Immune Regulatory Effects of Enteromorphaclathrata Polysaccharides on Nd Attenuated Vaccine in a Chicken Model Infected with Reticuloendotheliosis Virus

Reticuloendotheliosis virus (REV) infection has frequently affected the poultry industry in recent years. The infection with REV weakens the immune responses of the infected poultry. It is reported that Enteromorphaclathrata polysaccharides are capable of regulating immune function. In order to investigate the immuno regulatory effects of Enteromorphaclathrata polysaccharides (EPS) on the response of REV-infected broilers to a live attenuated Newcastle disease (ND) vaccine. Broilers were intraperitoneally injected with REV at one day of age, subcutaneously infected with EPS at 2 days of age, and vaccinated by nasal drip with a live attenuated ND (Lasota strain) vaccine at 5 days of age. Immune organ index, secretory immunoglobulinA (SIgA), peripheral blood heterophil to lymphocyte ratios (H/L ratio), peripheral blood lymphocyte transformation rates, and interferon-gamma (IFN-) and interleukin-2 (IL-2) levels were measured at 3, 7, 14, 21, 28, 35, 42, and 56 days of age. The results showed that EPS increased the immune organ index, and the secretion of small intestine secretory immunoglobulin A, serum ND antibody titers, blood H/L ratio, peripheral blood lymphocyte transformation rates, and IL-2 and IFN- levels. These results indicate that EPS are able to enhance the immune responses of chickens both to REV infection and to ND vaccination. Therefore, Enteromorphaclathrata polysaccharides can be considered as an immune regulator in the future.(AU)

Immune Regulatory Effects of Enteromorphaclathrata Polysaccharides on Nd Attenuated Vaccine in a Chicken Model Infected with Reticuloendotheliosis Virus

Reticuloendotheliosis virus (REV) infection has frequently affected the poultry industry in recent years. The infection with REV weakens the immune responses of the infected poultry. It is reported that Enteromorphaclathrata polysaccharides are capable of regulating immune function. In order to investigate the immuno regulatory effects of Enteromorphaclathrata polysaccharides (EPS) on the response of REV-infected broilers to a live attenuated Newcastle disease (ND) vaccine. Broilers were intraperitoneally injected with REV at one day of age, subcutaneously infected with EPS at 2 days of age, and vaccinated by nasal drip with a live attenuated ND (Lasota strain) vaccine at 5 days of age. Immune organ index, secretory immunoglobulinA (SIgA), peripheral blood heterophil to lymphocyte ratios (H/L ratio), peripheral blood lymphocyte transformation rates, and interferon-gamma (IFN-) and interleukin-2 (IL-2) levels were measured at 3, 7, 14, 21, 28, 35, 42, and 56 days of age. The results showed that EPS increased the immune organ index, and the secretion of small intestine secretory immunoglobulin A, serum ND antibody titers, blood H/L ratio, peripheral blood lymphocyte transformation rates, and IL-2 and IFN- levels. These results indicate that EPS are able to enhance the immune responses of chickens both to REV infection and to ND vaccination. Therefore, Enteromorphaclathrata polysaccharides can be considered as an immune regulator in the future.

Protection of chickens against fowl typhoid using field vaccine programs formulated with the live attenuated strain Salmonella Gallinarum deltacobSdeltacbiA / Proteção de aves contra o tifo aviário utilizando a cepa atenuada Salmonella Gallinarum deltacobSdeltacbiA em diferentes programas vacinais de campo

Salmonella enterica serovar Gallinarum biovar Gallinarum (SG) is a host-specific bacteria that causes the fowl typhoid (FT). This disease is highly pathogenic to commercial chickens, specially brown layers and breeders, causing acute septicemia followed by high morbidity and mortality. Vaccination is extensively adopted in the fields as a biosafety tool for prevention of isolated infections and outbreaks in commercial poultry flocks. The present study evaluated the use of an attenuated SG with deletions on genes cobS and cbiA (SGΔcobSΔcbiA) as a live vaccine, using vaccination schemes adjusted for field conditions. To this end, brown layers were used in two different experiments, to evaluate the long-term protection, necessary in the fields. The vaccination scheme on the first experiment consisted of two doses, the first at 4 th week-of-age and the booster dose at 8 th week-of-age with challenge at 16 th week-of-age with wild SG strain. On the second experiment, the vaccination was carried out by different routes using three doses of the live vaccine, at 4 th , 8 th and 12 th weeks-of-age, and the challenge was done at 20 th weeks-of-age. After the challenge, the mortality was recorded during 28 days, and the egg production (experiment 2) was evaluated and compared with the group of unvaccinated layers. In both experiments, the mortality was significantly reduced, and the egg production was not affected in vaccinated layer-hens. In summary, this study shows the efficacy and the protection of different vaccination schemes against FT that can be applied under field conditions in commercial poultry farms.

Salmonella enterica sorovar Gallinarum biovar Gallinarum (SG) é uma bactéria hospedeira específica que causa o tifo aviário (TA). Essa doença é altamente patogênica em aves comerciais, especialmente galinhas poedeiras de linhagem vermelha e aves reprodutoras pesadas, causando septicemia aguda, e consequentemente, alta morbidade e mortalidade. A vacinação é amplamente utilizada no campo como uma ferramenta de biossegurança para a prevenção de infecções isoladas e surtos nas granjas avícolas comerciais. O atual estudo avaliou o potencial vacinal de uma cepa viva atenuada de SG com deleções nos genes cobS e cbiA (SGΔcobSΔcbiA), utilizando esquemas de vacinação formulados para utilização em campo. Para isso, as galinhas poedeiras de linhagem vermelha foram utilizadas em dois experimentos diferentes, para avaliar a proteção a longo prazo, necessária no campo. O esquema de vacinação no primeiro experimento consistiu em duas doses, a primeira na quarta semana de vida e a dose de reforço na oitava, e o desafio na 16ª semana com a estirpe selvagem SG. No segundo experimento, a vacinação foi realizada por diferentes rotas usando três doses da vacina viva, na quarta, na oitava e na décima segunda semana de vida, e o desafio foi feito na 20ª semana de vida. Após o desafio, a mortalidade foi acompanhada por 28 dias, e no experimento 2 a produção de ovos também foi avaliada e comparada com o grupo de galinhas não vacinadas. Em ambos os experimentos, a mortalidade foi significativamente reduzida, e a produção de ovos não foi afetada nos grupos de galinhas poedeiras vacinadas. Este estudo mostra a eficácia da proteção dos diferentes programas de vacinação contra o TA, que podem ser aplicados em granjas comerciais em condições de campo.

