Identification of Avibacterium paragallinarum serovar B-1 from severe infectious coryza outbreaks in Panama

The isolation and identification of Avibacterium paragallinarum serovar B-1 from severe infectious coryza outbreaks in broiler breeders in Panama is reported for the first time. Infectious coryza was absent for over a decade in the breeder farms area. Disease outbreaks were characterized by an up to 45% drop in egg production and increased mortality. Use of a commercial trivalent bacterin and a strengthened biosecurity program prevented outbreaks in susceptible flocks in the farm.     

Evaluation of the protection conferred by a disinfectant against clinical disease caused by Avibacterium paragallinarum serovars A, B, and C from Argentina

This work evaluates the efficiency of the administration of the disinfectant N-alkyl dimethyl benzyl ammonium chloride (TIMSEN) in the prevention of the horizontal transmission of serovars A, B, and C of Avibacterium paragallinarum, the causative agent of avian infectious coryza. This disinfectant was administered in drinking water (50 ppm) and once or twice per day by coarse spray (800 ppm, 8 ml per m3 during 3 seconds). In three trials conducted with vaccinated birds, the disinfectant reduced the clinical signs of infectious coryza significantly (P < 0.05). There was no significant effect when the product was used in a fourth trial with unvaccinated birds. Furthermore, the application of only one daily environmental spraying was sufficient to significantly reduce clinical signs. According to these results, in order to diminish the clinical signs of infectious coryza in birds vaccinated against A. paragallinarum, it is recommended to administer this disinfectant in drinking water and by environmental spraying.     

Study on the efficacy and safety of different antigens and oil formulations of infectious coryza vaccines containing an NAD-independent strain of Avibacterium paragallinarum

The present study was designed to assess and compare three different formulations of the new Onderstepoort infectious coryza (IC) quadrivalent vaccine, which contain an NAD-independent strain of Avibacterium paragallinarum (previously known as Haemophilus paragallinarum), and a commercial IC vaccine, not containing an NAD-independent strain, for their safety and ability to protect chickens of varying ages against virulent challenges with four different serovars of A. paragallinarum, including the NAD-independent strain of the C-3 serovar. Four groups of 140 chickens each were vaccinated at the age of 17 weeks and revaccinated at the age of 19 weeks with each of the four vaccine formulations. A similar sized group of non-vaccinated chickens was used as control. Two rounds of challenge were conducted: a group of chicken in each vaccination group was challenged between 31 and 35 weeks of age, while another group was challenged between 51 and 55 weeks of age. The "in-contact" challenge model was used in this experiment. For each vaccination group, the four challenge strains representing four local serovars were used in each challenge round. The efficacy of the vaccines was compared based on overall protection levels obtained and the duration of protection. The safety of the different vaccines was determined by the severity of post-vaccination reactions. The need for the incorporation of the NAD-independent strain in the vaccine was evidenced by the low protection level against NAD-independent challenge recorded in the group of birds vaccinated with the commercial vaccine. The results obtained confirmed not only the variation in virulence of different South African serovars, with serovar C-3 being the most virulent and serovar B having almost no virulence but also the age related increase in susceptibility. The importance of a suitable formulation of the vaccine is discussed.     

Isolation of serovar C-3 Haemophilus paragallinarum from Zimbabwe: A further indication of the need for the production of vaccines against infectious coryza containing local isolates of H. paragallinarum

Various isolates of Haemophilus paragallinarum, collected from a severe outbreak of infectious coryza in poultry from Zimbabwe, were serotyped and were found to belong to serovar C-3. Previously, isolates were serotyped using polyclonal antiserum produced against serogroup reference strains (0083 for serogroup A, 0222 for serogroup B and Modesto, or H-18 for serogroup C) of H. paragallinarum. In this case, polyclonal antiserum produced against these reference isolates were used, as well as polyclonal antiserum that has been raised specifically against the serovar C-3 isolate 46 C-3. When using the latter serum at a 1 in 50 dilution, no cross-reaction with other members of serogroup C were found. The severity of the disease outbreak in Zimbabwe, the vaccination history of the infected flocks on the sites and the isolation of the uniquely southern African serovar C-3, further highlights the need for vaccines composed of local isolates to control infectious coryza in regions where vaccination failures occur.     

