Vertebral Osteomyelitis in Broilers: A Review

ABSTRACT Locomotor diseases are still a challenge in modern poultry. Vertebral osteomyelitis (VO) is an emerging disease in broilers worldwide. The inflammatory process in the affected thoracic vertebra (T4) and subsequent spinal cord compression leads to clinical signs related to locomotor impairment, inadequate feeding and drinking, and increased mortality in the affected flocks. The pathogenesis of the disease is poorly understood and Enterococcus cecorum is the bacterium most frequently associated with the disease. However, other bacteria such as E. faecalis, E. durans, Escherichia coli and Staphylococcus aureus have been recently detected in cases of the disease, raising questions regarding its etiopathogenesis. As many questions about VO in broilers remain unanswered, knowledge on its prevention, control and treatment is limited. In this review, we compile and discuss the currently available information concerning VO in broilers and highlight important aspects of the disease.

Occurrence and Epidemiology of Fannia Spp. (Diptera: Fanniidae) in Laying Poultry Farms in State of Minas Gerais, Brazil

ABSTRACT Aspects related to epidemiology of flies belonging to Fannia genus were studied in order to determine predisposing factors for its occurrence in layer hens houses of commercial farms in the state of Minas Gerais, Brazil. An observational, analytical and sectional survey was conducted to verify major epidemiological aspects associated to infestations. This research was accomplished starting from an adapted secondary database of and structured with information on the presence and absence of ectoparasites and arthropods, characteristics of chicken houses, the poultry, type of facilities, geographical location and the management. The study was conducted in four hundred and thirty one houses located in forty three farms visited in 2012 year in the state of Minas Gerais. Ten farms (23.25%) and sixty eight houses (15,77%) were observed the presence diptera belonging to Fannia genus. The specie Fannia admirabilis was found in four farms (9,30%), Fannia albitarsis in two farms (4,65%) and only in one farm (2.32%) was observed the presence of Fannia canicularis. For three farms 6,98% was no possible to identify the species only Fannia genus. The epidemiological aspects for the occurrence of these flies were investigated using Poisson regression models. Characteristics such as farm, city of farm, presence of Histeridae beetles, type of facility (cage, on the floor with or without bedding) and the existence of mechanical removal of manure were associated with the occurrence of Fannia spp.

Detecting reproductive system abnormalities of broiler breeder roosters at different ages.

The objective of this study was to detect the reasons of rooster's fertility decrease at 50 weeks of age. Therefore, the reproductive system of broiler breeder roosters was laparoscopic, macroscopic and histopathology evaluated, and a comparison of the anatomical aspect with the sperm analysis and birds' age was realized. Cobb roosters (n = 59) were distributed into two groups (30 and 50 weeks). Evaluations were performed with laparoscopy, macroscopy and histopathology, and seminal quality, blood serum testosterone concentration and weight were also determined. The old roosters presented smaller testicle size, higher intensity epididymal lithiasis and lower testicle sperm production, compared to the young roosters. The use of the endoscope could easily distinguish a normal-sized testicle than an atrophic one. Four old roosters with severe testicular atrophy did not show spermatogenesis, although three still had sperm in the ejaculate. This would falsely indicate a wrong diagnosis of normal fertility before the testicular atrophy took place. In conclusion, in addition to the weight increase with age, the testicular atrophy and impairment of sperm production seemed to be the main reason to the decrease in the rooster's fertility at 50 weeks of age. Therefore, the use of the laparoscopy as a way to detect the roosters with testicular atrophy before 50 weeks of age and their removal from them flock could be useful as a diagnostic tool to prevent the birds' fertility loss.

Histomorphological changes by epididymal lithiasis in roosters

Epididymal lithiasis (EL) histopathology is described using light and electronic microscopy in roosters (Gallus gallus domesticus) naturally affected by EL in Minas Gerais, Brazil. The histologic and morphological changes by EL in roosters was performed regarding cellular and subcellular details through light and electron microscopy. Efferent ductules epithelium lysosomal increase in size and numbers, membrane rupture, cellular vacuolation, ciliary loss, basal membrane degeneration, inflammatory reaction with mononuclear infiltrations, edema, epithelial and vascular endothelium losses were described. All industrial and freerange chickens showed EL in varying degrees in the efferent ductules (ED). However, ED altered areas did not correlate with the presence of luminal stones. Non-ciliated ED epithelium cells presented several atypically large lysosomes. Plicae loss and basal vacuoles were observed in the epithelium of dilated regions. Cellular cilia loss and apical cytoplasmic membrane rupture resulted in leakage of the cytoplasmic contents to the ED lumen, and ED epithelium desquamation occurred with or without lesion to the basal membrane. Basal membrane alterations were associated with profound sub-epithelial connective tissue damage. Aggregations of desquamated epithelium and spermatozoa were seen in the lumen of ED and compact aggregates were considered the basis for calculi formation. The widespread occurrence and high severity of EL lesions are indicative of the importance of EL as a cause of infertility in male chickens.