Protection of chickens against fowl typhoid using field vaccine programs formulated with the live attenuated strain Salmonella Gallinarum ΔcobSΔcbiA / Proteção de aves contra o tifo aviário utilizando a cepa atenuada Salmonella Gallinarum ΔcobSΔcbiA em diferentes programas vacinais de campo

Salmonella enterica serovar Gallinarum biovar Gallinarum (SG) is a host-specific bacteria that causes the fowl typhoid (FT). This disease is highly pathogenic to commercial chickens, specially brown layers and breeders, causing acute septicemia followed by high morbidity and mortality. Vaccination is extensively adopted in the fields as a biosafety tool for prevention of isolated infections and outbreaks in commercial poultry flocks. The present study evaluated the use of an attenuated SG with deletions on genes cobS and cbiA (SGΔcobSΔcbiA) as a live vaccine, using vaccination schemes adjusted for field conditions. To this end, brown layers were used in two different experiments, to evaluate the long-term protection, necessary in the fields. The vaccination scheme on the first experiment consisted of two doses, the first at 4 th week-of-age and the booster dose at 8 th week-of-age with challenge at 16 th week-of-age with wild SG strain. On the second experiment, the vaccination was carried out by different routes using three doses of the live vaccine, at 4 th , 8 th and 12 th weeks-of-age, and the challenge was done at 20 th weeks-of-age. After the challenge, the mortality was recorded during 28 days, and the egg production (experiment 2) was evaluated and compared with the group of unvaccinated layers. In both experiments, the mortality was significantly reduced, and the egg production was not affected in vaccinated layer-hens. In summary, this study shows the efficacy and the protection of different vaccination schemes against FT that can be applied under field conditions in commercial poultry farms.(AU)

Salmonella enterica sorovar Gallinarum biovar Gallinarum (SG) é uma bactéria hospedeira específica que causa o tifo aviário (TA). Essa doença é altamente patogênica em aves comerciais, especialmente galinhas poedeiras de linhagem vermelha e aves reprodutoras pesadas, causando septicemia aguda, e consequentemente, alta morbidade e mortalidade. A vacinação é amplamente utilizada no campo como uma ferramenta de biossegurança para a prevenção de infecções isoladas e surtos nas granjas avícolas comerciais. O atual estudo avaliou o potencial vacinal de uma cepa viva atenuada de SG com deleções nos genes cobS e cbiA (SGΔcobSΔcbiA), utilizando esquemas de vacinação formulados para utilização em campo. Para isso, as galinhas poedeiras de linhagem vermelha foram utilizadas em dois experimentos diferentes, para avaliar a proteção a longo prazo, necessária no campo. O esquema de vacinação no primeiro experimento consistiu em duas doses, a primeira na quarta semana de vida e a dose de reforço na oitava, e o desafio na 16ª semana com a estirpe selvagem SG. No segundo experimento, a vacinação foi realizada por diferentes rotas usando três doses da vacina viva, na quarta, na oitava e na décima segunda semana de vida, e o desafio foi feito na 20ª semana de vida. Após o desafio, a mortalidade foi acompanhada por 28 dias, e no experimento 2 a produção de ovos também foi avaliada e comparada com o grupo de galinhas não vacinadas. Em ambos os experimentos, a mortalidade foi significativamente reduzida, e a produção de ovos não foi afetada nos grupos de galinhas poedeiras vacinadas. Este estudo mostra a eficácia da proteção dos diferentes programas de vacinação contra o TA, que podem ser aplicados em granjas comerciais em condições de campo.(AU)

Protection of chickens against fowl typhoid using field vaccine programs formulated with the live attenuated strain Salmonella Gallinarum deltacobSdeltacbiA / Proteção de aves contra o tifo aviário utilizando a cepa atenuada Salmonella Gallinarum deltacobSdeltacbiA em diferentes programas vacinais de campo

Salmonella enterica serovar Gallinarum biovar Gallinarum (SG) is a host-specific bacteria that causes the fowl typhoid (FT). This disease is highly pathogenic to commercial chickens, specially brown layers and breeders, causing acute septicemia followed by high morbidity and mortality. Vaccination is extensively adopted in the fields as a biosafety tool for prevention of isolated infections and outbreaks in commercial poultry flocks. The present study evaluated the use of an attenuated SG with deletions on genes cobS and cbiA (SGΔcobSΔcbiA) as a live vaccine, using vaccination schemes adjusted for field conditions. To this end, brown layers were used in two different experiments, to evaluate the long-term protection, necessary in the fields. The vaccination scheme on the first experiment consisted of two doses, the first at 4 th week-of-age and the booster dose at 8 th week-of-age with challenge at 16 th week-of-age with wild SG strain. On the second experiment, the vaccination was carried out by different routes using three doses of the live vaccine, at 4 th , 8 th and 12 th weeks-of-age, and the challenge was done at 20 th weeks-of-age. After the challenge, the mortality was recorded during 28 days, and the egg production (experiment 2) was evaluated and compared with the group of unvaccinated layers. In both experiments, the mortality was significantly reduced, and the egg production was not affected in vaccinated layer-hens. In summary, this study shows the efficacy and the protection of different vaccination schemes against FT that can be applied under field conditions in commercial poultry farms.(AU)