Efficacy of a commercially available coryza vaccine against challenge with recent South African NAD-independent isolates of Haemophilus paragallinarum in chickens

In South Africa the incidence of NAD-independent Haemnophilus paragallinarum isolation from clinical cases is increasing. This study was carried out to test whether a commercially available coryza vaccine (Nobilis Coryza, Intervet International BV) could protect chickens against challenge with recent NAD-independent isolates. SPF chickens were vaccinated twice at 3 and 7 weeks of age and were challenged at 9 weeks of age with 5 different NAD-independent isolates of serotype A or C-3. The results after challenge show that the coryza vaccine induces good protection against challenge with the different South African NAD-independent isolates of H. paragallinarum, including serotype C-3.     

A cross-protection experiment in pigs vaccinated with Haemophilus parasuis serovars 2 and 5 bacterins, and evaluation of a bivalent vaccine under laboratory and field conditions.

Cross-protection between Haemophilus parasuis serovars 2 and 5 was examined in pigs using a bacterin based vaccine, and subsequently the safety and efficacy of a bivalent vaccine were evaluated. Upon intratracheal challenge of a serovar 2 or 5 strain, pigs immunized with a monovalent vaccine were protected against challenge with a homologous serovar strain, but not with a heterologous serovar strain. Immunization with a bivalent vaccine containing both serovars 2 and 5 bacterins conferred protection in pigs against lethal challenge with each of the serovar strains. A total of 86 pigs from two SPF herds were injected with the bivalent vaccine intramuscularly twice at a four-week interval. No adverse reactions following the vaccination were observed. On day 7 after the second vaccination, vaccinated and non-vaccinated control pigs from herd A were transferred to herd B, where Glasser's disease had broken out. Pigs in the control group developed clinical signs of the disease, and 6 of 8 (75%) pigs died until slaughter, in contrast with only 4 of 46 (9%) pigs in the vaccinated group. In herd C, where there was no outbreak of Glasser's disease, complement fixation antibody titer was raised only in the vaccinated group. A challenge experiment on days 20 and 79 after the second vaccination showed that only the vaccinated pigs were protected. From these findings, the safety and efficacy of the bivalent vaccine were confirmed under laboratory and field conditions.     

Preparation and use of an autogenous bacterin against infectious coryza in chickens

An experiment was conducted to determine the efficacy of an autogenous bacterin against infectious coryza from a local strain of Haemophilus paragallinarum in Morocco compared with a commercial vaccine. Hens were vaccinated with a single dose or two doses of the bacterin at 15 and 18 weeks of age. Both the autogenous and the commercial vaccine conferred significant protection against experimental challenge (94% and 88%, respectively). A single dose was less protective with both vaccines.     

Preparation and use of autogenous vaccine from Avibacterium paragallinarum (strain Tan 1-05) in layer chickens

The study was conducted to prepare and evaluate the use of autogenous vaccine from Avibacterium paragallinarum (strain Tan 1-05) in layer chickens. The results showed that all chickens vaccinated with autogenous vaccine with 10⁸ CFU/mL in aluminum phosphate gel developed MAT antibodies (GMT of 2.8 log₂ to 5.3 log₂) against A. paragallinarum infection. Moreover, the results indicated that all chickens (n = 6) selected from vaccinated chickens were protected against A. paragallinarum infection after challenge. No A. paragallinarum was isolated from these chickens. Nevertheless, all unvaccinated chickens did not develop antibodies, and all selected unvaccinated chickens (n = 6) showed clinical signs consistent with infectious coryza (IC) where two of them died from the disease after challenge. The findings from the present and previous studies showed that the development of an inactivated IC vaccine from local strains if optimized and adopted may be the best possible way of controlling this economically important poultry disease.     