Histomorphological changes by epididymal lithiasis in roosters

Epididymal lithiasis (EL) histopathology is described using light and electronic microscopy in roosters (Gallus gallus domesticus) naturally affected by EL in Minas Gerais, Brazil. The histologic and morphological changes by EL in roosters was performed regarding cellular and subcellular details through light and electron microscopy. Efferent ductules epithelium lysosomal increase in size and numbers, membrane rupture, cellular vacuolation, ciliary loss, basal membrane degeneration, inflammatory reaction with mononuclear infiltrations, edema, epithelial and vascular endothelium losses were described. All industrial and freerange chickens showed EL in varying degrees in the efferent ductules (ED). However, ED altered areas did not correlate with the presence of luminal stones. Non-ciliated ED epithelium cells presented several atypically large lysosomes. Plicae loss and basal vacuoles were observed in the epithelium of dilated regions. Cellular cilia loss and apical cytoplasmic membrane rupture resulted in leakage of the cytoplasmic contents to the ED lumen, and ED epithelium desquamation occurred with or without lesion to the basal membrane. Basal membrane alterations were associated with profound sub-epithelial connective tissue damage. Aggregations of desquamated epithelium and spermatozoa were seen in the lumen of ED and compact aggregates were considered the basis for calculi formation. The widespread occurrence and high severity of EL lesions are indicative of the importance of EL as a cause of infertility in male chickens.

Detection of Mycoplasma gallisepticum in dead captive psittacines in Belo Horizonte, Brazil

Mycoplasma gallisepticum (Mg) infection of wild native Brazilian psittacines (Psittaciformes) which died of any cause during sorting, rehabilitation, or conservation, was investigated by PCR. Two previously described PCR methodologies using Mg specific primers were employed for the analyses of 140 swab samples (cloaca, trachea, or palatine cleft). Average positive Mg detection in cloacal swabs was 51.9%, with 80.0% (n=5) of Blue-and-yellow Macaws (Ara ararauna), 60.0% (n=3) Dusky Parrots (Pionus fuscus), 52.5% (n=59) Amazon Parrots (Amazona aestiva), 50.0% (n=2) Orange-winged Parrots (Amazona amazonica), 50.0% (n=2) Jandaya Parakeetsor Jandaya Conures (Aratinga jandaya), 0% (n=2) Golden Conures or Golden Parakeets (Guarouba guarouba), and 0% (n=2) Hyacinth Macaws (Anodorhynchus hyacinthinus). Palatine cleft swab sampling was more sensitive to detect Mg, with 85.4% (n=17) detection rate, as compared to 67.4% (n=46) obtained with tracheal samples, and 53.5% (n=77) with cloacal swabs. The surprisingly high Mg incidence in psittacines kept in conservation or triage environments is possibly due to the proximity or cohabitation with several bird species during confinement and housing psittacines of different origins together. The implementation of biosecurity measures and species-specific facilities is recommended.

Lead poisoning mortality in wild passeriformes and its detection in free-range chicken eggs in Southern Minas Gerais, Brazil

The mortality of passerines native of Southern Minas Gerais, Brazil, in August 2006 and the examination of sentinel free-range chicken eggs in August 2007 in an area surrounding a car battery lead smelter plant is described. The high levels of lead, as detected in the passerines (4.80-12.74 mg/kg) at the onset of mortality and at the follow-up sampling of the free-range chicken eggs (25.02-35.21 mg/kg in shells, 0.41-1.36 mg/kg in yolks and 0.40-0.75 mg/kg in the albumins), used as environmental sentinels, indicated the continuing lead pollution.

