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.

Effect of Ammonia Gas in Poultry Litter Contaminated with Salmonella Heidelberg

Salmonella Heidelberg is an emerging pathogen in Brazilian poultry production. The traditional methods (quicklime, windrowing and tarpaulin-on-surface) used for disinfecting reused poultry litter between flocks does not guarantee its elimination, thus allowing the transmission of this agent from one flock to another. The new tarpaulinon-surface method with controlled injection of ammonia gas has proven to be effective in its control, however, it is still unknown what dose of ammonia gas is needed to eliminate Salmonella Heidelberg in reused poultry litter. The objective of this study was to evaluate the effect of ammonia gas at different concentrations in sterile poultry litter artificially contaminated with Salmonella Heidelberg. Then, ammonia gas was injected in concentrations of 0.25%, 0.5%, and 1%, and 48 hours later, a sample was collected from each repetition in an entirely randomized design, and bacterial isolation was performed. All treatments, including positive and negative controls, were tested in quadruplicate and the parameters temperature, humidity, pH and water activity were evaluated. In the 0.5% and 1% treated samples the pathogen was not isolated, while in the 0.25% concentration one of the four samples tested was positive. The study reveals that ammonia gas is efficient in killing Salmonella Heidelberg in poultry litter at concentrations of 0.5 % or more within a 48-hour period and that the litter treated with ammonia gas increases its pH and water activity.(AU)

Performance, Gut Integrity, Enterobacteria Content in Ceca of Broiler Fed Different Eubiotic Additives

An experiment was carried out to study the effect of different eubiotics on productive characteristics, intestinal integrity, as well as the content of enterobacteria in the cecum of broiler chickens. A completely randomized design with five treatments and 8 replicates of 25 birds each was used. In total 1000 mixed broiler chickens from Ross308 strain, one day old were obtained from a commercial hatchery. The birds were housed on concrete floors in a conventional house. A sorghum+soybean meal control diet was used, to which the additives under study were added. The treatments were distributed as follows: T1 = Control diet without antibiotic or eubiotic; T2 = T1 + bacteriophages; T3 = T1 + antibiotic; T4 = T1 + probiotic; T5 = T1 + symbiotic. The results obtained at 49 days of age for weight gain and feed conversion rate improved (p<0.05) with the addition of the antibiotic and eubiotics. A lower (p<0.05) intestinal density was observed with the probiotic. The height, width, and area of villi in duodenum was higher (p<0.05) when antibiotic and eubiotics were included. In the histological score, in duodenum, the antibiotic and eubiotics resulted with a higher score (p<0.05), associated to a physiological and controlled inflammation response that allowed improving productivity. Finally, the relative expression of enterobacteria, such as Lactobacillus salivarius, allowed associating positive changes in the microbiome and better productive parameters when including the symbiotic, with comparable results to the antibiotic when including the eubiotics.(AU)

Ganoderma Lucidum Extract Modulates Cecal Microbiota Community in Broilers under Dextran Sulfate Sodium Challenge During the Starter Phase

In this study, we investigated the effects of Ganoderma lucidum extract (GLE) supplementation on the cecal microbiota of broilers challenged with dextran sulfate sodium (DSS) during the starter phase. A total of 32 one-day-old, unsexed broiler chicks were randomly divided into four dietary treatments with eight birds per treatment and reared individually for 14 days (n = 8). The diet treatments were: non-DSS challenge, DSS challenge only, DSS challenge plus 0.5 mL/L GLE, and DSS challenge plus 1 mL/L GLE. The results showed that DSS challenge plus 0.5 mL GLE alleviated inflammatory gene expression in the duodenum of broilers (p≤0.01). The alpha diversity of bacterial species in the cecal digesta increased in the group treated with DSS plus 1 mL/L GLE compared with the DSS challenge-only group (p≤0.01). Principal component analysis and principal coordinate analysis indicated distinct clusters between groups treated with DSS-only and DSS plus GLE (0.5 and 1 mL/L). The abundance of the genera Ruminiclostridium 9, Enterococcus, and Sellimonas increased in the group treated with DSS plus GLE (0.5 and 1 mL/L) compared with the other groups (p≤0.01). Comparative microbial function analysis demonstrated that the immune system was promoted in the group treated with DSS plus GLE (0.5 and 1 mL/L) compared to the DSS challenge-only group (p≤0.001). These results demonstrated that GLE supplementation can modulate the cecal microbial community of broilers under DSS challenge during the starter phase.(AU)