Salmonella enterica sorovar Gallinarum biovar Gallinarum (SG) é uma bactéria hospedeira específica que causa o tifo aviário (TA). Essa doença é altamente patogênica em aves comerciais, especialmente galinhas poedeiras de linhagem vermelha e aves reprodutoras pesadas, causando septicemia aguda, e consequentemente, alta morbidade e mortalidade. A vacinação é amplamente utilizada no campo como uma ferramenta de biossegurança para a prevenção de infecções isoladas e surtos nas granjas avícolas comerciais. O atual estudo avaliou o potencial vacinal de uma cepa viva atenuada de SG com deleções nos genes cobS e cbiA (SGΔcobSΔcbiA), utilizando esquemas de vacinação formulados para utilização em campo. Para isso, as galinhas poedeiras de linhagem vermelha foram utilizadas em dois experimentos diferentes, para avaliar a proteção a longo prazo, necessária no campo. O esquema de vacinação no primeiro experimento consistiu em duas doses, a primeira na quarta semana de vida e a dose de reforço na oitava, e o desafio na 16ª semana com a estirpe selvagem SG. No segundo experimento, a vacinação foi realizada por diferentes rotas usando três doses da vacina viva, na quarta, na oitava e na décima segunda semana de vida, e o desafio foi feito na 20ª semana de vida. Após o desafio, a mortalidade foi acompanhada por 28 dias, e no experimento 2 a produção de ovos também foi avaliada e comparada com o grupo de galinhas não vacinadas. Em ambos os experimentos, a mortalidade foi significativamente reduzida, e a produção de ovos não foi afetada nos grupos de galinhas poedeiras vacinadas. Este estudo mostra a eficácia da proteção dos diferentes programas de vacinação contra o TA, que podem ser aplicados em granjas comerciais em condições de campo.(AU)

Rattlesnake Crotalus molossus nigrescens venom induces oxidative stress on human erythrocytes

Background Globally, snake envenomation is a well-known cause of death and morbidity. In many cases of snakebite, myonecrosis, dermonecrosis, hemorrhage and neurotoxicity are present. Some of these symptoms may be provoked by the envenomation itself, but others are secondary effects of the produced oxidative stress that enhances the damage produced by the venom toxins. The only oxidative stress effect known in blood is the change in oxidation number of Fe (from ferrous to ferric) in hemoglobin, generating methemoglobin but not in other macromolecules. Currently, the effects of the overproduction of methemoglobin derived from snake venom are not extensively recorded. Therefore, the present study aims to describe the oxidative stress induced by Crotalus molossus nigrescens venom using erythrocytes. Methods Human erythrocytes were washed and incubated with different Crotalus molossus nigrescens venom concentrations (0-640 μg/mL). After 24 h, the hemolytic activity was measured followed by attenuated total reflectance-Fourier transform infrared spectroscopy, non-denaturing PAGE, conjugated diene and thiobarbituric acid reactive substances determination. Results Low concentrations of venom (<10 μg/mL) generates oxyhemoglobin release by hemolysis, whereas higher concentrations produced a hemoglobin shift of valence, producing methemoglobin (>40 μg/mL). This substance is not degraded by proteases present in the venom. By infrared spectroscopy, starting in 80 μg/mL, we observed changes in bands that are associated with protein damage (1660 and 1540 cm−1) and lipid peroxidation (2960, 2920 and 1740 cm−1). Lipid peroxidation was confirmed by conjugated diene and thiobarbituric acid reactive substance determination, in which differences were observed between the control and erythrocytes treated with venom. Conclusions Crotalus molossus nigrescens venom provokes hemolysis and oxidative stress, which induces methemoglobin formation, loss of protein structure and lipid peroxidation.(AU)

Rattlesnake Crotalus molossus nigrescens venom induces oxidative stress on human erythrocytes

Background Globally, snake envenomation is a well-known cause of death and morbidity. In many cases of snakebite, myonecrosis, dermonecrosis, hemorrhage and neurotoxicity are present. Some of these symptoms may be provoked by the envenomation itself, but others are secondary effects of the produced oxidative stress that enhances the damage produced by the venom toxins. The only oxidative stress effect known in blood is the change in oxidation number of Fe (from ferrous to ferric) in hemoglobin, generating methemoglobin but not in other macromolecules. Currently, the effects of the overproduction of methemoglobin derived from snake venom are not extensively recorded. Therefore, the present study aims to describe the oxidative stress induced by Crotalus molossus nigrescens venom using erythrocytes. Methods Human erythrocytes were washed and incubated with different Crotalus molossus nigrescens venom concentrations (0-640 μg/mL). After 24 h, the hemolytic activity was measured followed by attenuated total reflectance-Fourier transform infrared spectroscopy, non-denaturing PAGE, conjugated diene and thiobarbituric acid reactive substances determination. Results Low concentrations of venom (<10 μg/mL) generates oxyhemoglobin release by hemolysis, whereas higher concentrations produced a hemoglobin shift of valence, producing methemoglobin (>40 μg/mL). This substance is not degraded by proteases present in the venom. By infrared spectroscopy, starting in 80 μg/mL, we observed changes in bands that are associated with protein damage (1660 and 1540 cm−1) and lipid peroxidation (2960, 2920 and 1740 cm−1). Lipid peroxidation was confirmed by conjugated diene and thiobarbituric acid reactive substance determination, in which differences were observed between the control and erythrocytes treated with venom. Conclusions Crotalus molossus nigrescens venom provokes hemolysis and oxidative stress, which induces methemoglobin formation, loss of protein structure and lipid peroxidation.(AU)

Rattlesnake Crotalus molossus nigrescens venom induces oxidative stress on human erythrocytes