Comparative efficacy of several disinfectants in suspension and carrier tests against Haemophilus parasuis serovars 1 and 5

The comparative efficacy of 16 active compounds (including the most commonly used chemical groups) and 10 commercial formulations against Haemophilus parasuis serovars 1 and 5 was studied. These organisms were tested in suspension and carrier tests in the presence and absence of serum as representative of organic matter. Chloramine-T and half of the formulations from commercial sources (most of them including quaternary ammonium compounds) were effective in both in vitro tests, regardless of the presence or absence of organic load. All 26 disinfectants except for an iodophor (0.1% available iodine) resulted in at least 3-log₁₀ reduction in colony-forming units in suspension test, and most of them resulted in the maximal level of detection (>6-log₁₀ reduction). On the other hand, disinfectants were not as effective in carrier test as in suspension test, and the presence of serum considerably reduced the activities of most of the compounds tested, especially in carrier test. These results suggest the importance of selecting suitable disinfection for routine use on surfaces contaminated with H. parasuis, particularly when organic matter is present. Chloramine-T and formulations 2 and 7–10 are recommended for a complete inactivation of H. parasuis in swine herds.     

Protection conferred by vaccination with Blacksburg and Komarov strains of Newcastle disease virus against Newcastle disease in Bangladesh

Summary An evaluation was undertaken of the efficacy of vaccination of day-old chicks with the Blacksburg (B1) strain of Newcastle disease virus (NDV) followed at various times by vaccination with the Komarov (K) strain. Antibody was detected by the haemagglutination inhibition (HAI) test one week after vaccination with B1 and titres peaked at three weeks and had declined to undetectable levels by nine weeks.After subsequent vaccination with K strain at five, seven or eight weeks of age levels of HAI antibody (titre 80 to 640) were detected after three weeks. Birds vaccinated at seven weeks were tested for antibody and resistance to challenge beyond 19 weeks of age. In this group the HAI titres remained constant (80 to 640) up to 32 weeks of age and then steadily declined to 10 to 20 at 44 weeks of age.A linear relationship between HAI titre and virus neutralising index (VNI) was demonstrated with a range of selected sera. Only birds with an HAI titre of 80 or greater resisted artificial challenge. It is recommended that, following B1 vaccincation at day-old and K vaccination at seven weeks old, revaccination with K strain should be performed at intervals of not more than seven months.

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Proteccion Conferida Mediante Vacunacion Con Las Cepas Blacksburg Komarov Del Virus De Newcastle En Bangladesh

Resumen Se llevó a cabo una evaluación de la eficacia de la vacunación de pollitos de un día, con la cepa Blacksburg (B1) del virus de la enfermedad de Newcastle, seguida de varias aplicaciones vacunales periódicas con la cepa Komarov (K). Se detectaron anticuerpos mediante la prueba de inhibición de la hemaglutinación, una semana después de la vacunación con B1, los títulos alcanzando el pico a las tres semanas y declinaron a niveles no detectables a las nueve semanas.Después de aplicaciones vacunales periódicas con la cepa K, a las cinco, siete u ocho semanas de edad, se detectaron anticuerpos mediante la prueba de inhibición de la hemaglutinación (títulos 80 a 640), después de tres semanas. Las aves vacunadas a las siete semanas, recibieron una descarga y se les hizo prueba de anticuerpos, después de las 19 semanas de edad. En este grupo los títulos de anticuerpos permanecieron constantes (80 to 640) hasta las 32 semanas de edad y después declinaron lentamente a 10 ó 20 a las 44 semanas de edad.Se demostró una relación lineal entre los títulos hallados y el índice viral neutralizante, en un rango de sueros seleccionados. Solamente las aves con títulos de 80 o más altos resistieron la descarga artificial. Se recomienda que, seguidamente a la vacunación con B1 a un día de edad y con K a las siete semanas, las revacunaciones con la cepa K debería hacerse con intervalos de no más de siete meses.

