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Despite attempts to control avian mycoplasmosis through management, vaccination, and surveillance, Mycoplasma gallisepticum continues to cause significant morbidity, mortality, and economic losses in poultry production. Live attenuated vaccines are commonly used in the poultry industry to control avian mycoplasmosis; unfortunately, some vaccines may revert to virulence and vaccine strains are generally difficult to distinguish from natural field isolates. In order to identify genome differences among vaccine revertants, vaccine strains, and field isolates, whole-genome sequencing of the M. gallisepticum vaccine strain ts-11 and several “ts-11-like” strains isolated from commercial flocks was performed using Illumina and 454 pyrosequencing and the sequenced genomes compared to the M. gallisepticum R low reference genome. The collective contigs for each strain were annotated using the fully annotated Mycoplasma reference genome. The analysis revealed genetic differences among vlhA alleles, as well as among genes annotated as coding for a cell wall surface anchor protein ( mg0377 ) and a hypothetical protein gene, mg0359 , unique to M. gallisepticum ts-11 vaccine strain. PCR protocols were designed to target 5 sequences unique to the M. gallisepticum ts-11 strain: vlhA 3.04a, vlhA 3.04b, vlhA 3.05, mg0377 , and mg0359 . All ts-11 isolates were positive for the five gene alleles tested by PCR; however, 5 to 36% of field isolates were also positive for at least one of the alleles tested. A combination of PCR tests for vlhA 3.04a, vlhA 3.05, and mg0359 was able to distinguish the M. gallisepticum ts-11 vaccine strain from field isolates. This method will further supplement current approaches to quickly distinguish M. gallisepticum vaccine strains from field isolates.

The role of invasive Salmonella in contamination of ground poultry is poorly defined. Salmonella harborage sites were determined in experimentally infected chickens and turkeys. Bioluminescent-tagged Salmonella were used to follow their spread in bone, meat, and skin following infection. Immunohistochemistry and culture were used to localize Salmonella. Chicken neck skin was positive for S. Heidelberg and S. Typhimurium throughout the experimental period, and the bacteria were localized on the stratum corneum of the epidermis and feather follicles. S. Heidelberg and S. Typhimurium were intermittently detected in drumstick muscle of chickens, with Salmonella primarily localized in connective tissues and lymphatics. Twenty percent of the drumstick muscles were culture positive for Salmonella in chickens at 42 days of age. Blood and tibiotarsus bone were culture positive for S. Heidelberg and S. Typhimurium during the first 2 weeks of infection. Salmonella levels in neck skin and muscle were

O objetivo deste estudo foi avaliar a resposta imunológica de frangos de corte frente a uma vacina vetorial HVT-IBV com posterior desafio com a cepa Moulthrop G603 aos 21 ou 28 dias de vida. Foram realizadas análises macroscópicas e histológicas da bolsa cloacal e quantificação de células imunológicas no baço (linfócitos B, T CD4+ e CD8+), bolsa cloacal (linfócitos T CD3+) e no sangue (linfócitos B, T CD4 de mucosa e T CD8 ativados). Aos 35 dias de idade, aves desafiadas aos 21 e 28 dias de idade, submetidas ou não a vacina vetorial, apresentaram maior contagem (P

SUMMARY Salmonellosis is one of the most common worldwide zoonosis, and the Enteritidis and Typhimurium serotypes are most prevalent. In this study, we evaluated the effects of a diet supplemented with mannanoligosaccharides (MOS) on CD3+ ,C D4+ ,C D8+, and goblet cell counts in the intestines of broilers vaccinated and challenged with Salmonella Enteritidis. The evaluation was based on randomized experiments including 45 chickens divided into 3 experimental groups. Group 1 received a nonsupplemented control diet. Group 2 received a diet supplemented with 1 kg MOS/t from d 1 to 21 and 0.5 kg MOS/t from d 22 to 56. Group 3 received a diet supplemented with 2 kg MOS/t from d 1 to 21 and 1 kg MOS/t from d 22 to 56. Chickens fed the lower level of MOS demonstrated reduced fecal shedding of Salmonella 14 d after being challenged (P < 0.05). Both groups of MOS-supplemented chickens showed an increase in CD4+ and CD8+ cell counts in the ileal and cecal mucosa after being challenged with Salmonella Enteritidis (SE). Supplementation with MOS increased the specific T lymphocyte infiltration in the intestinal mucosa of chickens vaccinated and challenged with Salmonella Enteritidis.