Abstract Background Globally, snake envenomation is a well-known cause of death and morbidity. In many cases of snakebite, myonecrosis, dermonecrosis, hemorrhage and neurotoxicity are present. Some of these symptoms may be provoked by the envenomation itself, but others are secondary effects of the produced oxidative stress that enhances the damage produced by the venom toxins. The only oxidative stress effect known in blood is the change in oxidation number of Fe (from ferrous to ferric) in hemoglobin, generating methemoglobin but not in other macromolecules. Currently, the effects of the overproduction of methemoglobin derived from snake venom are not extensively recorded. Therefore, the present study aims to describe the oxidative stress induced by Crotalus molossus nigrescens venom using erythrocytes. Methods Human erythrocytes were washed and incubated with different Crotalus molossus nigrescens venom concentrations (0640 g/mL). After 24 h, the hemolytic activity was measured followed by attenuated total reflectance-Fourier transform infrared spectroscopy, non-denaturing PAGE, conjugated diene and thiobarbituric acid reactive substances determination. Results Low concentrations of venom ( 10 g/mL) generates oxyhemoglobin release by hemolysis, whereas higher concentrations produced a hemoglobin shift of valence, producing methemoglobin (>40 g/mL). This substance is not degraded by proteases present in the venom. By infrared spectroscopy, starting in 80 g/mL, we observed changes in bands that are associated with protein damage (1660 and 1540 cm1) and lipid peroxidation (2960, 2920 and 1740 cm1). Lipid peroxidation was confirmed by conjugated diene and thiobarbituric acid reactive substance determination, in which differences were observed between the control and erythrocytes treated with venom. Conclusions Crotalus molossus nigrescens venom provokes hemolysis and oxidative stress, which induces methemoglobin formation, loss of protein structure and lipid peroxidation.

ASSOCIAÇÃO DE VACINA CONTRA COCCIDIOSE E ADITIVO NUTRICIONAL EM FRANGOS DE CORTE DESAFIADOS EXPERIMENTALMENTE

O objetivo deste estudo foi avaliar a eficácia de uma vacina atenuada por precocidade como alternativa as drogas anticoccidianas utilizadas no controle da coccidiose, associado a um aditivo nutricional na dieta, sobre o desempenho produtivo e a saúde intestinal de frangos de corte desafiados. Foram utilizados 672 pintinhos machos, da linhagem Ross, provenientes de matrizes de cerca de 40 semanas de idade. As aves foram distribuídas em um delineamento inteiramente casualizado em esquema fatorial 4x2 (4 programas de controle vs 2 condições sanitárias) totalizando 8 tratamentos com 6 repetições de 14 aves cada. Os resultados obtidos no experimento foram analisados utilizando-se análise de variância (ANOVA) do procedimento General Lineal Model (GLM) com auxílio do programa estatístico SAS (Statistical Analysis System) e quando significativas, as médias entre os tratamentos foram comparadas pelo teste Tukey. As aves que foram desafiadas apresentaram menores (P<0,05) resultados de desempenho produtivo independente dos tratamentos, demonstrando similaridade na efetividade de ambos os programas anticoccidianos. A associação de vacina e aditivo melhorou (P<0,05) a conversão alimentar entre 21 e 28 dias. Para morfometria do duodeno, houve menor (P<0,05) relação vilo:cripta aos 19 dias e área de absorção aos 28 dias para grupo desafiado. No desdobramento da interação, o programa anticoccidiano acrescido de aditivo resultou em menor (P<0,05) espessura da camada muscular do grupo desafiado. No jejuno, aves vacinadas demonstraram maior (P<0,05) largura de vilo e o programa anticoccdiano maior (P<0,05) área de absorção aos 19 dias. No desdobramento da interação, a associação de aditivo e vacina melhorou (P<0,05) a profundidade de cripta e o programa vacina resultou em menor (P<0,05) relação vilo:cripta para aves desafiadas. Aos 28 dias, o programa vacina apresentou maior (P<0,05) comprimento do vilo e profundidade de cripta, o que demonstra maior resposta regenerativa do jejuno. No íleo, a associação de vacina e aditivo apresentou maior (P<0,05) largura de vilo aos 19 dias e o programa anticoccidano acrescido de aditivo apresentou menor (P<0,05) profundidade de cripta e maior área de absorção aos 28 dias, porém não apresentou melhores valores que o programa vacina acrescido de aditivo. Aves desafiadas apresentaram maior (P<0,05) distância de deslocamente do probe aos 19 dias, ou seja, característica mais elástica, pois no momento da cicatrização do tecido lesionado é utilizado colágeno. Não houve (P>0,05) efeito significativo para oxidação lipídica, contagem de células em mitose na cripta da mucosa do jejuno em relação aos programas de controle e as condições sanitárias avaliadas. O programa vacina anticoccidiana acrescido de aditivo apresentou maior (P<0,05) capacidade antioxidante sérica de DPPH aos 19 dias. O grupo vacinado demonstrou o maior nível de ABTS aos 19 dias, impactando positivamente na capacidade de proteção antioxidante. Os níveis séricos de FITC-d foram menores (P<0,05) para grupo desafiado. No desdobramento da interação dos fatores para o valor b* mensurado na pele do peito das aves, observou-se maior (P<0,05) valor quando foi adicionado o aditivo nutricional ao programa vacina. Aves vacinadas demonstraram menor escore de lesão para E. tenella aos 19 dias. O programa vacina associado ao aditivo, resultou em maior (P<0,05) coloração de pés para grupo controle aos 19 dias comparado com o programa anticoccidiano na ausência de aditivo. O comportamento de excreção de oocistos das aves vacinadas demonstrou formação de imunidade e ambos os programas anticoccidianos foram efetivos para controle da coccidiose. O controle da coccidiose em frangos de corte com uso de vacina atenuada por precocidade pode ser uma alternativa eficaz e segura e capaz de proporcionar desempenho zootécnico semelhante àquele obtido com o programa anticoccidiano em situações de desafio ou não. O uso de um aditivo nutricional com ácidos graxos de cadeia curta e média pode ser usado em associação com programa anticoccidiano para potencializar os resultados.

