Benefit of Dietary Supplementation with Bacillus subtilis BYS2 on Growth Performance, Immune Response, and Disease Resistance of Broilers.

A strain of Bacillus subtilis (B. subtilis) BYS2 was previously isolated from Mount Tai, which is located in Tai'an City in the Shandong Province of China. The strain was then stored in the Environmental Microbiology Laboratory at Shandong Agricultural University. To evaluate the effect of the bacterium preparation in broiler production, we fed the bacterium (106 CFU/g) to 1-day-old broilers and continued this feeding for 6 weeks to analyze its effect on growth and immune performance. We found that the average weight of the bacterium-fed group increased by 17.19% at weeks 5 compared to the control group (P < 0.05). The height of the villi in the duodenum and jejunum and the ratio of villi to crypt were significantly increased in the bacterium-fed group at weeks 5 (P < 0.05). Also, the IgG in the serum of broilers in the experimental group increased by 31.60% (P < 0.05) and IgM 30.52% (P < 0.05) compared with those in the control group. The expressions of the major pattern recognition receptors (PRRs), antiviral proteins, pro-inflammatory cytokines, and ß-defensins were significantly higher than those in the control group (P < 0.05). Meanwhile, the bursa immune organ indices of broilers in the experimental group were significantly higher than those in the control group (P < 0.05). Also, after 5 weeks of continuous feeding, when infected with Escherichia coli (E. coli) O1K1 and Newcastle disease virus (NDV) F48E8, the content of bacteria and virus in tissues and organs of the experimental group decreased significantly, and the survival rate of infected chickens increased by 31.1% and 17.7%, respectively (P < 0.05). These results show that the anti-infective B. subtilis BYS2 could, to some extent, replace antibiotics to promote growth, improve innate immunity, and enhance disease resistance in broilers.

Immune Response of Salmonella Challenged Broiler Chickens Fed Diets Containing Gallipro®, a Bacillus subtilis Probiotic.

This study was conducted to investigate the effect of feeding a probiotic, Bacillus subtilis, on antibody titers against Newcastle and infectious bursal viruses in broiler chickens challenged with Salmonella enterica serotype Enteritidis. One hundred and sixty 1-day-old broiler chicks were randomly assigned to four treatments in a completely randomized design. The treatments were negative control, probiotic-treated group, challenged group, and challenged probiotic treated group. Salmonella challenging decreased (P < 0.05) the relative weights of spleen and bursa. Inclusion of probiotic to diet of challenged chickens increased the relative weight of spleen, but had no effect on the relative weight of bursa. There were no differences for the antibody titers of chickens between negative control and probiotic-treated group. Salmonella challenging decreased (P < 0.05) antibody titers against Newcastle and infectious bursal viruses. Improvements in the antibody titers were observed by the addition of probiotic to diet of these chickens. The results showed that dietary inclusion of probiotic had no significant effect on immune parameters of chickens at non-contaminated environment, display a greater efficacy at environment contaminated with pathogen and can improve immune responses of infected chickens.

[Effect difference between two segments in invariant chain CLIP on humoral immune].

OBJECTIVE: To compare the effect between two segments (PBS and GBS) of Class II -associated invariant chain peptide (CLIP) of invariant chain (Ii) on humoral immune by immune carrier. METHODS: First six hybrids containing Newcastle disease virus (NDV) epitope F2 and Ii segments (Cyt/TM/F2, Cyt/TM/F2/GBS, Cyt/TM/PBS/F2, Cyt/TM/F2/TRIM, Cyt/TM/F2/GBS/TRIM, Cyt/TM/PBS/F2/TRIM) were reconstructed respectively. Then they were inserted into the prokaryotic expression vector pET-32a and transformed into E. coli Rosetta (DE3) to induce the expression of the recombinant proteins. Finally mice were immunized with these purified fusion proteins, the specific antibody titers were detected with ELISA, to compare and analyze the effect among different groups on the immune response. RESULTS: All the six groups immunized with these hybrids increased antibody titers (from 1.5-fold to 4.9-fold, respectively) compared with the group immunized with F2 alone. Within the above six groups, the hybrids containing either PBS or GBS had higher antibody titers from 1.6-fold to 2.4-fold than the hybrids without the both segments. However, the group of the hybrid containing PBS had a 1.5-fold antibody titer higher than the group of GBS hybrid. CONCLUSION: Ii cytosolic and transmembrane domains could increase the immune response, while the segment PBS behaved better than GBS in an immune vector based on Ii.

NLRC5 deficiency does not influence cytokine induction by virus and bacteria infections.

