Development and Licensing of Avian Vaccines - Perspective from the Vaccine Industry.

Veterinary vaccines need to be authorized by relevant authorities before they can be used in the field. This paper briefly describes the development and authorization process of vaccines. It also highlights important regulatory trends, challenges and opportunities from the veterinary vaccine industry standpoint in EU, US, Asia and Latin America, with a specific focus on avian vaccines' relevant topics.

Desarrollo y concesión de licencias para vacunas aviares: Perspectiva de la industria de producción de vacunas. Las vacunas veterinarias deben ser autorizadas por las autoridades pertinentes antes de que puedan usarse en el campo. Este artículo describe brevemente el proceso de desarrollo y autorización de vacunas. También destaca importantes tendencias regulatorias, desafíos y oportunidades desde el punto de vista de la industria de vacunas veterinarias en la Unión Europea, los Estados Unidos, Asia y América Latina, con un enfoque específico en los temas relevantes de las vacunas aviares.

Direct transverse load profile determination using the polarization-dependent loss spectral response of a chirped fiber Bragg grating.

The determination of stress profiles created by transverse loads was proved to be important in different domains, such as structural health monitoring and biomechanics, and, more specifically, in the prostheses domain. In this paper, we report an original method to estimate the transverse load profile from the polarization-dependent loss (PDL) spectrum of a chirped fiber Bragg grating (CFBG). This method makes use of the relationship between the integration of the PDL of a CFBG, and the force profile has the advantage of not requiring any iterative method to estimate the transverse load profile. The relationship linking the integration of the PDL and the force profile is demonstrated using an analytical approximation of the transmission spectrum of CFBGs. The validity of this method for the determination of non-uniform load profiles is then shown using a numerical analysis. An experimental demonstration is finally reported using a 48 mm-long CFBG subject to different step transverse load profiles.

Distribution profiling of a transverse load using the DGD spectrum of chirped FBGs.

In this paper, we propose a new method to determine the longitudinal distribution of a non-uniform transverse force applied to an optical fiber. For that purpose, we use a chirped fiber Bragg grating (CFBG) for which we monitor the polarization parameters in reflection. In particular, we demonstrate that the differential group delay (DGD) spectrum of the CFBG is an imprint of the load profile so that it can be used for the shape determination of an applied load. Thereafter, we discuss the influence of the CFBG parameters on the achievable accuracy and resolution of our technique. An experimental validation is finally reported with two 48 mm long CFBGs subject to step transverse load profiles.

Influence of the optical fiber type on the performances of fiber-optics current sensor dedicated to plasma current measurement in ITER.

In this paper, we compare, by means of simulations using the Jones formalism, the performances of several optical fiber types (low birefringence and spun fibers) for the measurement of plasma current in international thermonuclear experimental reactor (ITER). The main results presented in this paper concern the minimum value of the ratio between the beat length and the spun period, which allows meeting the ITER current measurement specifications. Assuming a high-birefringence spun fiber with a beat length of 3 mm, we demonstrate that the minimum ratio between the beat length and the spun period is 4.4 when considering a 28 m long sensing fiber surrounding the vacuum vessel. This minimum ratio rises to 10.14 when a 100 m long lead fiber connecting the interrogating system to the sensing fiber is taken into account.

Simulation of vibration-induced effect on plasma current measurement using a fiber optic current sensor.

An accurate measurement of the plasma current is of paramount importance for controlling the plasma magnetic equilibrium in tokamaks. Fiber optic current sensor (FOCS) technology is expected to be implemented to perform this task in ITER. However, during ITER operation, the vessel and the sensing fiber will be subject to vibrations and thus to time-dependent parasitic birefringence, which may significantly compromise the FOCS performance. In this paper we investigate the effects of vibrations on the plasma current measurement accuracy under ITER-relevant conditions. The simulation results show that in the case of a FOCS reflection scheme including a spun fiber and a Faraday mirror, the error induced by the vibrations is acceptable regarding the ITER current diagnostics requirements.

Evaluation of immunogenicity and protective efficacy of a Microsporum canis metalloprotease subunit vaccine in guinea pigs.

In order to identify protective immunogens against Microsporum canis infection, a purified recombinant keratinolytic metalloprotease (r-MEP3) was tested as a subunit vaccine in experimentally infected guinea pigs. Both humoral and cellular specific immune responses developing towards r-MEP3 were evaluated, by enzyme-linked immunosorbent assay and by in vitro lymphocyte transformation tests respectively. Vaccination induced a strong antibody response, and a significant but transient lymphoproliferative response against the protein. However, the protocol failed to prevent fungal invasion or development of dermatophytic lesions. These results show that under the present experimental conditions, r-MEP3 specific antibodies are not protective against a challenge exposure. They also suggest that in the same model, the induction of cell-mediated immunity towards r-MEP3 is not sufficient, indicating the need for further research in the field of specific immune mechanisms involved in M. canis dermatophytosis.

A recombinant 31.5 kDa keratinase and a crude exo-antigen from Microsporum canis fail to protect against a homologous experimental infection in guinea pigs.