Lead poisoning mortality in wild passeriformes and its detection in free-range chicken eggs in Southern Minas Gerais, Brazil

The mortality of passerines native of Southern Minas Gerais, Brazil, in August 2006 and the examination of sentinel free-range chicken eggs in August 2007 in an area surrounding a car battery lead smelter plant is described. The high levels of lead, as detected in the passerines (4.80-12.74 mg/kg) at the onset of mortality and at the follow-up sampling of the free-range chicken eggs (25.02-35.21 mg/kg in shells, 0.41-1.36 mg/kg in yolks and 0.40-0.75 mg/kg in the albumins), used as environmental sentinels, indicated the continuing lead pollution.

Detection of Mycoplasma gallisepticum in dead captive psittacines in Belo Horizonte, Brazil

Mycoplasma gallisepticum (Mg) infection of wild native Brazilian psittacines (Psittaciformes) which died of any cause during sorting, rehabilitation, or conservation, was investigated by PCR. Two previously described PCR methodologies using Mg specific primers were employed for the analyses of 140 swab samples (cloaca, trachea, or palatine cleft). Average positive Mg detection in cloacal swabs was 51.9%, with 80.0% (n=5) of Blue-and-yellow Macaws (Ara ararauna), 60.0% (n=3) Dusky Parrots (Pionus fuscus), 52.5% (n=59) Amazon Parrots (Amazona aestiva), 50.0% (n=2) Orange-winged Parrots (Amazona amazonica), 50.0% (n=2) Jandaya Parakeetsor Jandaya Conures (Aratinga jandaya), 0% (n=2) Golden Conures or Golden Parakeets (Guarouba guarouba), and 0% (n=2) Hyacinth Macaws (Anodorhynchus hyacinthinus). Palatine cleft swab sampling was more sensitive to detect Mg, with 85.4% (n=17) detection rate, as compared to 67.4% (n=46) obtained with tracheal samples, and 53.5% (n=77) with cloacal swabs. The surprisingly high Mg incidence in psittacines kept in conservation or triage environments is possibly due to the proximity or cohabitation with several bird species during confinement and housing psittacines of different origins together. The implementation of biosecurity measures and species-specific facilities is recommended.

Epididymal lithiasis in Gallus gallus domesticus

A study on the occurrence of epididymal lithiasis (EL) in Gallus gallus domesticus of different breeds and from different geographical regions of Brazil was carried out. Forty breeding roosters were collected in the states Santa Catarina, São Paulo, Minas Gerais, Espírito Santo and Goiás, representing two specific pathogen free Leghorn lines (SPF), one commercial Leghorn layer breeder (Hy-Line) line, two commercial broiler breeder lines (Ross and Cobb), and two backyard chicken flocks. In commercial poultry breeders, EL was observed mainly after 55 weeks of age, and lithus resulted in lower fertility caused by inadequate maturation of spermatozoa and testicular atrophy, which evolved to more severe testicular dysfunction at 100 weeks of age. One hundred percent of the sampled roosters of the evaluated SPF lines presented epididymal lithiasis, which was also observed in all commercial genetic lines. The analysis of EL occurrence revealed no differences among regions and breeds: all regions and genetic lines presented epididymal calculi and the occurrence seemed higher in the more intensively-reared chickens. As for the backyard roosters, 50% presented calculi. Considering the occurrence of EL in the SPF flocks, its primary association with infectious bronchitis virus (IBV) infection was discarded, despite its contribution to fertility loss.

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.

Occurrence of chicken anemia virus in backyard chickens of the metropolitan region of Belo Horizonte, Minas Gerais

The occurrence of CAV in backyard chickens in the metropolitan area of Belo Horizonte, Brazil, was evaluated. The spleen and thymus of chickens from different origins were collected for DNA extraction and nested-PCR. CAV genome was detected in 30% of the flocks (n=20) examined. CAV origin for backyard chickens is speculated, taking into consideration its widespread incidence in the chicken industry, the contamination of live vaccines with CAV prior to its eradication from SPF flocks, and the use of attenuated CAV vaccines.

Occurrence of chicken anemia virus in backyard chickens of the metropolitan region of Belo Horizonte, Minas Gerais

The occurrence of CAV in backyard chickens in the metropolitan area of Belo Horizonte, Brazil, was evaluated. The spleen and thymus of chickens from different origins were collected for DNA extraction and nested-PCR. CAV genome was detected in 30% of the flocks (n=20) examined. CAV origin for backyard chickens is speculated, taking into consideration its widespread incidence in the chicken industry, the contamination of live vaccines with CAV prior to its eradication from SPF flocks, and the use of attenuated CAV vaccines.