Comparative Study on the Predominance of Lactobacillus spp. and Escherichia Coli in Healthy vs Colibacillosis Diseased Broilers

This study aims to identify relative proportions of beneficial and pathogenic bacteria in the gut of broilers and risk factors that may be contributing to the development of colibacillosis disease in broiler farms of District Kasur, Punjab, Pakistan. For this, 10 healthy and 10 colibacillosis affected broiler farms were surveyed for ileum and blood sample collection along with data regarding farm management, antibiotic use and hygiene practices. Lactobacillus and Escherichia coli number was estimated using Miles and Misra method and colibacillosis was confirmed by Congo red dye assay. Lactobacillus and E. coli were identified biochemically. For risk factors analysis chi-square analysis was performed to find any significant association between the health status of the farm and risk factors. Results showed during disease and healthy conditions Lactobacillus and Escherichia coli counts differ significantly (p<0.05). E. coli counts (106-108 to 107-109) increased (p<0.05) about three folds and Lactobacillus counts decrease (106-108 to 105-107) about four folds in disease conditions. Risk factor analysis showed colibacillosis disease was significantly associated (p<0.05) with non-vaccinated flocks, natural ventilation systems, rodent presence and the lack of outfit disinfection or change by workers when moving between different houses. It is concluded that E. coli and Lactobacillus work antagonistically to each other. However, further research is necessary to determine the exact mechanisms by which E. coli and Lactobacillus influence the development of colibacillosis. While Lactobacillus as probiotic may help with prevention, good hygiene and management practices are still crucial in preventing the spread of disease.(AU)

Characterization of coccidiosis and evaluation of suggestive cases of subclinical necrotic enteritis in broilers / Caracterização de coccidiose e avaliação de casos sugestivos de enterite necrótica subclínica em frangos de corte

This study performed the characterization of coccidiosis in broilers and evaluated the occurrence of suggestive cases of necrotic enteritis (NE), seeking if there is an association between the diseases in Brazilian flocks. Two hundred and fifty-six birds from 32 flocks were evaluated. Macroscopic and histopathological lesions were graduated for coccidiosis and NE. Intestinal content was investigated by polymerase chain reaction (PCR) for seven species of Eimeria and by selective anaerobic culture for Clostridium perfringens and identification of the NetB gene. Flocks positive for coccidiosis represented 93.8%. Macroscopic lesions of coccidiosis were Grade 1 for E. acervulina (27%); E. tenella (9.7%) and E. maxima (8.9%). Histopathological evaluation showed Grade 1 in duodenum (38.2%); jejunum (21.4%); cecum (9.3%) and ileum (5%). PCR demonstrated positivity for E. tenella (21.9%), E. maxima (18.8%), and E. acervulina (3.1%). Suggestive macroscopic lesions of necrotic enteritis ranged from Grade 1 (16%), 2 (23%) and 3 (10,9%). Histopathology indicated the absence of necrosis, showing only hemorrhage in the mucosa and submucosa, with the presence of Eimeria spp. Clostridium perfringens type A netB+ was not isolated, demonstrating that macroscopic lesions found mostly in the jejunum did not characterize NE, based on histopathology and negativity of the NetB gene. The study suggests that, due to the high occurrence of coccidiosis, many macroscopic findings suggestive of NE are, in fact, attributed to atypical lesions caused by the reproduction of Eimeria spp.

Este estudo realizou a caracterização de coccidiose em frangos de corte e avaliou a ocorrência de casos sugestivos de enterite necrótica (EN), buscando se há alguma associação entre estas duas enfermidades em lotes de frango de corte no Brasil. Foram avaliadas 256 aves de 32 lotes. Lesões macroscópicas e histopatológicas foram graduadas para coccidiose e EN. O conteúdo intestinal foi investigado por reação em cadeia da polimerase (PCR) para sete espécies de Eimeria e por cultura anaeróbia seletiva para Clostridium perfringens e identificação do gene NetB. Os lotes positivos para coccidiose representaram 93,8%. Lesões macroscópicas de coccidiose foram de Grau 1 para E. acervulina (27%); E. tenella (9,7%) e E. maxima (8,9%). A avaliação histopatológica mostrou Grau 1 no duodeno (38,2%); jejuno (21,4%); ceco (9,3%) e íleo (5%). A PCR demonstrou positividade para E. tenella (21,9%), E. maxima (18,8%) e E. acervulina (3,1%). Lesões macroscópicas sugestivas de enterite necrótica variaram de grau 1 (16%), 2 (23%) e 3 (10,9%). A histopatologia indicou ausência de necrose, apresentando apenas hemorragia em mucosa e submucosa, com presença de Eimeria spp. Clostridium perfringens tipo A netB + não foi isolado, demonstrando que lesões macroscópicas encontradas principalmente no jejuno não caracterizaram NE, com base na histopatologia e negatividade do gene NetB. O estudo sugere que, em virtude da alta ocorrência de coccidiose nos lotes, muitos achados macroscópicos sugestivos de EN são, na verdade, atribuídos a lesões atípicas provocadas pela reprodução de Eimeria spp.