A Microsporum canis recombinant 31.5 kDa keratinase and a M. canis crude exo-antigen were tested as vaccines in an experimental infection model in guinea pigs. Animals were vaccinated subcutaneously three times at two-week intervals with either the keratinase, the exo-antigen or the adjuvant alone. Cutaneous challenge was performed blindly. Both humoral and cellular-specific immune responses to M. canis antigens were evaluated every 14 days, while a blind evaluation of clinical lesion development and fungal persistency in skin were monitored weekly. Vaccination induced very high and significant (P < 0.01) antibody responses towards both antigens. High cell-mediated immune responses to both immunogens were also induced by vaccination. After challenge, however, scores reflecting the severity of dermatophytic lesions did not differ significantly between vaccinated and control groups at any time after challenge. These results suggest that, in the guinea pig, the induction of specific immune responses against the M. canis-secreted antigens used in this study are not protective against challenge exposure.

Recombinant expression and antigenic properties of a 31.5-kDa keratinolytic subtilisin-like serine protease from Microsporum canis.

A secreted 31.5-kDa keratinolytic subtilase (SUB3; AJ431180) is thought to be a Microsporum canis virulence factor and represents a candidate for vaccination trials. In this study, the recombinant keratinase (r-SUB3) was produced by the Pichia pastoris expression system and purified to homogeneity. Recombinant SUB3 displayed identical biochemical properties with the native protease. Experimentally cutaneously infected guinea pigs showed specific lymphoproliferative response towards r-SUB3, while no specific humoral immune response was induced except for one animal. The heterologous expression of SUB3 provides a valuable tool for addressing further investigations on the role of this keratinase in the specific cellular immune response and on its use in vaccination trials in the cat.

Humoral and cellular immune response to a Microsporum canis recombinant keratinolytic metalloprotease (r-MEP3) in experimentally infected guinea pigs.

In order to better understand the host-fungus relationship in Microsporum canis dermatophytosis and to identify major fungal antigens, the immune response to a crude exoantigen preparation and to a purified recombinant keratinolytic metalloprotease (r-MEP3) was evaluated in guinea pigs experimentally infected with M. canis. Humoral and cellular immune responses were assessed from day 0 to day 57 post-infection (PI), the former by enzyme-linked immunosorbent assay (ELISA) and the latter via a lymphocyte proliferation assay. Infected guinea pigs developed humoral and cellular responses to both M. canis exoantigen and r-MEP3, while no specific immune response to these antigens was observed in control animals. This is the first report on the development of both humoral and cell-mediated immune responses to a purified keratinase in M. canis dermatophytosis.

Isolation of a Microsporum canis gene family encoding three subtilisin-like proteases expressed in vivo.

Microsporum canis is the main agent of dermatophytosis in dogs and cats and is responsible for frequent zoonosis. The pathogenesis of the disease remains largely unknown, however. Among potential fungal virulence factors are secreted keratinolytic proteases, whose molecular characterization would be an important step towards the understanding of dermatophytic infection pathogenesis. M. canis secretes a 31.5 kDa keratinolytic subtilisin-like protease as the major component in a culture medium containing cat keratin as the sole nitrogen source. Using a probe corresponding to a gene's internal fragment, which was obtained by polymerase chain reaction, the entire gene encoding this protease named SUB3 was cloned from a M. canislambdaEMBL3 genomic library. Two closely related genes, termed SUB1 and SUB2, were also cloned from the library using as a probe the gene coding for Aspergillus fumigatus 33 kDa alkaline protease (ALP). Deduced amino acid sequence analysis revealed that SUB1, SUB2, and SUB3 are secreted proteases and show large regions of identity between themselves and with subtilisin-like proteases of other filamentous fungi. Interest ingly, mRNA of SUB1, SUB2, and SUB3 were detected by reverse transcriptase nested-polymerase chain reaction from hair of experimentally infected guinea pigs. These results show that SUB1, SUB2, and SUB3 encode a family of subtilisin-like proteases and strongly suggest that these proteases are produced by M. canis during the invasion of keratinized structures. This is the first report describing the isolation of a gene family encoding potential virulence-related factors in dermatophytes.

Secreted metalloprotease gene family of Microsporum canis.

Keratinolytic proteases secreted by dermatophytes are likely to be virulence-related factors. Microsporum canis, the main agent of dermatophytosis in dogs and cats, causes a zoonosis that is frequently reported. Using Aspergillus fumigatus metalloprotease genomic sequence (MEP) as a probe, three genes (MEP1, MEP2, and MEP3) were isolated from an M. canis genomic library. They presented a quite-high percentage of identity with both A. fumigatus MEP and Aspergillus oryzae neutral protease I genes. At the amino acid level, they all contained an HEXXH consensus sequence, confirming that these M. canis genes (MEP genes) encode a zinc-containing metalloprotease gene family. Furthermore, MEP3 was found to be the gene encoding a previously isolated M. canis 43.5-kDa keratinolytic metalloprotease, and was successfully expressed as an active recombinant enzyme in Pichia pastoris. Reverse transcriptase nested PCR performed on total RNA extracted from the hair of M. canis-infected guinea pigs showed that at least MEP2 and MEP3 are produced during the infection process. This is the first report describing the isolation of a gene family encoding potential virulence-related factors in dermatophytes.