This research aimed to avaluate the effectiveness of a precocity attenuated vaccine as an alternative to anticoccidial drugs used to control coccidiosis, associated with a nutritional additive in the diet, on the productive performance and the intestinal health of challenged broilers. For the accomplishment of this study 672 male chicks of the Cobb Slow line with from dams around 40 weeks old. The birds were distributed in a completely randomized design in a 4 x 2 factorial scheme (4 control programs versus 2 sanitary conditions) resulting in 8 treatments and 6 replicates of 14 birds per box. The results obtained in the experiment were analyzed using analysis of variance (ANOVA) of the General Lineal Model (GLM) procedure with the aid of the statistical program SAS (Statistical Analysis System) and when significant, the means between treatments were compared byTukey test. The birds that were challenged had smaller (P<0,05) performance results regardless of the of treatments, demonstrating similarity in the effectiveness of the both anticoccidial programs. The association of vaccine and additive improved (P<0,05) feed conversion between 21 and 28 days. For duodenum morphometry, there was smaller (P<0,05) villus:crypt ratio at 19 days and absorption area at 28 days for the challenged group. In the unfolding of the interaction, the anticoccidial program with the addition of additive resulted in smaller (P<0,05) muscle layer thickness in the challenged group. In the jejunum, vaccinated birds demonstrated greater (P<0,05) villi width and the anticoccidial program greater area of absorption at 19 days. In the unfolding of the interaction, the association of vaccine and additive improved (P<0,05) depth of the crypt and the vaccine program resulted in smaller (P<0,05) vilo:crypt ratio for challenged birds. At 28 days, the vaccine program presented greater (P<0,05) villus length and crypt depth, which demonstrates higher jejunum regenerative response. In the ileus, the association of vaccine and additive showed greater (P<0,05) villi width at 19 days and the anticoccidial program with the addition of additive presented smaller (P<0,05) crypt depth and greater area of absorption at 28 days, however it did not show better values than the vaccine program plus an additive. Challenged birds had a greater (P<0.05) probe displacement distance at 19 days, that is, a more elastic characteristic, as collagen is used at the time of healing of the injured tissue. There was no significant effect (P>0,05) for lipid oxidation and cell count in mitosis in the jejunal mucosa crypt in relation to the control programs and the sanitary conditions evaluated. The vaccine program plus additive showed higher serum antioxidant capacity of DPPH at 19 days. The vaccinated group showed the highest (P<0,05) level of ABTS at 19 days, positively impacting the antioxidant protection capacity. Serum FITC-d levels were lower (P>0.05) for the challenged group. In the unfolding of the interaction of the factors for the b* value measured on the broiler breast skin, a greater (P<0,05) value was observed when the nutritional additive was added to the vaccine program. The vaccinated chickens showed smaller (P<0.05) lesion score for E. tenella at 19 days. The vaccine program associated with the additive resulted in greater coloration of chicken feet for the control group at 19 days compared to the anticoccidial program without additive inclusion. The oocyst excretion behavior of the vaccinated birds demonstrated immunity formation and both anticoccidial programs were effective for the control of coccidiosis. The control of coccidiosis in broiler chickens using a vaccine attenuated by precocity can be an effective and safe alternative, capable of providing zootechnical performance similar to that obtained with the anticoccidial program in challenging situations or not. The use of a nutritional additive with short and medium chain fatty acids can be used in association with an anticoccidial program to enhance the results.

A hematologic and electrophoretic study in puppies vaccinated against canine distemper virus and canine parvovirus

Background: Canine distemper is a contagious multisystemic viral disease that affects canines and others carnivores. Canine parvovirus infection is one of the most important viral diseases in young dogs. Side effects of vaccine generally include fever, lethargy and local inflammation. Complementary exams are important to evaluate the strenght of immungenic stimulation. This study was aimed at evaluating hematological and electrophoretic alterations in puppies after inoculation of live attenuated vaccine against canine distemper virus and canine parvovirus. Materials, Methods & Results: Five non-breeding newborn dogs of the same litter were used. Animals received three subcutaneous injection of 1mL (at days 0, 21 and 42). Blood was collected at day 0 (day of vaccination) and for three times for each dose: at days 7, 14 and 21 (first dose); at days 28, 35 and 42 (second dose); and at days 49, 56 and 63 (third dose). Blood containing ethylenediaminetetraacetic acid as anticoagulant was used for hematological evaluation. The total serum protein were determined by the biuret method, using commercial reagent, according to fabricant instructions. Serum was used for protein fractionation by using cellulose acetate strip electrophoresis. A decrease in platelet count was observed at days 7 and 28 post-vaccination. Lymphocyte number increased 88.4%, as well as the level of the protein fractions alpha-1 globulin (68%) and alpha-2 globulin (41.4%) at day 7. Moreover, a 5-fold increase in the fibrinogem concentration and in the number of eosinophils was observed at day 14. Thereafter, the platelet count decreased by 27.3% and the number of monocytes increased 5-fold at day 28. Discussion: Mild to moderate thrombocytopenia is often observed in dogs 3-5 days post-vaccination with live attenuated vaccines, mainly those against CDV and CPV. Besides the platelet damage caused by the CDV per se, infected animals showed secondary immune-mediated thrombocytopenia and decreased platelet production due to direct viral megakaryocyte infection. The increase in alpha-1 globulin may be related to the augment in the synthesis of alpha-1 antitrypsin, the main protein of the alpha-1 globulin region, in response to the vaccine-induced acute inflammatory process. The alpha-2 globulin region includes haptoglobin, alpha-2 macroglobulin and ceruloplasmin, and the increase observed in this fraction suggested that both haptoglobin and ceruloplasmin levels were augmented, following acute inflammatory response pattern. Fibrinogen is a soluble plasma glycoprotein that is converted by thrombin into fibrin during blood clotting. Despite the increase in fibrinogen concentration be the best indicator of inflammation in large animals, the hyperfibrinogenemia observed suggests that the inflammatory process was adequate to stimulate synthesis of this acute phase protein (P < 0,05). Absence of lymphocytosis observed at days 49, 56 and 63 associated to the progressive increase of the gamma globulin fraction, although not statistically significant, suggested an augment of B lymphocytes. The eosinophilia was observed in highlighting the presence of inflammation. Moreover, an increase in monocyte count indicating the presence of subacute or chronic inflammation after the second dose of the vaccine.