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.

Epididymal lithiasis in Gallus gallus domesticus

A study on the occurrence of epididymal lithiasis (EL) in Gallus gallus domesticus of different breeds and from different geographical regions of Brazil was carried out. Forty breeding roosters were collected in the states Santa Catarina, São Paulo, Minas Gerais, Espírito Santo and Goiás, representing two specific pathogen free Leghorn lines (SPF), one commercial Leghorn layer breeder (Hy-Line) line, two commercial broiler breeder lines (Ross and Cobb), and two backyard chicken flocks. In commercial poultry breeders, EL was observed mainly after 55 weeks of age, and lithus resulted in lower fertility caused by inadequate maturation of spermatozoa and testicular atrophy, which evolved to more severe testicular dysfunction at 100 weeks of age. One hundred percent of the sampled roosters of the evaluated SPF lines presented epididymal lithiasis, which was also observed in all commercial genetic lines. The analysis of EL occurrence revealed no differences among regions and breeds: all regions and genetic lines presented epididymal calculi and the occurrence seemed higher in the more intensively-reared chickens. As for the backyard roosters, 50% presented calculi. Considering the occurrence of EL in the SPF flocks, its primary association with infectious bronchitis virus (IBV) infection was discarded, despite its contribution to fertility loss.

Antimicrobial susceptibility of Clostridium perfringens strains isolated from broiler chickens

Clostridium perfringens is a normal inhabitant of the intestinal tract of chickens as well as a potential pathogen that causes necrotic enteritis and colangio hepatitis. The minimum inhibitory concentration (MIC) of seven different compounds used for therapy, growth promotion or prevention of coccidiosis was determined by agar dilution method for 55 C. perfringens strains isolated from the intestines of broiler chickens. All strains showed high susceptibility to penicillin, avilamycin, monensin and narasin. Only 7.3% of the strains showed an intermediated sensitivity to lincomycin, and 49 (89.1%) were considered susceptible. For tetracycline and bacitracin, 41.8% and 47.3% of strains, respectively, were considered resistant.

Clostridium perfringens é um habitante normal da microbiota intestinal de frangos, sendo um agente potencialmente patogênico, causador de enterite necrótica e colangio-hepatite. A concentração inibitória mínima (CIM) de sete drogas utilizadas na terapêutica, como agentes promotores de crescimento ou na prevenção de coccidiose foi determinada pelo método de diluição em ágar para 55 estirpes de C. perfringens isoladas do intestino de frangos de corte. Todas as estirpes revelaram alta sensibilidade à penicilina, avilamicina, narasin e monensina, apenas 7,3% demonstraram CIM intermediário para lincomicina e 89.1% foram consideradas sensíveis. Para tetraciclina e bacitracina, 41,8% e 47.3% das amostras, respectivamente, foram consideradas resistentes.

Distribuição de anticorpos para herpesvírus bovino 1 em rebanhos bovinos

The distribution of neutralizing antibodies to bovine herpesvirus 1 (BHV 1) was studied in 21 beef and dairy cattle herds, in four different age groups. The results were analyzed and related with answers given to a questionnaire proposed to the farmers. The frequency rates of neutralizing antibodies for the BHV 1 were compared according to the dairy or beef herds and age of the animals. The animal husbandry and age influenced the distribution of neutralizing antibodies to this virus.

A distribuição de anticorpos neutralizantes para o herpesvirus bovino 1 (HVB 1) foi estudada em quatro faixas etárias de bovinos, em 21 rebanhos de leite e de corte. Os resultados da sorologia foram analisados e relacionados com as respostas de questionários aplicados aos responsáveis pelos rebanhos. As taxas de freqüência de anticorpos neutralizantes para o HVB 1 foram comparadas segundo a aptidão e a faixa etária. Fatores como tipo de manejo e idade dos animais influenciaram na distribuição de anticorpos para o HVB 1.