Nucleotide-binding domain and leucine rich repeat containing gene family receptors (NLRs) are cytosolic proteins that respond to a variety of pathogen and host components to induce inflammatory cytokines. NLRC5 is a recently identified member of the NLR family that has been implicated in positive and negative regulation of antiviral innate immune responses. To clarify whether NLRC5 controls antiviral innate immunity in vivo, we generated NLRC5-deficient mice. Macrophages and dendritic cells derived from NLRC5-deficient mice induced relatively normal levels of IFN-ß, IL-6, and TNF-α after treatment with RNA viruses, DNA viruses, and bacteria. The serum cytokine levels after polyinosinic-polycytidylic acid infection were also comparable between control and NLRC5-deficient mice. NLRC5 overexpression promoted IL-1ß production via caspase-1, suggesting that NLRC5 constitutes an inflammasome. However, there was no reduction of IL-1ß in NLRC5-deficient cells in response to known inflammasome activators, suggesting that NLRC5 controls IL-1ß production through an unidentified pathway. These findings indicate that NLRC5 is dispensable for cytokine induction in virus and bacterial infections under physiologic conditions.

Combined exposure to cyanobacterial biomass, lead and the Newcastle virus enhances avian toxicity.

Under environmental conditions, wild birds can be exposed to multiple stressors including natural toxins, anthropogenic pollutants and infectious agents at the same time. This experimental study was successful in testing the hypothesis that adverse effects of cyanotoxins, heavy metals and a non-pathogenic immunological challenge combine to enhance avian toxicity. Mortality occurred in combined exposures to naturally occurring cyanobacterial biomass and lead shots, lead shots and Newcastle vaccination as well as in single lead shot exposure. Mostly acute effects around day 10 were observed. On day 30 of exposure, there were no differences in the liver accumulation of lead in single and combined exposure groups. Interestingly, liver microcystin levels were elevated in birds co-exposed to cyanobacterial biomass together with lead or lead and the Newcastle virus. Significant differences in body weights between all Pb-exposed and Pb-non-exposed birds were found on days 10 and 20. Single exposure to cyanobacterial biomass resulted in hepatic vacuolar dystrophy, whereas co-exposure with lead led to more severe granular dystrophy. Haematological changes were associated with lead exposure, in particular. Biochemical analysis revealed a decrease in glucose and an increase in lactate dehydrogenase in single and combined cyanobacterial and lead exposures, which also showed a decreased antibody response to vaccination. The combined exposure of experimental birds to sub-lethal doses of individual stressors is ecologically realistic. It brings together new pieces of knowledge on avian health. In light of this study, investigators of wild bird die-offs should be circumspect when evaluating findings of low concentrations of contaminants that would not result in mortality on a separate basis. As such it has implications for wildlife biologists, veterinarians and conservationists of avian biodiversity.

Isolamento e caracterização biológica da amostra JAP99 do vírus da doença de Newcastle isolada de patos domésticos (Neta sp) no Rio de Janeiro / Isolation and biological characterization of JAP99 Newcastle disease virus isolated from domestic ducks (Neta sp) in Rio de Janeiro State

O vírus da doença de Newcastle (VDN) é o agente etiológico de uma das doenças mais importantes da avicultura e tem sido identificado na maioria das espécies aviárias, silvestres e domésticas. Neste trabalho, obteve-se o isolamento em ovos embrionados da amostra denominada JAP99 do VDN, a partir de fezes de patos coletadas no Município de Japeri, RJ. A amostra foi identificada pela técnica da inibição da hemaglutinação (HI) e a caracterização biológica da patogenicidade do novo isolado foi realizada no Laboratório Regional Animal, MAPA, Campinas, SP (LARA-Campinas). Pela inoculação intracerebral em pintos de um dia (IPIC), o índice foi 1,4. Na inoculação intravenosa em aves de seis semanas (IPIV), o índice encontrado foi 0,0 (zero) e o tempo médio de morte embrionária (TMME) foi de 62 horas. Estes resultados indicaram que o isolado JAP99 é mesogênico para galinhas comerciais, oferecendo risco para a avicultura industrial.

Neurochemical and neuroendocrine responses to Newcastle disease virus administration in mice.