SUMMARY The role of the gastrointestinal tract in the recognition and elimination of pathogenic microorganisms through inflammation is critical for the establishment of health. Sanguinarine is a quaternary ammonium salt that belongs to a group of benzophenanthridine and protopin alkaloids. The present study was conducted to evaluate the effects of using sanguinarine in the drinking water of broilers on reducing the Salmonella Enteritidis count and to assess changes in intestinal morphology and the expression of immune cells in the peripheral blood and intestinal mucosa of broilers challenged with Salmonella Enteritidis. The treatment of Salmonella Enteritidis-challenged broilers with sanguinarine via drinking water reduced Salmonella Enteritidis isolation in the cecum and in the crop at 7 d postinoculation when compared with the control group. Broilers treated with sanguinarine via drinking water presented a significantly lower expression of goblet and CD3 + cells in the duodenum and jejunum and higher expression of cells positive for the markers cD4, cD8α, cD8α bright , cD8α dim , cD8β, tcr Vβ1, and cD28 in the blood compared with the nontreated birds. We also evaluated MHc i, MHc ii and tcr αβ Vβ2, but no differences were observed between treatments.

SUMMARY Different periods of hatching, called hatch windows, occur inside the same hatchery and birds’ development could be affected by this. This study evaluated morphology of the intestinal mucosa and gizzard, and the presence of CD3+ cells in thymus, spleen, and intestinal mucosa in broiler chicks hatched from different egg weights and in different hatch windows. Eggs from breeders at 38 wk of age were evaluated in a factorial 3 × 2 experimental design, with 3 hatch windows (hatching before 472 h of incubation, between 472 and 488 h of incubation, and between 488 and 504 h of incubation) and 2 egg weights (lightweight and heavyweight). Prematurely hatched chicks remaining inside the hatchery for longer periods presented improved koilin membrane development in the gizzard, and smaller presence of CD3+ cells in thymus, spleen, and ileum compared with those birds that hatched close to the standardized time for hatchery opening.

Para avaliar o efeito do probiótico sobre a resposta imunológica de frangos de corte desafiados com Salmonella Minnesota (SM), 60 frangos foram divididos em três grupos: CN- (controle negativo) aves que não foram inoculadas com SM, CP- (controle positivo) aves inoculadas com SM e Probiótico- aves suplementadas na ração com probiótico composto de Lactobacillus acidophilus, L. plantarium, L. rhamnosus, L. bulgaricus, Enterococcus faecium, Streptococcus thermophilus e Bifidobacterium bifidum e desafiadas com SM. Aos 14 dias foi realizada a inoculação com SM e aos 7 e 35 dias foram quantificadas células caliciformes, CD4+ e CD8+ na mucosa intestinal do íleo e ceco. Aves suplementadas com probióticos aos 7 dias de idade apresentaram aumento significativo (P≤0,05) de células caliciformes e CD4+ no íleo e de células CD8+ no ceco. Aos 35 dias houve aumento significativo (P≤0,05) das células CD8+ nas aves inoculadas do CN e Probiótico. A utilização de probióticos proporcionou redução significativa (P≤0,05) da contagem de Salmonella sp.

The use of Bacillus subtilis (BS) as a probiotic in bird feed was studied through the evaluation of its effect on th e infiltration of immune cells in the ileum and cecum mucosa of chickens challenged with Salmonella Minnesota (SM). The birds were divided into three treatment groups; Negative control, containing unchallenged birds; Positive control, with SM challenged birds; and Probiotic, with SM challenged birds fed with a diet containing BS (DSM 17299 2.13 x 10 cfu/g). The birds fed BS showed increased goblet and CD4+ 6 cell counts in the ileum and cecum before being challenged with SM in comparison to the birds not fed BS. After the SM challenge, the birds fed BS showed a reduction in the Salmonella counts at 48 Post Inoculation (PI) in the cloaca and cecum swabs and in litter samples and furthermore a reduction in CD8+ cells in the cecum compared to the challenged birds. Based on the results, it is concluded that feeding BS as a probiotic to broilers reduced the Salmonella spp. counts and thus, affected the mobilization of CD4+ and CD8+ cells in the ileum and cecum mucosa.