Estudo comparativo da virulência de amostras de vacina do vírus da doença de Newcastle em galinhas SPF por meio da análise morfométrica da espessura traqueal

Virulence of three vaccinal lentogenic strains (La Sota, Ulster and VG-GA) of the Newcastle disease virus (NDV) was determined by morphometric analysis of tracheal thickness, in 45-day-old SPF chickens (n=12), free from NDV antibodies. Tracheal thickness was evaluated 1, 2, 3, 4, 6, and 8 days after intratracheal vaccination. La Sota strain induced higher tracheal swelling than the others, during most of the time of the experiment. Maximum swelling of tracheal mucosa occurred at the thirdday after vaccination. At that time, La Sota and Ulster had the same virulence, and both caused higher swelling of tracheal mucosa than VG-GA strain.

A virulência das amostras de vacinas lentogênicas La Sota, Ulster e VG-GA do vírus da doença de Newcastle (VDN) foi determinada por análise morfométrica da espessura da traquéia de galinhas (n=12) de 45 dias de idade, livres de anticorpos anti-VDN, vacinadas por via intra-traqueal. As traquéias foram avaliadas 1, 2, 3, 4, 6 e 8 dias pós-vacinação. A amostra La Sota induziu maior espessamento da traquéia no decorrer de todo período experimental. Ao terceiro dia pós-vacinação, período em que as traquéias se apresentaram mais espessas, as amostras La Sota e Ulster não diferiram em sua virulência, sendo ambas mais virulentas do que a amostra VG-GA, induzindo maior espessamento de traquéia e causando lesões histológicas mais severas.

Influenza Aviária: Uma Revisão dos Últimos Dez Anos

Avian influenza is an exotic disease in the Brazilian poultry. The National Avian Health Surveillance Program (PNSA) maintains permanent monitoring of AVES of all domestic species, including imported genetic material for the poultry industry, for example chickens (Gallus gallus formadomestica), turkeys (Meleagris gallopavo formadomestica), quail (Coturnix coturnix japonica), ducks (Anas), elite, grand grandparent and grandparent stocks for layers and broilers, as well as species more recently exploited for production, for example, the exotic ostriches (Struthio camelus) or native rheas (Rhea americana). The breeding stocks are monitored by regular sampling for serology, virology and bacteriology, principally looking for Newcastle disease, influenza, salmonellas and mycoplasmas, as established by the PNSA, in addition to monitoring vaccination responses to, for example, infectious bursal disease and infectious bronchitis. The PNSA establishes the rules for the control and eradication of Newcastle disease and avian influenza (Projeto de Vigilância..., 2001), including the need for differential diagnosis, according to the following major actions: I. Notification of outbreaks (and laboratory confirmation at the LARA-Campinas); II. Sanitary assistance to the outbreaks; III. Disinfection and sanitation measures; IV. Sanitary slaughter; V. Sanitary depopulation; VI. Vaccination of flocks and emergency strategies; VII. Control and monitoring of susceptible flocks; VIII. Other sanitary measures; The active surveillance and outbreak attention policies require the diagnosis and differential diagnosis of ND and AI, following the code described by OIE and the PNSA, as follows: 1- Zoning, interdiction and sampling for laboratory confirmation; 2- Registry of AVES, including species, category and numbers; applicable to a 10 km radius: restriction to transportation of animals and materials possible source of infection and propagation; disinfection of sites of entry and exit to affected properties; epidemiological surveillance; 3- Laboratory confirmation by isolating and characterizing AIV: hemagglutinating agent isolated in SPF eggs not inhibited by NDV specific serum, characterized as AIV by detecting antigens of the nucleoprotein and/or matrix and subtyped by assaying for hemagglutinin and neuraminidase subtypes (immunodiffusion, enzyme immunoassays or molecular methodology); 4- Slaughter and cremation of all avian individuals, residues, meats and eggs of all affected and neighboring properties (3 km radius), cleaning and disinfection of premises; sanitary depopulation for 21 days (minimum). 5- Allow transportation for slaughter or incubation within the vigilance area (10 km radius). 6- Prohibit fairs, markets, expositions etc., within the vigilance area (10 km radius). 7- Apply these measures for a minimum of 21 days after disinfection (which follows slaughter, cremation and cleaning) and prohibit transportation of animals and residues/products of properties within the protection area (3 km radius) and for 15 days within the vigilance area (10 km radius). The certification of HPAIV area is according to OIE and PNSA regulations and considers Brazil as a free country, applicable as follows: 1- HPAIV is not diagnosed by the active surveillance for the last 3 years; 2- 6 months after an outbreak of HPAIV is diagnosed, birds and residues/products are destroyed. The major species (chickens and turkeys) maintained in the Brazilian poultry industry employ the state-of-the-art production technology. Flocks are managed with the forefront knowledge in biosecurity, housing, permanent evaluation of critical points and quality and vaccination programs, which guarantee the prevention of most health problems. The geographic localization of the Brazilian poultry industry may also play a role in the absence of influenza outbreaks, in addition to the lower rate of replication of AIV in migratory birds during their subtropical season. Migratory routes may also concentrate in areas not occupied by poultry industry. Added to that, the biosecurity/housing and quality system of the industry may represent the step further to prevent health problems caused by transmissible infectious diseases such as influenza, considering that, for instance, all chicken and turkey industrial flocks are kept in houses, in contrast to open field farming, especially for ducks, geese and turkeys, kept on migratory routes of countries facing influenza problems.