Mice injected intraperitoneally with Newcastle disease virus (NDV) responded with increased plasma concentrations of ACTH and corticosterone and increased hypothalamic concentrations of the tryptophan and of the norepinephrine catabolite, 3-methoxy,4-hydroxyphenylethyleneglycol (MHPG) and the serotonin catabolite, 5-hydroxyindoleacetic acid (5-HIAA). Two different strains of NDV, a lentogenic and a mesogenic one, elicited dose-dependent effects in these responses. Both strains elicited near maximal responses at doses around 1000 hemagglutination units. The maximal effects on ACTH, corticosterone and MHPG occurred around 2 h, but the effects on tryptophan and 5-HIAA were greatest at 8 h. Similar responses in plasma corticosterone, and cerebral tryptophan and 5-HIAA were observed following i.p. injection of polyinosinic-polycytidylic acid, but MHPG was not altered. The cyclo-oxygenase inhibitor, indomethacin, had little effect on the NDV-induced increases in plasma corticosterone and ACTH, and hypothalamic indolamines, but essentially ablated the MHPG response. The effect of NDV on plasma corticosterone, like that of endotoxin (LPS), was prevented by hypophysectomy, suggesting that the pituitary was required for these responses. These endocrine and neurochemical responses to NDV resemble those to interleukin-1 (IL-1) and LPS. Therefore we tested mice pretreated with the IL-1-receptor antagonist. This treatment prevented the neurochemical and plasma ACTH and corticosterone responses to IL-1, but did not alter those to LPS, and prevented the endocrine and neurochemical responses to NDV in approximately half of the animals. Thus IL-1 may be a mediator of the responses to NDV, but additional factors may also be involved.

Myxo and paramyxovirus surface structures: antigenic and functional studies

Amostras de vírus da Doença de Newcastle (cepas SO-93, B1 e La Sota) e da Influenza (cepa A/PR/8/34 (H1N1) foram analisadas especificamente quanto à presença de carboidratos específicos, de comprovada importância no processamento das atividades biológicas virais. As amostras virais foram concentradas por sedimentaçäo, avaliadas quanto ao seu teor proteico e título hemaglutinante e, entäo, estudadas quanto aos seus resíduos carboidratos. O estudo realizado com uso de lectinas e açúcar específico (inibidor de Concanavalina A) revelou a importância e especificidade do caráter carboidratado de estruturas peptídicas de superfície viral, o que pode ser observado através da inibiçäo do aparecimento das linhas de precipitaçäo entre Concanavalina A e soro específico para vírus da Doença de Newcastle

An oligonucleotide probe that distinguishes isolates of low virulence from the more pathogenic strains of Newcastle disease virus.

A synthesized oligonucleotide, termed cleavage probe (NDV-CL), has been designed to complement the cleavage-activation site of the fusion gene of the Texas GB isolate of Newcastle disease virus (NDV). This oligonucleotide probe, 21 bases in length, bound with RNA from velogenic strains of NDV tested in a slot-blot hybridization assay. The probe also recognized RNA from the mesogenic strains used in this assay, although no signal was observed with RNA isolated from lentogenic NDVs or with that from other common avian viruses used as controls. This probe did not recognize RNA from isolates of other paramyxovirus serotypes (PMV-2 or PMV-3) included in this study. The ability of this probe to distinguish lentogenic NDVs, which cause little or no clinical disease, from those strains that may produce severe morbidity and/or mortality suggests a potential use for the probe in a molecular diagnostic assay.

Immunohistochemical detection of Newcastle disease virus in chickens.

An immunoperoxidase histochemical technique utilizing a monoclonal primary antibody was developed for detection of Newcastle disease virus (NDV) antigen in tissues from chickens. The technique was applied to trachea, lung, spleen, Harderian gland, and cecal tonsil harvested from specific-pathogen-free (SPF) chickens at 2, 5, 7, 10, and 14 days postinoculation (PI) with NDV, and to corresponding tissues from commercial broiler chickens representing 30 cases of spontaneous respiratory disease. Positive staining occurred in the cytoplasm of respiratory epithelial cells in the trachea or bronchi of NDV-inoculated SPF chickens at 5 and 7 days PI. Staining also occurred in the respiratory epithelium of the trachea and bronchi of commercial broilers from seven of 30 cases of spontaneous respiratory disease. These results indicate that the immunoperoxidase technique has value as a rapid diagnostic test for Newcastle disease.

Occurrence of velogenic viscerotropic Newcastle disease in pet and exotic birds in 1991.