Este trabalho foi conduzido com o objetivo de avaliar a eficiência de um composto vegetal contendo óleo essencial de orégano, alecrim, canela e extrato de pimenta vermelha no controle de Salmonella, Eimeria e Clostridium em frangos de corte. Para tal, foram realizados dois experimentos. No primeiro avaliou-se a eficiência deste produto no controle de Clostridium perfringens após desafio com Eimeria acervulina, E. maxima e E. tenella. Aves de um dia de idade foram divididas em três grupos: T1 - dieta controle sem aditivo promotor de crescimento; T2 - dieta com adição de avilamicina (10ppm); e T3 - dieta com adição do composto vegetal (100ppm). O uso do composto vegetal na alimentação de frangos reduziu lesões específicas de E. maxima e E. tenella aos 14 dias pós-inoculação (PI) como também reduziram a contagem de unidades formadoras de colônias (UFC) de Clostridium perfringens no conteúdo do ceco das aves em relação ao grupo controle. No segundo experimento avaliou-se a eficiência deste mesmo produto em aves desafiadas com Salmonella Enteritidis. Aves de um dia de idade foram distribuídas em três tratamentos, sendo T1 - dieta controle sem adição de antibiótico promotor de crescimento, T2 - dieta com 10ppm de Avilamicina, T3 - dieta com 100ppm de um produto a base do composto vegetal acima citado. Aos 21 dias de idade todas as aves foram inoculadas com 10(5) UFC de Salmonella Enteritidis. A utilização do composto vegetal e avilamicina diminuiu a excreção de Salmonella nas aves 72 horas PI de Salmonella. A utilização do composto vegetal aumentou a relação vilo/células CD3+ no duodeno, em relação ao grupo avilamicina e controle, porém não teve efeito sobre a expressão destas células no ceco. The efficiency of a product in broiler feed containing essential oil of oregano, rosemary, cinnamon and extract of red pepper (plant compost) in the control of Salmonella, Eimeria and Clostridium was evaluated. Two experiments were carried out to evaluate the product. In the first experiment the efficiency of this product to control Clostridium perfringens after challenge with Eimeria acervulina, E. maxima and E. tenella was assessed. Day old chicks were allotted into three groups: T1 - control diet without growth promoter, T2 - diet with avilamycin (10ppm), and T3 - diet with addition of the plant compost (100ppm). The use of the plant compost in broiler diets reduced specific lesions of E. maxima and E. tenella at 14 days after inoculation and reduced the count of colony forming units (CFU) of Clostridium perfringens in the ceca comparing to the control group. In the second trial the efficiency of the same product in birds challenged with Salmonella Enteritidis was evaluated. Day old birds were submitted to three experimental diets: T1 - control diet without antibiotics growth promoter, T2 - diet with 10ppm Avilamycin, T3 - diet with 100ppm of the plant compost mentioned above. At 21 days of age all birds were inoculated with 10(5) CFU of Salmonella Enteritidis. The use of the plant compost and avilamycin decreased the excretion of Salmonella in poultry 72 hours after the inoculation. The use of the plant compost increased villous/CD3+ cells in the duodenum, compared to group avilamycin and control, but had no effect on the expression of these cells in the cecum.

Objetivou-se com o presente estudo comparar o efeito de diferentes sorovares de Salmonella na resposta imune local da mucosa do intestino de frangos de corte. Aos sete dias de idade, as aves foram desafiadas com os sorovares S. Enteritidis, S. Typhimurium, S. Senftenberg, S. Mbandaka e S. Minnesota. Foi observado que todos os sorovares testados foram capazes de colonizar o intestino das aves sendo possível o isolamento de Salmonella em suabes de cloaca, 48 h após inoculação. De maneira geral, as aves do grupo controle negativo, que não foram desafiados apresentaram quantidade significativamente menor de células imunológicas na mucosa intestinal do que as aves desafiadas. Porém, verificou-se que os sorovares de Salmonella, utilizados neste estudo, apresentaram diferentes efeitos sobre a dinâmica celular da mucosa do íleo e ceco e afetaram de modo diferente o ganho de peso e ganho médio diário das aves demonstrando distintos graus de patogenicidade. Os sorovares Enteritidis e Typhimurium apresentaram um efeito mais intenso tanto no desempenho quanto na mobilização de células imunológicas na mucosa intestinal de frangos de corte The study was designed to compare the effect of different Salmonella serovars in immune response across the count of CD8+ cells, CD4+ cells, goblet cells and macrophages in the gut mucosa of broilers. During the experimental inoculation at 7 day-old were used Salmonella enterica subspecies enterica sorovars Enteritidis, Typhimurium, Senftenberg, Mbandaka and Minnesota. It was observed that all serovars tested were capable of contaminating the poultry being possible counts of Salmonella in cloacal swabs, 48 h after inoculation and into the crop and cecum, at 14 and 20 day-old. Serovars tested had different effects on broiler performance assessed at 20 days. In the mucosa of the ileum and cecum of broilers, it was observed that some of the serotypes increased CD8 + cells, CD4 + cells, goblet cells and macrophages compared to the negative control group both at 14 and at 20 day-old. S. Enteritidis and S. Typhimurium are the serovars that showed the more intense effect in live performance and in the immune system of birds showing pathogenic characteristic; generally the broilers of the negative control showed significantly less immune cells on the intestinal mucosa than broilers inoculated experimentally. However, it was found that the Salmonella serovars used in this study had different effects on the cellular dynamics of the mucosa of the ileum and cecum and differently affect weight gain and average daily gain of poultry showing different levels of pathogenicity.