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Protection Conferee Contre La Maladie De Newcastle Au Bangladesh Par La Vaccination A L'Aide Des Souches Blacksburg Et Komarov De Virus De La Maladie De Newcastle

Résumé Une evaluation de l'efficacité de la vaccination de poussins d'un jour à l'aide des souches Blackburg (B1) du virus de la maladie de Newcastle, suivie à différents moments par la vaccination à l'aide de la souche Komarov (K), a été entreprise. Les anticorps furent détéctés par inhibition de l'hemagglutination (HAI), une semaine après la vaccination avec la souche B1 et le titre atteignit un pic à 3 semaines pour décliner jusqu'à des niveaux indétectables à neuf semaines. A la suite de vaccinations consécutives avec la souche K à cinq, sept ou huit semaines, les niveaux d'anticorps HAI furent détectés après trois semaines. Les oiseaux vaccinés à sept semaines furent examinés pour les anticorps et la résistance à une épreuve au delà de 19 semaines d'âge. Dans ce groupe les titres HAI sont restés constants (80 à 640) jusqu'à l'àge 32 semaines, puis ont diminué régulièrement jusqu'à 10 à 20 à l'âge de 44 semaines.Une relation linéaire entre le titre HAI et l'index de neutralisation du virus (VNI) a été démontrée sur une gamme de sérums choisis. Seuls les oiseaux ayant un titre HAI de 80 ou plus ont résisté à l'épreuve artificielle. On recommande que, après une vaccination B1 à un jour, et une vaccination K à l'âge de sept semaines, une revaccination K soit réalisée à des intervalles n'excédant pas sept mois.

     

Development and application of an enzyme-linked immunosorbent assay to detect antibodies against prevalent Salmonella serovars in swine in southern Brazil.

The implementation of Salmonella control programs in the pork production chain demands rapid and cost-effective methods to assess the prevalence of infection in pig herds. The objective of the present study was to develop an in-house enzyme-linked immunosorbent assay (ELISA) based on S. Typhimurium lipopolysaccharides (LPS) to measure the prevalence of infection caused by Salmonella in swine herds. Coating antigen was produced by phenol extraction of S. Typhimurium culture. After standardization of ELISA test conditions, the assay was validated by testing serum samples on different animal categories: pigs orally inoculated with S. Typhimurium and sentinel animals in contact with them, naturally infected animals, colostrum-deprived piglets, and bacterin-immunized pigs. Seroconversion was observed in inoculated pigs (7 days postinfection [DPI]) and in the sentinels (21 DPI). Nonspecific reactions were not detected in the sera of colostrum-deprived animals. Serum samples from animals immunized with Salmonella Agona, Salmonella Derby, Salmonella Panama, and Salmonella Bredeney bacterins showed marked cross-reaction with the LPS from the serovar Typhimurium. Moreover, positive results obtained with the in-house ELISA were associated with Salmonella isolation in 75 infected pig herds. Comparisons with 2 commercial kits showed a linear correlation coefficient of 0.847 between the in-house ELISA and kit A and 0.922 with kit B but a low agreement in the qualitative results. In conclusion, the newly developed in-house ELISA based on S. Typhimurium LPS can be a useful tool to determine the intensity of Salmonella sp. infection in swine herds.     

A laboratory study of an inactivated bivalent iron restricted Salmonella enterica serovars Enteritidis and Typhimurium dual vaccine against Typhimurium challenge in chickens

A commercial inactivated iron restricted Salmonella Typhimurium and Salmonella Enteritidis vaccine was used to vaccinate chicks at 1 day and again at 4 weeks of age, with challenge by a high and a low dose of S. Typhimurium given either orally or by contact with seeder birds inoculated orally with a high dose of S. Typhimurium. In all three challenge regimes, the shedding of challenge strain was reduced significantly (p < 0.05) in vaccinated birds compared with unvaccinated controls. Vaccination reduced colonisation of internal organs after challenge by contact seeder birds. However, no effect of vaccination upon colonisation of internal organs after either high or low oral challenge was apparent. In conclusion, the data indicate that the vaccine should be a useful tool in the control of S. Typhimurium infection in chickens.     