In 1991, velogenic viscerotropic Newcastle disease (VVND) was diagnosed in domestic psittacine birds in six states: Illinois, Indiana, Michigan, Texas, California, and Nevada. In the first four states, the disease assumed outbreak proportions. The affected psittacine birds--yellow-headed Amazon parrots (Amazona ochrocephala oratrix), yellow-naped Amazon parrots (Amazona ochrocephala auropalliata), cockatiels (Nymphicus hollandicus), and conures (unknown species)--exhibited respiratory and/or central nervous system signs. The velogenic viscerotropic Newcastle disease virus (VVNDV) was isolated from cloacal and tracheal swabs and various tissues, such as the lung, trachea, distal intestine, and spleen. The origin of the birds could not be established. The disease in the six states was promptly controlled, with no evidence that domestic poultry had been exposed. Also, VVNDV was isolated from quarantined birds intended for importation into the United States. Included were 53 moustached parakeets (Psittacula alexandri fasciata), a mynah (Gracula religiosa), a drongo (Dicrurus sp.), and three partridges (family Phasianidae). Groups of birds that yielded VVNDV were denied entry into the United States. Birds that are illegally imported and therefore not tested for the presence of foreign animal pathogens are a potential source of VVNDV and a threat to domestic poultry and caged birds.

[The tenacity of Newcastle disease virus (LaSota) in the excrement of laying hens in different housing systems]. / Tenazität von Newcastle-Disease-Virus (LaSota) in den Exkrementen von Legehennen in unterschiedlichen Haltungssystemen.

The survival time of NDV (LaSota) in the excrement of layers was in summer (winter) 22 and 18 days (26 and 36 days) in two cage houses, 14 and 18 days (36 and 33 days) in two floor-pen houses, as well as 8 days (54 and 68 days) in two dropping store places. By one week staying in battery cages and following the storage in dropping store place after 47 or 50 days NDV (LaSota) could not be reisolated. Besides environment factors, temperature, pH and dry matter was the thermic effect very significant. The comparison of the tenacity of NDV (LaSota) in different housing systems was besides of the quantitative determination of survival time supported through the application of a life time distribution test.

Isolation and characterisation of Newcastle disease virus strain in a feral dove (Stigmatopelia senegalensis) in Nigeria.

An isolate of Newcastle disease virus (NDV) was obtained from a feral dove, (Stigmatopelia senegalensis). The isolate was shown to have a mean-death-time of 96 h and an intracerebral pathogenicity index of 0.10. It was immunogenic but not pathogenic for 6-week old chicks on experimental infection. Based on these observations the isolate was classified as a lentogenic strain. The role of such isolates of NDV from wild birds on the Nigerian poultry industry is discussed.

Characterisation of Newcastle disease viruses isolated in India.

Eleven Newcastle disease virus (NDV) isolates obtained from outbreaks of disease in chickens (9) and Japanese quail (2) in Tamil Nadu, India were characterised in pathogenicity tests, antigenically, using mouse monoclonal antibodies (MAbs), and other established tests devised to distinguish between different strains. All 11 isolates were shown to be highly virulent for chickens. In indirect immunoperoxidase tests used to assess the ability of a panel of 28 MAbs to bind to infected cell cultures, 10 of the isolates showed an identical reaction pattern, the other isolate (No. 4) failed to react with one MAb which bound to cells infected with the other isolates. Isolates 9 was unstable at pH 3 while the other 10 were stable. All other properties were shared by the 11 isolates.

Avian paramyxovirus serotype 1 (Newcastle disease virus)--infections in falcons.

From eight falcons and one pigeon which died from NDV over a period of 15 months in Dubai, United Arab Emirates, PMV-1 viruses were isolated on quail embryo cell cultures. The identification of all 9 strains were achieved with the haemagglutination inhibition test against polyclonal chicken PMV-1 antiserum, against mouse monoclonal antibodies as well as with the immunoperoxidase test. Intracerebral pathogenicity index and intravenous pathogenicity index tests were also carried out. Although the virus isolates in this study fell into two distinct groups, the overall clinical symptoms displayed by the falcons tailed to demonstrate any trends or specificity unique to a group. The isolate obtained from a pigeon was similar to the isolates from one group of the falcons and showed no identity with the pigeon variant virus.

Pathogenicity of two strains of Newcastle disease virus in the grey-breasted helmet guinea fowl.