A hematologic and electrophoretic study in puppies vaccinated against canine distemper virus and canine parvovirus / A hematologic and electrophoretic study in puppies vaccinated against canine distemper virus and canine parvovirus

Background: Canine distemper is a contagious multisystemic viral disease that affects canines and others carnivores. Canine parvovirus infection is one of the most important viral diseases in young dogs. Side effects of vaccine generally include fever, lethargy and local inflammation. Complementary exams are important to evaluate the strenght of immungenic stimulation. This study was aimed at evaluating hematological and electrophoretic alterations in puppies after inoculation of live attenuated vaccine against canine distemper virus and canine parvovirus.Materials, Methods & Results: Five non-breeding newborn dogs of the same litter were used. Animals received three subcutaneous injection of 1mL (at days 0, 21 and 42). Blood was collected at day 0 (day of vaccination) and for three times for each dose: at days 7, 14 and 21 (first dose); at days 28, 35 and 42 (second dose); and at days 49, 56 and 63 (third dose). Blood containing ethylenediaminetetraacetic acid as anticoagulant was used for hematological evaluation. The total serum protein were determined by the biuret method, using commercial reagent, according to fabricant instructions. Serum was used for protein fractionation by using cellulose acetate strip electrophoresis. A decrease in platelet count was observed at days 7 and 28 post-vaccination. Lymphocyte number increased 88.4%, as well as the level of the protein

Background: Canine distemper is a contagious multisystemic viral disease that affects canines and others carnivores. Canine parvovirus infection is one of the most important viral diseases in young dogs. Side effects of vaccine generally include fever, lethargy and local inflammation. Complementary exams are important to evaluate the strenght of immungenic stimulation. This study was aimed at evaluating hematological and electrophoretic alterations in puppies after inoculation of live attenuated vaccine against canine distemper virus and canine parvovirus.Materials, Methods & Results: Five non-breeding newborn dogs of the same litter were used. Animals received three subcutaneous injection of 1mL (at days 0, 21 and 42). Blood was collected at day 0 (day of vaccination) and for three times for each dose: at days 7, 14 and 21 (first dose); at days 28, 35 and 42 (second dose); and at days 49, 56 and 63 (third dose). Blood containing ethylenediaminetetraacetic acid as anticoagulant was used for hematological evaluation. The total serum protein were determined by the biuret method, using commercial reagent, according to fabricant instructions. Serum was used for protein fractionation by using cellulose acetate strip electrophoresis. A decrease in platelet count was observed at days 7 and 28 post-vaccination. Lymphocyte number increased 88.4%, as well as the level of the protein

Post-challenge hematological evaluation with virulent strain of Eimeria tenella in broilers immunized with attenuated strain or sporozoite proteins from homologous strain

Hematological parameters were evaluated in broilers immunized and challenged with Eimeria tenella. Broiler chickens of Hubbard strain, females, coccidian-free, were kept in wire cages and inoculated on the third day. The experiment was designed to include five sorts of treatment with three replicates each. T1 was the negative control group, T2 received 500 attenuated sporulated oocysts by gavage, T3 was the positive control, T4 received 50 µg of sporozoite protein + Quil A vaccine, and T5 received Quil A without sporozoite protein + PBS, the last two through nasal route on days 0, 7, and 21. On the 31st day, all treatments were challenged with homologous virulent strain of E. tenella in the dose of 8.0 × 10(4) oocysts, with the exception of T1. One week later, blood sampling, lesion scores, and cecal oocyst count were carried out. The parasitological parameters showed statistical significance (p 0.05) and there was no damage to the hematological parameters of birds (p > 0.05) by ANOVA test. The correlations suggest that the blood parameters were impaired by effects of the parasite on tissue, showing levels of hemorrhage and/or hydration.

Foram avaliados os parâmetros hematológicos em frangos de corte imunizados e desafiados com Eimeria tenella. Pintos de corte fêmeas da linhagem Hubbard, livres de coccídios, foram mantidos em baterias metálicas e inoculados no terceiro dia. O experimento foi delineado por cinco tratamentos com três repetições cada, sendo: T1 controle negativo, T2 recebeu 500 oocistos esporulados atenuados via oral, T3 controle positivo, T4 recebeu vacina contendo 50 µg de proteínas de esporozoítos + Quil A e T5 recebeu Quil A + PBS, sendo os dois últimos por via nasal nos dias 0, 7 e 21. No dia 31, todos os tratamentos foram desafiados com cepa virulenta homóloga de E. tenella na dose de 8,0 × 10(4) oocistos, exceto T1. Uma semana depois, foi realizada amostragem de sangue, escore de lesão e contagem de oocistos cecais. Os parâmetros parasitológicos apresentaram significância estatística (p 0,05), sem que causassem prejuízos aos parâmetros hematológicos das aves (p > 0,05), pelo teste ANOVA. As correlações sugerem que os parâmetros sanguíneos foram afetados pelos efeitos do parasita no tecido, apresentando níveis de hemorragia e/ou hidratação.

Effect of maternally-derived antibodies on the performance and immunity of broilers induced by in ovo or post-hatching immunizations with a live vaccine against infectious bursal disease

The interference of low or high maternal antibodies titers on the attenuated infectious bursal disease (IBD) virus (IBDV) vaccine infection and its effects on the performance of broilers vaccinated at the 18th day of incubation (in ovo), at one day of age (subcutaneously-SC), or at 15 days of age (drinking water-DW) were investigated. After a series of three live vaccinations, breeders were given or not an IBD oil emulsion vaccine (IBD-OEV) prior to sexual maturity. At day 18 of incubation (in ovo), a commercial vaccine containing HVT and an intermediate IBDV strain or the single HVT vaccine was given. An intermediate IBDV vaccine was given SC at one day of age, or at 15 days of age via DW. The progeny of unvaccinated breeders presented higher neutralizing IBDVspecific antibody (IBDVab) titers at 25 and 40 days of age than those of the progeny of IBD-OEV breeders (p 0.05) at any broilers vaccination age and route. The lower IBDV RNA detection by RT-PCR in the bursa of Fabricius (BF) and the lower IBDV antibody titers in the serum of the groups vaccinated at one and 15 days of age derived from IBD-OEV breeders may indicate antibody-mediated IBDV neutralization. The inovo and one-day vaccinations did not interfere with performance, both in low and high antibody-titered progenies. The in-ovo vaccination against IBD is considered convenient and safe for industrial chickens, irrespective their maternal antibody levels.