Genetic variability of Mycoplasma synoviae detected in commercial layers in southeastern Brazil / Variabilidade genética de Mycoplasma synoviae detectado em poedeiras comerciais do sudeste do Brasil

Este estudo teve como objetivo caracterizar genótipos de M. synoviae circulantes em poedeiras comerciais, na Região Sudeste do Brasil. Para estabelecer a relação evolutiva entre as cepas de MS, foi analisada a sequência genética do gene vlhA de 11 cepas de MS identificadas no Rio de Janeiro e em São Paulo, de 10 outras cepas de MS identificadas no Brasil e cujas sequências foram depositadas no banco de dados GenBank, e da cepa vacinal MSH. O método da máxima verossimilhança e o modelo de Kimura com dois parâmetros foram utilizados para comparar as cepas. As sequências obtidas foram depositadas no Genbank, sob os números de acesso OP279775 a OP279785. Foi possível verificar a presença de diferentes cepas circulantes no Brasil, com alta similaridade entre as cepas do Rio de Janeiro pela análise do gene vlhA. As duas cepas paulistas detectadas no presente estudo possuem o baixo percentual (68%) de similaridade, demonstrando a variabilidade das cepas dessa localidade.

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)

Effects of Medium Chain Fatty Acid Supplementation in Broiler Diet on Microbiological Quality of Litter

In our experiment, coconut and palm oil supplementation was added to the diet of broiler chickens to prove the hypothesis that medium chain fatty acids (MCFAs) can reduce the occurrence of potential pathogens (Clostridium perfingens, Coliform) in gut microflore and therefore improve bird welfare. Cobb 500 cockerels were divided in five groups. Control birds were fed commercial broiler diet, while birds in the four treatment group diets were supplemented with coconut, palm oil, or a combination of the two, respectively. As a positive control, sunflower oil supplementation was included in the diet of the fifth group. During the 28 days of the study (from 14 to 42 days of life), 5 samplings were scheduled, when excreta samples were taken from the litter to analyse total microbial count and the number of Clostridia, Coliforms, and Salmonella. According to the results of microbiological analysis, coconut oil supplementation led to the continuous decline of Clostridium perfringens numbers until they vanished by the end of the experimental period. A similar but faster decline was found as a result of palm oil and the combined (palm oil+coconut oil) treatment. However, the number of Coliforms and total microbial count changed only slightly by the end of the study. No Salmonella spp. was present in the samples throughout the experiment. Altogether, coconut and palm oil supplementation has a beneficial effect on the microbiological composition of poultry litter.(AU)

The influence of clove leave extract (Syzygium aromaticum) on growth performance and bacterial population of broiler chickens raised in stressful conditions of high stocking density

Broilers maintenance at high stocking density impacts uncomfortable conditions; this increases the potential for stress, reduces feed consumption, and reduces production performance and health in broiler chickens. This study aims to evaluate the effect of clove leaf extract on growth performance and microbial population in the digestive tract of broiler chickens reared at high density. From the eighth day, five treatment groups, T0 (negative control with a typical density of 10 birds/m2 ), T1 (positive control with a high density of 16 birds/m2 ), T2, T3, and T4 with a high density of 16 birds/m2, were randomly assigned from 444 broilers (bodyweight of 129.78±0.75 g) with six repetitions each on the eighth day. The feed includes clove leave extract as much as 0.5, 0.75, and 1 ml/kg for T2, T3, and T4, respectively. The weight gain, feed intake, and feed conversion ratio were tracked weekly. One chicken from each replicate was taken randomly, slaughtered, and feathered on day 35. The digesta was taken from the ileum and cecum to measure the intestinal bacterial population and put into a sterile container. Digesta from the duodenum, jejunum, ileum, and cecum were also collected to determine pH levels. MRS agar was used to determine the amount of LAB in the digesta. The findings revealed that giving clove leaf extract of feed to broiler chickens kept at high stocking density could higher body weight gain (P < 0.05), higher feed consumption (P < 0.05), decreased the pH of the ileum and cecum (P < 0.05), increased lactic acid bacteria and decreased coliform bacteria in the ileum and cecum (P < 0.05).