Role of polymorphonuclear neutrophils (PMNs) and NK cells in the protection conferred by different vaccines against Chlamydophila abortus infection

Ovine enzootic abortion (OEA) is caused by Chlamydophila abortus, an intracellular bacterium which acts by infecting the placenta, causing abortion in the last term of gestation. The main prevention strategy against OEA is the vaccination of flocks. An effective vaccine against C. abortus must induce a Th1-like specific immune response, which is characterized by the early production of IFN-gamma and the activation of CD8(+)T cells. Moreover, vaccine effectiveness could be modulated by the functioning of the innate immunity. The purpose of this study was to ascertain how polymorphonuclear neutrophils (PMNs) and NK cells might influence vaccine-induced protection. The live attenuated 1B vaccine and two inactivated experimental vaccines, adjuvated with aluminium hydroxide (AH) or QS-21 (QS), were used in PMN-depleted or NK cell-depleted mice. For PMN depletion, RB6-8C5 monoclonal antibody, which recognizes GR1(+) receptors (Robben, P.M., LaRegina, M., Kuziel, W.A., Sibley, L.D. 2005. Recruitment of Gr-1(+) monocytes is essential for control of acute toxoplasmosis. The Journal of Experimental Medicine 201, 1761-1769.) was used, while for NK cell-depletion the anti-asialo GM1 polyclonal antibody was used. The depletion of PMNs caused 100% mortality in non-vaccinated mice (NV) and 60% mortality in the AH-vaccinated mice by day 10 p.i., while both groups showed a significant increase in their bacterial burden in the liver by day 4 p.i. The depletion of NK cells caused mortality only in the NV group (50% by day 10 p.i.), although this group and the 1B vaccinated mice showed an increased bacterial burden in the liver at day 4 p.i. Our results suggest that the importance of PMNs in inactivated vaccines depends on the adjuvant chosen. The results also demonstrated that the importance of NK cells is greater in live vaccines than in inactivated vaccines.     

Production and characterization of monoclonal antibodies against Actinobacillus (Haemophilus) pleuropneumoniae serotype 2

Four hybridoma cell lines producing monoclonal antibodies (MAbs) against Actinobacillus (Haemophilus) pleuropneumoniae were established by fusion of mouse myeloma and spleen cells obtained from mice immunized with a serotype 2, strain SH-15. Enzyme-linked immunosorbent assay-inhibition tests with antigens obtained from 12 serotype strains of A. pleuropneumoniae and 9 other gram-negative bacteria showed that all the MAbs bound to only serotype 2 strains of A. pleuropneumoniae. The epitopes recognized by the MAbs were located on a carbohydrate moiety of lipopolysaccharide (LPS) of the organism, which was sensitive to periodate oxidation. In immunoblotting analyses of LPS obtained from A. pleuropneumoniae serotype 2, all the four MAbs reacted specifically with the characteristic ladder bands of LPS detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. These results suggest that O-antigen side chains of the LPS are one of the antigenic determinants responsible for the serotype-specificity of A. pleuropneumoniae.     

Protection of mice against the lethal effect of an intraperitoneal infection with Haemophilus (Actinobacillus) pleuropneumoniae after vaccination with capsular proteins

Haemophilus (Actinobacillus) pleuropneumoniae Serotypes 5 and 7 capsular antigens (CA-1) were precipitated from culture supernatants with N-cetyl-N,N,N,-trimethylammonium bromide (Cetavlon). CA-1 contained a carbohydrate to protein ratio of 2:1 for Serotype 5 and 3:1 for Serotype 7. Glucosamine and uronic acid were detected in CA-1 from both serotypes suggesting that the capsule contained hyaluronic acid. All mice immunized intraperitoneally with CA-1 vaccine were protected from death when challenged with 10X the LD50 of the homologous but not the heterologous serotype. Oil adjuvants and the use of young (6 h) cultures were necessary for CA-1 vaccines to be protective. Deproteinization of CA-1 with chloroform and butanol followed by pronase treatment resulted in failure to protect mice from death. The protective capsular protein antigen in CA-1 vaccine may not originate from the outer membrane (OM) since repeated washing of the OM to elute the capsular protein antigen rendered the OM vaccine completely nonprotective for mice. Vaccines prepared from cell-wall lipopolysaccharide also were nonprotective for mice. Passive immunization of mice with anti-CA-1 antibody produced in rabbits to Serotype 5 was highly protective (P less than 0.01) for mice when challenged with 10X the LD50 of the homologous serotype.