Effect of maternally-derived antibodies on the performance and immunity of broilers induced by in ovo or post-hatching immunizations with a live vaccine against infectious bursal disease

The interference of low or high maternal antibodies titers on the attenuated infectious bursal disease (IBD) virus (IBDV) vaccine infection and its effects on the performance of broilers vaccinated at the 18th day of incubation (in ovo), at one day of age (subcutaneously-SC), or at 15 days of age (drinking water-DW) were investigated. After a series of three live vaccinations, breeders were given or not an IBD oil emulsion vaccine (IBD-OEV) prior to sexual maturity. At day 18 of incubation (in ovo), a commercial vaccine containing HVT and an intermediate IBDV strain or the single HVT vaccine was given. An intermediate IBDV vaccine was given SC at one day of age, or at 15 days of age via DW. The progeny of unvaccinated breeders presented higher neutralizing IBDVspecific antibody (IBDVab) titers at 25 and 40 days of age than those of the progeny of IBD-OEV breeders (p 0.05) at any broilers vaccination age and route. The lower IBDV RNA detection by RT-PCR in the bursa of Fabricius (BF) and the lower IBDV antibody titers in the serum of the groups vaccinated at one and 15 days of age derived from IBD-OEV breeders may indicate antibody-mediated IBDV neutralization. The inovo and one-day vaccinations did not interfere with performance, both in low and high antibody-titered progenies. The in-ovo vaccination against IBD is considered convenient and safe for industrial chickens, irrespective their maternal antibody levels.

A defective mutant of Salmonella enterica Serovar Gallinarum in cobalamin biosynthesis is avirulent in chickens

Salmonella enterica serovar Gallinarum (SG) is a fowl typhoid agent in chickens and is a severe disease with worldwide economic impact as its mortality may reach up to 80%. It is one of a small group of serovars that typically produces typhoid-like infections in a narrow range of host species and which therefore represents a good model for human typhoid. The survival mechanisms are not considered to be virulent mechanisms but are essential for the life of the bacterium. Mutants of Salmonella Gallinarum containing defective genes, related to cobalamin biosynthesis and which Salmonella spp. has to be produced to survive when it is in an anaerobic environment, were produced in this study. Salmonella Gallinarum is an intracellular parasite. Therefore, this study could provide information about whether vitamin B12 biosynthesis might be essential to its survival in the host. The results showed that the singular deletion in cbiA or cobS genes did not interfere in the life of Salmonella Gallinarum in the host, perhaps because single deletion is not enough to impede vitamin B12 biosynthesis. It was noticed that diluted SG mutants with single deletion produced higher mortality than the wild strain of SG. When double mutation was carried out, the Salmonella Gallinarum mutant was unable to provoke mortality in susceptible chickens. This work showed that B12 biosynthesis is a very important step in the metabolism of Salmonella Gallinarum during the infection of the chickens. Further research on bacterium physiology should be carried out to elucidate the events described in this research and to assess the mutant as a vaccine strain.

Salmonella enterica serovar Gallinarum (SG) é o agente do tifo aviário, doença severa que provoca mortalidade em até 80% do plantel de aves. SG encontra-se entre os poucos sorotipos de Salmonella que são agentes etiológicos de enfermidade específica, à semelhança de Salmonella Typhi em seres humanos podendo, portanto, servir de modelo experimental para outras salmoneloses hospedeiro especíifcas. Além dos mecanismos de virulência, a bactéria utiliza mecanismos de sobrevivência para permanecer no hospedeiro. A ativação desses mecanismos pode ou não estar associada à ativação dos mecanismos de virulência. Entre os mecanismos fisiológicos, está a produção de vitamina B12 que Salmonella spp. realiza em ambientes anaeróbicos, como quando encontra-se intracelularmente no organismo hospedeiro. Neste estudo, analisou-se a infecção de aves por cepas de SG, que tiveram genes alterados que participam da biossíntese de vitamina B12. Foram produzidos mutantes de SG contendo os genes cbiA e cobS alterados e um terceiro, contendo ambos os genes alterados. A sobrevivência e a ação patogênica de SG não foi modificada pela alteração simples de um dos genes, mas tornou a cepa de SG completamente atenuada quando os dois foram modificados. A mortalidade provocada pela cepa selvagem de SG foi de 64,52%, enquanto que não observou-se mortalidade alguma no grupo de aves infectadas com SGNal r"cobs"cbiA. Estudos futuros deverão ser realizados para elucidar este processo fisiológico bacteriano e para avaliar a utilização desta cepa de SG como cepa vacinal.

Performance of a vaccine containing Anaplasma centrale and attenuated strains of Babesia bigemina e Babesia bovis against bovine babesioses and anaplasmosis / Avaliação do desempenho de uma vacina constituída por Anaplasma centrale e cepas atenuadas de Babesia bigemina e Babesia bovis contra a tristeza parasitária bovina

One hundred and thirty adult cattle imported from a tick-free region (Uruguay) were immunized in Brazil with a frozen trivalent vaccine containing Anaplasma centrale and attenuated Babesia bigemina and B. bovis. Thirty five days after the second dose of the vaccine, the animals ware challenged with virulent strains of the haemoparasites. At the day and 30 days after the challenge, it was taken serum samples from 28 animals, in order to evaluate the packed cell volume (PCV) and for serological tests. The results indicated that the animals resisted well the challenge with the virulent strains as showed by the absence of severe clinical symptoms during the observation period. The indirect ELISA test revealed serum antibodies against the three infeccious agents (B. bigemina - Overall Optical Density (OD) = 0.356, B. bovis = 0.320 and A. marginale - 0.295) in vaccineted animals, which raised significantly after challenging with virulent strains (B. bigemina - Overall OD = 0.664, B. bovis = 0.637and A. marginale = 0.549).