A influenza aviária é doença exótica no Brasil. O sistema de vigilância implementado pelo Programa Nacional de Sanidade Avícola (PNSA) mantém monitoração permanente das aves das principais espécies domésticas, tanto do material genético importado para a indústria avícola, por exemplo, da espécie das galinhas (Gallus gallus formadomestica), perus (Meleagris gallopavo formadomestica), codornas (Coturnix coturnix japonica), patos (Anas), primários (elite), bisavós e avós para postura ou corte, como aves de espécies de exploração mais recente, exóticas, por exemplo avestruzes (Struthio camelus) ou nativas, por exemplo emas (Rhea americana). Os plantéis de reprodutores em produção são também acompanhados por amostragens periódicas, conforme previsto no PNSA, além da monitoração das respostas aos programas de vacinação, por exemplo, contra bronquite infecciosa e doença infecciosa bursal. O PNSA estabelece as normas de atuação para o controle e erradicação da doença de Newcastle (ND) e Influenza Aviária (AI) (Projeto de Vigilância, 2001), a saber: I - Notificação de focos da doença (e confirmação laboratorial no LARA-Campinas); II - Assistência a focos; III - Medidas de desinfecção; IV - Sacrifício sanitário; V - Vazio sanitário; VI - Vacinação dos plantéis ou esquemas emergenciais; VII - Controle e fiscalização dos animais susceptíveis; VIII - Outras medidas sanitárias; A vigilância e atenção ao foco exige o diagnóstico laboratorial e diferencial de AI e ND, que segue as normas do PNSA, conforme o sumário abaixo: 1- Interdição e coleta de materiais para exame laboratorial oficial; 2- Registro das aves: espécie(s), categoria(s), número(s), manutenção de aves; utensílios e produtos no local; proibição de trânsito de e para a(s) propriedade(s) em um raio de 10 km; controle de todos os animais e materiais possíveis fontes de propagação; desinfecção de vias de entradas e saídas à(s) propriedade(s); inquérito epidemiológico. 3- Confirmação laboratorial: isolamento de agente letal hemaglutinante em ovos embrionados de galinhas SPF, não inibido (inibição da hemaglutinação) ou não neutralizado (soroneutralização) por soro específico para o vírus da doença de Newcastle; caracterização do agente como vírus da influenza aviária (AIV) por detecção de antígenos da nucleoproteína e/ou matriciais de AIV e de seu subtipo por ensaios específicos para a caracterização da hemaglutinina e neuraminidase (imunodifusão, imunoenzimáticos ou moleculares). 4- Abate e destruição imediata (cremação) de todas as aves, resíduos, carnes e ovos da(s) propriedade(s) atingida(s) e vizinhas (raio de 3 km); limpeza e desinfecção das instalações; vazio sanitário (mínimo 21 dias); 5- Permitir o transporte para o abate ou incubação dentro da zona de vigilância (raio de 10 km). 6- Proibir feiras, exposições, mercados na zona de vigilância (10 km). 7- Aplicar estas medidas por mínimo de 21 dias após a destruição das fontes de propagação e desinfecção das instalações, proibir a retirada de aves e produtos na zona de proteção (3 km) por 21 dias e 15 dias na zona de vigilância (10 km). A certificação de área livre segue as normas da OIE e PNSA, considerando AI exótica no Brasil (país livre), e exige: 1- AI de alta patogenicidade não diagnosticada pelo sistema de vigilância pelos últimos 3 anos; 2- Um período de 6 meses após o abate, destruição das aves e resíduos e desinfecção após surto; O sistema de criação da avicultura predominante no Brasil (galinhas e perus) emprega a mais atual tecnologia e conhecimento científico na produção, no qual os plantéis são gerenciados com biossegurança, avaliação permanente dos pontos críticos, sistema de qualidade total e programas de vacinações que garantem a prevenção de inúmeros problemas sanitários. A prevenção de influenza aviária é especialmente favorecida por essas características. O sistema e tipo de construção (galpões) para o alojamento dos plantéis dessas espécies dificultam também o desafio eventualmente imposto pelas aves de vida livre. A localização geográfica da avicultura nacional, localizada fora das rotas migratórias das aves-reservatório, pode também exercer papel importante na ausência de focos de influenza no Brasil. Além disso, o baixo índice de replicação dos AIV nas aves migratórias durante a estada na região subtropical também influi para a menor ocorrência. As espécies de aves domésticas de importância comercial mais sensíveis à infecção e potencialmente envolvidas no papel de fonte de infecção, conforme citadas na literatura internacional, perus e patos, são mantidas em galpões industriais com sistema de biossegurança e de distribuição geográfica bastante restrita, em contraste com as criações dos países com relatos permanentes de surtos, em que se associam as condições de desafio naturais geográficas ditadas pelas rotas migratórias, mais alta replicação na ave na estação (países temperados) e a criação em campo aberto.