The present study was designed to compare the ability of different probiotics, applied in feed, to reduce Salmonella colonization and interfere with the cellular dynamics in the intestinal mucosa of broilers challenged with Salmonella enterica enterica sorovar Minnesota (SM). The following probiotics were used: A composed of Bacillus subtillis CCT 7711; B containing Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus rhamnosus, Lactobacillus bulgaricus, Enterococcus faecium, Streptococcus thermophilus, and Bifidobacterium bifidum; C consisting of Bacillus subtilis DSM 5750; and D with Enterococcus faecium. All tested probiotics were able to significantly reduce the Salmonella counts in cloacal swabs assessed at 48 h after inoculation when compared with the positive control. All probiotics were able to reduce the Salmonella counts in the cecum when compared to the positive control at 35 days. None of the tested probiotics reduced the Salmonella counts in the crop of broilers at 35 days of age. All probiotics induced an increase in the CD4 + and CD8+ cell counts in the cecum mucosa relative to the negative control group at 7 days of age; only probiotics C and D, reduced significantly the CD8 + cell counts in this same tissue of broilers at 35 days compared to positive control group. The results from this study indicated that the tested probiotics presented different effects on the reduction of SM counts and affected the cell dynamics in the cecum mucosa of broilers differently.

Dois experimentos foram desenvolvidos para avaliar a eficiência de ácidos orgânicos frente a Salmonella enterica enterica sorovar Enteritidis (SE) e Minnesota (SM) em frangos. No primeiro experimento foram avaliados 3 tratamentos: T1 - ração adicionada de ácido orgânico, T2 - ração adicionada de ácido orgânico e ácido orgânico na água de bebida, T3 - grupo controle. Todos os animais foram inoculados com SE, via oral. A utilização de ácidos orgânicos na ração (T1) e na ração e na água (T2) diminuíram a excreção de Salmonella no papo e no ceco 7 dias pós inoculação com SE e houve redução de células CD3+ no jejuno dos frangos. No segundo experimento foram avaliados 4 tratamentos sendo T1 - controle, T2 - controle inoculado via oral com Salmonella Minnesota (SM), T3 - animais inoculados via oral com SM e ácidos orgânicos na ração e T4 - animais inoculados via oral com SM e ácidos orgânicos na ração e na água de bebida. Ácidos orgânicos a ração (T3) e na ração e na água (T4) reduziram a excreção de SM em papo de frangos de corte desafiados, 7 dias após inoculação. O uso de ácidos orgânicos na ração e na ração e na água foram mais eficientes em reduzir SE do que SM.

Uma série de aditivos alimentares, incluindo antibióticos, tem sido amplamente utilizados na indústria avícola durante vários anos. A manipulação das funções gastrointestinais e da microbiota dos animais domésticos com aditivos alimentares é reconhecida como uma importante ferramenta para melhorar o desempenho e a eficiência alimentar. Recentemente, as preocupações com os possíveis resíduos de antibióticos e resistência a doenças tem despertado restrições no uso de antibióticos na indústria animal. A proibição do uso de antibióticos como aditivos na alimentação animal pela União Européia, tem acelerado as investigações sobre aditivos alternativos. Como alternativas, os óleos essenciais e ácidos orgânicos já são utilizados na alimentação animal para melhorar o desempenho e para controlar a população de microorganismos patogênicos no intestino em sistemas de manejo intensivo. O objetivo desta revisão é abordar a importância do equilíbrio da microbiota intestinal e como ácidos orgânicos e o carvacrol podem atuar no desenvolvimento da microbiota intestinal, no controle de bactérias patogênicas como Salmonella e Campylobacter e no desempenho zootécnico dos animais.