Effects of Medium Chain Fatty Acid Supplementation in Broiler Diet on Microbiological Quality of Litter

In our experiment, coconut and palm oil supplementation was added to the diet of broiler chickens to prove the hypothesis that medium chain fatty acids (MCFAs) can reduce the occurrence of potential pathogens (Clostridium perfingens, Coliform) in gut microflore and therefore improve bird welfare. Cobb 500 cockerels were divided in five groups. Control birds were fed commercial broiler diet, while birds in the four treatment group diets were supplemented with coconut, palm oil, or a combination of the two, respectively. As a positive control, sunflower oil supplementation was included in the diet of the fifth group. During the 28 days of the study (from 14 to 42 days of life), 5 samplings were scheduled, when excreta samples were taken from the litter to analyse total microbial count and the number of Clostridia, Coliforms, and Salmonella. According to the results of microbiological analysis, coconut oil supplementation led to the continuous decline of Clostridium perfringens numbers until they vanished by the end of the experimental period. A similar but faster decline was found as a result of palm oil and the combined (palm oil+coconut oil) treatment. However, the number of Coliforms and total microbial count changed only slightly by the end of the study. No Salmonella spp. was present in the samples throughout the experiment. Altogether, coconut and palm oil supplementation has a beneficial effect on the microbiological composition of poultry litter.

Surto de botulismo tipo C em aves de subsistência em Santa Catarina, Brasil / Outbreak of type C botulism in backyard poultry in Santa Catarina, Brazil

Background: Botulism is a disease caused by the ingestion of neurotoxin produced by Clostridium botulinum, characterizedby flaccid paralysis, which can lead to high mortality. They have seven types of neurotoxins (A, B, C, D, E, F, and G) and,in birds, most cases are attributed to type C. They are considered sources of botulinum toxins where the decomposition oforganic matter occurs, like stagnant water and rotting food. The main feature of the disease in birds is ascending symmetricflaccid paralysis. The present study aims to describe an outbreak of type C botulism in backyard poultry in the state ofSanta Catarina, Southern Brazil.Case: A visit was made to the property with 160 backyard poultry with a history of high mortality in the municipality ofAgrolândia, Santa Catarina. Clinical signs were characterized by paralysis of the pelvic limbs, neck and pendular wings,which progressed to death within 48 h. There was a mortality rate of 37.5% (60/160) between March and May 2019. Thesebirds were kept in an overcrowded environment, with different species (chickens, ducks, teals, and turkeys) fed irregularly.The water supplied was provided from kitchen exhaust, accumulating in puddles on the floor that contained organic matterresidues such as animal feces, food waste and bone fragments. The disposal of the carcasses of birds that died was in thesame enclosure, buried superficially, facilitating the access of other birds to dig them up and consume them. Necropsywas performed on 2 chickens and one duck, no macroscopic or histopathological lesions were observed. Blood, liver, andgastrointestinal content samples were sent for research and identification of botulinum toxin through the serum neutralization test in mice. The presence of type C botulinum toxin was confirmed in the liver chicken of one sampled animals.Discussion: The identification of type C botulism toxin enabled the characterization of the outbreak, which is...

Surto de botulismo tipo C em aves de subsistência em Santa Catarina, Brasil / Outbreak of type C botulism in backyard poultry in Santa Catarina, Brazil