Cento e trinta novilhas importadas do Uruguai foram imunizadas no Brasil com uma vacina trivalente congelada, contendo Anaplasma centrale e cepas atenuadas de Babesia bigemina e B. bovis. Trinta e cinco dias após a vacinação, os animais foram desafiados com cepas virulentas de B. bigemina, B. bovis e A. marginale. No dia do desafio c 30 dias após, foram colhidas amostras de sangue do 26 animais para a avaliação de hematócrito e teste sorológico. Os animais resistiram bem ao desafio com as cepas virulentas, pela ausência de sinais clínicos da Tristeza Parasitária Bovina (TPB) durante o período de observação. O teste de ELISA indireto mostrou que os animais vacinados apresentavam anticorpos contra os três agentes da TPB (B. bigemina - Densidade Óptica (DO) média = 0,356, B. bovis = 0.320 e A. marginale = 0.295), aumentando significativamente após o desafio (B. bigemina - DO média = 0.664, B. bovis = 0.637 e A. marginale = 0.549).

A hematologic and electrophoretic study in puppies vaccinated against canine distemper virus and canine parvovirus / A hematologic and electrophoretic study in puppies vaccinated against canine distemper virus and canine parvovirus

Background: Canine distemper is a contagious multisystemic viral disease that affects canines and others carnivores. Canine parvovirus infection is one of the most important viral diseases in young dogs. Side effects of vaccine generally include fever, lethargy and local inflammation. Complementary exams are important to evaluate the strenght of immungenic stimulation. This study was aimed at evaluating hematological and electrophoretic alterations in puppies after inoculation of live attenuated vaccine against canine distemper virus and canine parvovirus.Materials, Methods & Results: Five non-breeding newborn dogs of the same litter were used. Animals received three subcutaneous injection of 1mL (at days 0, 21 and 42). Blood was collected at day 0 (day of vaccination) and for three times for each dose: at days 7, 14 and 21 (first dose); at days 28, 35 and 42 (second dose); and at days 49, 56 and 63 (third dose). Blood containing ethylenediaminetetraacetic acid as anticoagulant was used for hematological evaluation. The total serum protein were determined by the biuret method, using commercial reagent, according to fabricant instructions. Serum was used for protein fractionation by using cellulose acetate strip electrophoresis. A decrease in platelet count was observed at days 7 and 28 post-vaccination. Lymphocyte number increased 88.4%, as well as the level of the protein

Background: Canine distemper is a contagious multisystemic viral disease that affects canines and others carnivores. Canine parvovirus infection is one of the most important viral diseases in young dogs. Side effects of vaccine generally include fever, lethargy and local inflammation. Complementary exams are important to evaluate the strenght of immungenic stimulation. This study was aimed at evaluating hematological and electrophoretic alterations in puppies after inoculation of live attenuated vaccine against canine distemper virus and canine parvovirus.Materials, Methods & Results: Five non-breeding newborn dogs of the same litter were used. Animals received three subcutaneous injection of 1mL (at days 0, 21 and 42). Blood was collected at day 0 (day of vaccination) and for three times for each dose: at days 7, 14 and 21 (first dose); at days 28, 35 and 42 (second dose); and at days 49, 56 and 63 (third dose). Blood containing ethylenediaminetetraacetic acid as anticoagulant was used for hematological evaluation. The total serum protein were determined by the biuret method, using commercial reagent, according to fabricant instructions. Serum was used for protein fractionation by using cellulose acetate strip electrophoresis. A decrease in platelet count was observed at days 7 and 28 post-vaccination. Lymphocyte number increased 88.4%, as well as the level of the protein

Performance of a vaccine containing Anaplasma centrale and attenuated strains of Babesia bigemina e Babesia bovis against bovine babesioses and anaplasmosis. / Avaliação do desempenho de uma vacina constituída por Anaplasma centrale e cepas atenuadas de Babesia bigemina e Babesia bovis contra a tristeza parasitária bovina.

One hundred and thirty adult cattle imported from a tick-free region (Uruguay) were immunized in Brazil with a frozen trivalent vaccine containing Anaplasma centrale and attenuated Babesia bigemina and B. bovis. Thirty five days after the second dose of the vaccine, the animals ware challenged with virulent strains of the haemoparasites. At the day and 30 days after the challenge, it was taken serum samples from 28 animals, in order to evaluate the packed cell volume (PCV) and for serological tests. The results indicated that the animals resisted well the challenge with the virulent strains as showed by the absence of severe clinical symptoms during the observation period. The indirect ELISA test revealed serum antibodies against the three infeccious agents (B. bigemina - Overall Optical Density (OD) = 0.356, B. bovis = 0.320 and A. marginale - 0.295) in vaccineted animals, which raised significantly after challenging with virulent strains (B. bigemina - Overall OD = 0.664, B. bovis = 0.637and A. marginale = 0.549).  

Cento e trinta novilhas importadas do Uruguai foram imunizadas no Brasil com uma vacina trivalente congelada, contendo Anaplasma centrale e cepas atenuadas de Babesia bigemina e B. bovis. Trinta e cinco dias após a vacinação, os animais foram desafiados com cepas virulentas de B. bigemina, B. bovis e A. marginale. No dia do desafio c 30 dias após, foram colhidas amostras de sangue do 26 animais para a avaliação de hematócrito e teste sorológico. Os animais resistiram bem ao desafio com as cepas virulentas, pela ausência de sinais clínicos da Tristeza Parasitária Bovina (TPB) durante o período de observação. O teste de ELISA indireto mostrou que os animais vacinados apresentavam anticorpos contra os três agentes da TPB (B. bigemina - Densidade Óptica (DO) média = 0,356, B. bovis = 0.320 e A. marginale = 0.295), aumentando significativamente após o desafio (B. bigemina - DO média = 0.664, B. bovis = 0.637 e A. marginale = 0.549).