Production of monoclonal antibodies against conserved components of infectious bronchitis virus

Murine hybridomas producing IgG1 monoclonal antibodies (Mabs) against N and S2 proteins (53KDa and 82KDa, respectively) from avian infection bronchitis virus (IBV) strain M41 were generated by the fusion of a myeloma cell line (Sp2/0-Ag14) with spleen cells from Balb/c mice previously immunized with whole virus IBV M41. Post-fusion screening criterion was by ELISA and 36 positive hybrids were generated after fusions. Two hybrids specific to N (N3F10) and S2 (S12B2) proteins from M41 (serotype Massachusetts) were selected by western blotting. These Mabs recognized the Ark-99 (serotype Arkansas) and A5968 (serotype Connecticut) IBV strains in addition to M41. By ELISA, the Mab against the S2 (S12B2) recognized all reference and Brazilian strains (M41, SE-17, H52, 297, 283, PM-1, PM-2, PM-3, 351, 29-78 E 327) studied, while the Mab against N recognized only six (M41, SE-17, H52, 283, 327 e 297) strains. The Mab against S2 may become a useful tool for IBV detection on the routine diagnosis of infectious bronchitis, especially for helping the differential diagnosis of clinically and pathologically confusing diseases, while the Mab against N (N3F10) recognized a probably less conserved region among the strains and may be interesting to comparing IBV isolates.

Foram produzidos anticorpos monoclonais (AcM) da subclasse IgG1 contra as proteínas N (53KDa) e S2 (82KDa) do vírus da bronquite infecciosa das galinhas (VBIG) amostra M41. Os híbridos secretores originaram-se da fusão entre células de mielomas da linhagem Sp2/0-Ag14 e linfócitos B de camundongos Balb/c previamente imunizados com o vírus completo. O primeiro critério de seleção foi por ELISA, no qual 36 híbridos originados de duas fusões reagiram positivamente; destes, foram selecionados dois AcM que reagiram contra as proteínas N (N3D4) e S2 (S12B2) do VBIG da amostra M41 (sorotipo Massachusetts) no western blotting. Os mesmos AcM foram também capazes de reconhecer as estirpes Ark-99 (sorotipo Arkansas) e A5968 (sorotipo Connecticut) do VBIG no western blotting. No ELISA, o AcM contra S2 (S12b2) reconheceu 11 estirpes das 11 estudadas (M41, SE-17, H52, 297, 283, PM-1, PM-2, PM-3, 351, 29-78 E 327), enquanto o AcM contra a proteína N reconheceu apenas seis estirpes (M41, SE-17, H52, 283, 327 e 297). O anticorpo monoclonal contra S2 comportou-se como uma boa ferramenta de diagnóstico do VBIG, independentemente do sorotipo que pode ser aplicado em ensaios de rotina laboratorial, especialmente no diagnóstico das doenças que se confundem com a BIG nas galinhas, enquanto o AcM contra N (N3F10) demonstrou reconhecer uma região menos conservada entre as estirpes de VBIG e pode ser útil em estudos comparativos entre isolados de VBIG.