Background: Botulism is a disease caused by the ingestion of neurotoxin produced by Clostridium botulinum, characterizedby flaccid paralysis, which can lead to high mortality. They have seven types of neurotoxins (A, B, C, D, E, F, and G) and,in birds, most cases are attributed to type C. They are considered sources of botulinum toxins where the decomposition oforganic matter occurs, like stagnant water and rotting food. The main feature of the disease in birds is ascending symmetricflaccid paralysis. The present study aims to describe an outbreak of type C botulism in backyard poultry in the state ofSanta Catarina, Southern Brazil.Case: A visit was made to the property with 160 backyard poultry with a history of high mortality in the municipality ofAgrolândia, Santa Catarina. Clinical signs were characterized by paralysis of the pelvic limbs, neck and pendular wings,which progressed to death within 48 h. There was a mortality rate of 37.5% (60/160) between March and May 2019. Thesebirds were kept in an overcrowded environment, with different species (chickens, ducks, teals, and turkeys) fed irregularly.The water supplied was provided from kitchen exhaust, accumulating in puddles on the floor that contained organic matterresidues such as animal feces, food waste and bone fragments. The disposal of the carcasses of birds that died was in thesame enclosure, buried superficially, facilitating the access of other birds to dig them up and consume them. Necropsywas performed on 2 chickens and one duck, no macroscopic or histopathological lesions were observed. Blood, liver, andgastrointestinal content samples were sent for research and identification of botulinum toxin through the serum neutralization test in mice. The presence of type C botulinum toxin was confirmed in the liver chicken of one sampled animals.Discussion: The identification of type C botulism toxin enabled the characterization of the outbreak, which is...(AU)

Expression of TGF-ß/Smads in Cecum and Spleen of Chicken Infected with E. Tenella

Chicken coccidiosis is a common and severe parasitic disease caused by infection from Eimeria spp., which affects the integrity of the intestinal mucosa. TGF-ß has been shown to play an important role in the healing of intestinal mucosas, immunity, and the maintenance of bowel mucosa integrity. Very little is known about the presence of the components of TGF-ß/Smads signaling pathway of chicken at different times following coccidian infection. In the present study, we measured the levels of TGF-ß2, 3, 4, receptor TßRI, II, down-stream Smad 2, 3, 7 in cecum and spleen of chicken at different times after inoculation with Eimeria tenella using quantitative real-time PCR. The results showed that the TGF-ß/Smads signaling pathway was not activated in cecum in the early stage of infection. However, on the 8th day, the expression of TGF-ß2, 4, down-stream protein Smad 2, 7 were significant up-regulated in the spleen, which indicated that the TGF-ß/Smads signaling was changed in the E. tenella infection and was differentially expressed in various tissues in the early stages of infection.(AU)

Inhibitory effect of different probiotic formulations on Salmonella Heidelberg, avian pathogenic Escherichia coli and Campylobacter jejuni

A wide variety of microorganism species have been used as probiotics to improve the intestinal microbial balance, control and prevent the colonization of pathogenic bacteria and promote growth in animal production. Based on the various beneficial factors of probiotic agents, this study aimed to evaluate the effectiveness of three different formulations of probiotics strains through the analysis of antagonistic capacity against common pathogens from chickens gastrointestinal tract. Three formulations composed by Bacillus, Lactobacillus, Saccharomyces, Enterococcus, Bifidobacterium and Pediococcus were tested in vitro by the method of probiotic culture spot in plates seeded with inocula of Escherichia coli, Salmonella Heidelberg and Campylobacter jejuni. The inhibition halos were measured and classified according to the degree of pathogenic bacteria inhibition. The formulation containing an association among Bacillus, Lactobacillus and Saccharomyces presented better results when compared to the other formulations.

Direct Detection and Quantification of Bacterial Pathogens from Broiler Cecal Samples in the Slaughter Line by Real-Time PCR

Chicken meat is an important source of foodborne pathogens, including Salmonella, Campylobacter, and Clostridium perfringens. These bacteria can occur in the intestinal microbiota of broilers and contaminate chicken carcasses in industrial meat processing. This study aimed to develop and evaluate a procedure based on real-time PCRs for the direct detection and quantification of these three bacteria in broilers' ceca collected in poultry slaughter houses and demonstrate the occurrence of these important foodborne pathogens in Brazilian poultry production flocks. Cecal contents were collected from 45 different broiler flocks in three different slaughterhouses in the state of Paraná, Brazil, totaling 45 samples (in pools of 10 different ceca/chickens per broiler flock). Then, these samples were tested for the detection and quantification of Salmonella, Campylobacter, and Clostridium perfringens by real-time PCRs. The results demonstrated the occurrence of three (6.7%) positive pools for Salmonella, 20 (44.4%) for Campylobacter, and 32 (71.1%) for C. perfringens. Mean bacterial concentrations in the positive samples were 4.3log10 cells/g for Salmonella, 6.4 log10 cells/g for Campylobacter, and 5.5 log10 cells/g for C. perfringens. In conclusion, Salmonella, Campylobacter, and C. perfringens could be detected and quantified directly from the broilers cecal contents collected in the slaughter line. This procedure will be certainly useful to more quickly detect these foodborne pathogens and prevent their occurrence in chicken meat and other poultry food products.(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

