óleo essencial, sendo o 1,8-cineol (33,8%) a substância encontrada em maior quantidade, seguido do linalol (17,79%). O terceiro constituinte em maior quantidade foi o sabineno (12,23%), pertencente ao grupo dos hidrocarbonetos monoterpênicos. O acetato de terpinila (9,41%) também foi considerado quantitativamente representativo. O óleo essencial de louro apresentou atividade bacteriostática contra S. Typhimurium ATCC 14028 e S. aureus ATCC 25923.(AU)

Los factores que despiertan interés en el estudio de los aceites esenciales como agentes biocidas son innumerables, como el hecho de que tienen propiedades antibacterianas, anti fúngicas, insecticidas, antioxidantes, antiinflamatorias y larvicidas. El objetivo de este trabajo ha sido evaluar la actividad antimicrobiana, in vitro, del aceite esencial de laurel (Laurus nobilis L) sobre el crecimiento de bacterias patógenas Salmonella enterica serovar Typhimurium ATCC 14028 y Staphylococcus aureus ATCC 25923, en diferentes momentos de exposición, así como realizar la caracterización química del aceite. Se identificaron y cuantificaron veinte compuestos, que representan el 96,57% de la composición total. La clase de mono terpenos oxigenados representó la clase principal de aceite esencial, siendo el 1,8-cineol (33,8%) la sustancia que se encuentra en mayor cantidad, seguida del linalol (17,79%). El tercer constituyente en mayor cantidad fue el sabineno (12,23%), perteneciente al grupo de los hidrocarburos monoterpénicos. El acetato de terpinilo (9,41%) también se consideró cuantitativamente representativo. El aceite esencial de laurel mostró actividad bacteriostática contra S. Typhimurium ATCC 14028 y S. aureus ATCC 25923.(AU)

Symbiotic drink based on Brazil nuts (Bertholletia excelsa H. B. K): production, characterization, probiotic viability and sensory acceptance / Bebida simbiótica à base de castanha-do-Brasil (Bertholletia excelsa H. B. K): produção, caracterização, viabilidade probiótica e aceitação sensorial

Brazil nut is a typical oilseed from the Amazon region, of great economic and cultural importance. Due to its high nutritional value, it is interesting to be used as a matrix for new products development. Thus, the present study aimed to develop and characterize a drink based on Brazil nuts fermented by Lactobacillus casei and added with inulin. After formulated, the drink was pasteurized, fermented, maturated, and stored under refrigeration for 28 days. Physical, chemical, microbiological, and sensory analyzes were performed before and after fermentation, at 7-day intervals. There was maintenance in the content of lipids and proteins and reduction in the concentrations of fibers, ash, total carbohydrates, and moisture. The fermented drink pH decreased significantly during storage, while the acidity and the soluble solids content increased from the 14th day. Regarding the color, there were significant changes in all parameters analyzed. In the fermented drink, the development of molds and yeasts, total and thermotolerant coliforms, and Salmonella were not verified, while the control drink deteriorated after 7 days. The probiotic L. casei had count ranging from 9.48 to 8.59 log CFU mL-1 during the 28 days. When exposed to in vitro gastrointestinal conditions, L. casei significantly reduced its concentration. However, it reached the enteric phase with counts greater than 6.00 log UFC mL-1. Sensorially, the drink reached intermediate scores (between 5.41 and 6.02). Therefore, the use of Brazil nuts proved to be viable for the development of a product free of animal-origin components, symbiotic, and of outstanding nutritional quality.(AU)

A castanha-do-Brasil é uma oleaginosa típica da região Amazônica, de grande importância econômica e cultural. Devido ao seu alto valor nutritivo, torna-se interessante a sua utilização como matriz para o desenvolvimento de novos produtos. Assim, o presente estudo teve como objetivo o desenvolvimento e caracterização de uma bebida à base de castanha-do-Brasil fermentada por Lactobacillus casei e adicionada de inulina. Após formulada, a bebida foi pasteurizada, fermentada, maturada e armazenada sob refrigeração por 28 dias. Foram realizadas análises físicas, químicas, microbiológicas e sensoriais antes e após a fermentação, em intervalos de sete dias. Observou-se uma manutenção no teor de lipídeos e proteínas e redução nas concentrações de fibras, cinzas, carboidratos totais e umidade. O pH da bebida fermentada reduziu significativamente ao longo do armazenamento, enquanto a acidez e o teor de sólidos solúveis aumentaram a partir do 14° dia. Em relação à cor, houve alterações significativas em todos os parâmetros analisados. Na bebida fermentada, não foi verificado o desenvolvimento de bolores e leveduras, coliformes totais e termotolerantes e Salmonella, enquanto a bebida controle se deteriorou após sete dias. O probiótico L. casei apresentou contagens variando de 9.48 a 8.59 log CFU mL-1 durante os 28 dias de armazenamento. Quando exposto às condições gastrointestinais in vitro, o L. casei diminui a sua concentração significativamente. Contudo, atingiu a fase entérica com contagens superiores a 6.00 log UFC mL-1. Sensorialmente, a bebida alcançou scores intermediários (entre 5.41 e 6.02). Dessa forma, a utilização da castanha-do-Brasil se mostrou viável para o desenvolvimento de um produto isento de componentes de origem animal, simbiótico e de destacável qualidade nutricional.(AU)