Thirty-five 6-week-old guinea fowl keets, seronegative for maternal antibodies to Newcastle disease virus, were infected with Herts strain (33/56) and Kumarov strain of Newcastle disease virus intramucularly (IM) or intranasally (IN). Clinical signs were first noticed four days post infection (PI) in the group infected IM but five days PI in the group infected IN with Herts strain of Newcastle disease virus. These clinical signs were similar in both groups and included anorexia, droopiness, huddling together, greenish diarrhoea and marked cachexia. Prominent nervous signs, including spasms of the head and neck, were observed in groups infected with Herts strain. The major gross lesions observed were emaciation with prominent keel bone, empty intestinal tract and distended gall bladder in most keets. The histological lesions were characterised by meningoencephalitis, necrosis and loss of lymphocytes from splenic and lymphoid aggregates. There was muscular degeneration and necrosis in the gizzard and mild pulmonary congestion and oedema in some keets. Neither gross or microscopic lesions were observed in keets that had received the Kumarov strain.

Characterisation of an antigenically unusual virus responsible for two outbreaks of Newcastle disease in the Republic of Ireland in 1990.

Antigenic characterisation of two highly virulent virus isolates from outbreaks of Newcastle disease on two closely connected farms in County Monaghan, Republic of Ireland, in 1990 showed the viruses to be indistinguishable but unlike other Newcastle disease viruses so far tested. However, they appeared to be antigenically closest to avirulent viruses isolated from waterfowl from several countries and from chickens in Northern Ireland in 1986. Despite the antigenic differences, chickens vaccinated with a live commercial Hitchner B1 vaccine were protected against intramuscular challenge with one of the virulent isolates.

[Newcastle disease in Algeria. Study of the pathogenic properties of Newcastle disease virus strains isolated in Algeria]. / La maladie de New-castle en Algérie. Etude des propriétés pathogéniques des souches virales de la maladie de New-castle isolées en Algérie.

The Newcastle disease in Algeria: Study of pathogenic properties of the Newcastle disease viral strains isolated in Algeria. The study of pathogenic characters of 14 strains of Newcastle disease virus isolated from 1972 to 1982 was carried out: The following testS were realized: the mean lethal time for the chicken embryo, the pathogenic power index by intravenous way, the pathogenic power index by intracerebral way, the hemagglutination spectrum and the thermal stability of the hemagglutination. In short we can say that these isolated 14 strains are fast-growing.

A concentraçäo de interferon humano de membranas amnióticas: II - métodos químicos / Concentrations of the human amniotic membrane interferon: II - Chemical methods

Foi investigada a concentraçäo do interferon humano de membranas amnióticas (IFN-MA) por precipitaçäo através de métodos químicos. O IFN-MA foi produzido em âmnios infectados com o vírus da doença de Newcastle em meio contendo diferentes concentraçöes de soro. A atividade anti-vírica das preparaçöes foi titulada em células Vero infectadas com o vírus da encefalomiocardite de camundongo ou o vírus Sindbis. A precipitaçäo de IFN-MA, consideradas baixas. Empregando-se sulfato de âmnio em preparaçöes com 1//de soro, pode-se recuperar quantidades acima de 60//. O método mais adequado de concentraçäo de IFN-MA contendo 1//de soro foi a precipitaçäo com ácido tricloroactico e ressuspenso do matrial com tampäo clorto pH2, com recuperaçöes em torno de 100//da atividade. As tentativas de concentrar IFN-MA preparado sem soro por estes métodos näo permitiu uma precipitaçäo adequada

Purification of human anmiotic membrane interferon by affinity chromatography

A purificaçäo do interferon humano de membranas amnióticas foi realizada para se obter preparaçöes purificadas para sua caracterizaçäo físico-química, estudo de suas atividades biológicas e ensaios terapêuticos. O interferon foi produzido em doença de Newcastle ou o vírus Sendai. A determinaçäo da atividade anti-vírica foi feita em células Vero infectadas com o vírus da encefalomiocardite de camundongo. As preparaçöes de interferon foram concentradas por precipitaçäo por ácido tricloroactivo e cromatografadas em "Blue-Sepharose- concanavalina A, Sepharose ligada ao dipeptídeo triptofnio (trf) - triptofânio ao dipeptídeo triptofânio-tirosina (tyr). Com a "Blue-Sepharose", o pH ótimo de ligaçäo do interferon foi 5,4. A aplicaçäo em batelada mostrou que foram ligadas 37.000 unidades de interferon por ml de gel, sendo portanto, mais eficiente que a aplicaçäo em coluna, na qual somente foram ligadas 17.750 unidades por ml de gel. As fraçöes do pico de atividade foram purificadas 274 vezes, com 12//de recuperaçäo do interferon em um experimento, com um total de 78//de recuperaçäo e um fator de purificaçäo atingido foi de apenas 6,7; os ligantes Sepharose trp-trp e trp-tyr mostraram uma ligaçäo baixa do interferon, menor que 10.000 unidades por ml de gel