Detection of Mycoplasma Synoviae and Other Pathogens in Laying Hens with Respiratory Signs in the Rearing and Production Phases

The present study aimed to investigate the occurrence of Mycoplasma synoviae (MS), M. gallisepticum (MG), Ornitobacterium rhinotracheale (OR), Avibacterium paragallinarum (AP), Pasteurella multocida (PM) and Infectious Bronchitis Virus (IBV) in laying hens with respiratory clinical signs in two phases of production. 140 tracheal swabs and 140 blood samples were collected from laying hens in the rearing and production phases, the chickens belonged to six farms (A-F) located in the state of São Paulo, Brazil. The samples were analyzed by PCR for MG, MS, OR, AP, PM and IBV and by ELISA for MG and MS. The highest frequencies observed by PCR were for MS at farms B and C with 95 and 100% positivity, followed by MG at farms D and E with 35% and 65%, IBV with 35% at farm F and ORT with 15% at farm A. All flocks were positive for MG and MS in serology. Although MG and IBV have been detected, this can be explained by the vaccination protocols, since live attenuated vaccines are widely used for immunization against these pathogens. It was also possible to detect OR and AP thorugh PCR in some flocks. The occurrence of several etiological agents that cause respiratory diseases in laying hens was confirmed by PCR and serology, with MS being the most prevalent and being present in all farms studied.(AU)

Isolation, characterization, and resistance profile of Salmonella spp. from chicken cuts / Isolamento, caracterização e perfil de resistência de Salmonella spp. provenientes de cortes de frango

The present study aimed at isolating and characterizing Salmonella spp. from chicken cuts marketed in Francisco Beltrão, PR, and verify the resistance profile of the isolates against antimicrobials used in human therapy. Samples of chicken cuts (n=40) were purchased from supermarkets and submitted to microbiological analysis for the detection of Salmonella spp. The suspected colonies underwent biochemical testing for the identification of enterobacteria. Four colonies were selected from each sample positive for Salmonella spp., totaling 28 isolates that were tested for antimicrobial sensitivity. Colonies that showed resistance to ceftriaxone were subjected to extended-spectrum beta-lactamases (ESBL). Among the analyzed chicken samples, seven (17.5%) showed biochemical behavior characteristic of Salmonella spp. Among the 28 isolates, seventeen different resistance profiles were found, of which 46.42% (n=13) had a multi-resistance profile, and 21.4% (n=6) of the isolates had a phenotype for ESBL production. The strains of Salmonella spp. isolated from chicken cuts found in this study showed a high level of resistance to antimicrobials of different classes and of last generations, these data serve as a warning, as they put the human treatment of salmonellosis at risk.(AU)

A pesquisa objetivou isolar e caracterizar Salmonella spp., a partir de cortes de frangos comercializados na cidade de Francisco Beltrão - PR, bem como verificar o perfil de resistência dos isolados em relação aos antimicrobianos utilizados na terapêutica humana. Amostras de cortes de frango (n=40) foram adquiridas em supermercados e submetidas à análise microbiológica para detecção de Salmonella spp. As colônias suspeitas foram submetidas a provas bioquímicas para identificação de enterobactérias. Quatro colônias foram selecionadas de cada amostra positiva para Salmonella spp., totalizando 28 isolados, que foram testadas quanto à sensibilidade a antimicrobianos. As colônias que apresentaram resistência à ceftriaxona foram submetidas à pesquisa de betalactamases de espectro estendido (ESBL). Das amostras de frango analisadas, sete (17,5%) apresentaram comportamento bioquímico característico de Salmonella spp. Entre os 28 isolados, foram encontrados 17 perfis diferentes de resistência, tendo 46,42% (n=13) apresentado perfil de multirresistência e 21,4% (n=6) apresentado fenótipo para produção de ESBL. As cepas de Salmonella spp. isoladas de cortes de frango, encontradas neste estudo, apresentaram alto índice de resistência a antimicrobianos de diferentes classes e de últimas gerações. Esses dados servem de alerta, uma vez que coloca em risco o tratamento da salmonelose humana.(AU)