RESUMO Quarenta e nove ovos embrionados de Psittaciformes com embriões que morreram durante a incubação foram examinados, provenientes de criatório comercial de aves de espécies exóticas ou nativas, ou provenientes de instituição de conservação de espécies da avifauna. Os ovos foram classificados, de acordo com o momento da morte, em mortalidade precoce, intermediária ou tardia. Conforme a idade embrionária, embriões inteiros ou tecidos embrionários foram coletados para extração de DNA e cultivo bacteriológico em ágar contendo acetato de tálio, soro equino e penicilina. Entre os embriões de espécies exóticas, 37,5% (12/32) foram detectados positivos para Mycoplasma spp. Considerando os embriões das espécies nativas, 52,4% foram detectados positivos (11/21). O DNA de Mycoplasma spp. foi detectado em um filhote de Pionus maximiliani morto na eclosão. Testes adicionais dos embriões e das colônias por PCR, com protocolo específico para M. gallisepticum, não revelaram nenhum resultado positivo. As implicações da presença de Mycoplasma na viabilidade embrionária e de filhotes de espécies de aves comerciais ou de conservação são discutidas.

ABSTRACT Industrial broilers raised on helminthic medication-free feed were diagnosed with a severe disease caused by Ascaridia galli, characterized by intestinal hemorrhage and obstruction. A. galli was identified based on the morphological features of the nematode. Broilers were raised for a longer period (63 days) for weight recovery, grouped as stunted (n=500), had low body score and had fetid diarrhea. The duodenum-jejunum segment was the most severely affected with obstruction and had localized accumulation of gas. The intestinal mucosa was severely congested with petechial and suffusive hemorrhages. The outbreak resulted in morbidity of about 10% and mortality of up to 4% and was associated to the absence of preventive medication on feed and slack biosecurity. The reemergence of A. galli is discussed in view of the alternative poultry management and raising conditions for drug free and welfare.

ABSTRACT Serum samples (n=687) from Gallus gallus domesticus were collected for the investigation of antibodies to avian influenza virus (AIV-A) in the family poultry of the surrounding counties of Santa Maria/RS and the metropolitan region of Belo Horizonte/MG, totaling twenty different counties. Additional samples of seventeen (n=17) free-flying ducks (C. moschata pure or hybrid with Anas platyrhynchos) were collected in Belo Horizonte. The chosen tests for the survey were performed as described by the World Organization for Animal Health (OIE), including agar gel immunodiffusion (AGID) for antibodies to AIV-A nucleoprotein (N) and haemagglutination- inhibition (HI) for antibodies to subtype H1. Out of the 704 serum tests performed by AGID, eight (8/704) were revealed positive for antibodies to AIV-A N protein, with six (6/704) retested positive for subtype H1. Two sera tested positive by AGID were shown to be non reactive to the H1 subtype, suggesting specificity to another subtype. A low occurrence of antibodies to influenza A (1.13%) was found, and mostly (75%) specific to subtype H1. This represents an approximately 0,85% overall occurrence for subtype H1 antibodies, with an unknown subtype specific antibodies detected in one free-flying anatid. The low occurrence of antibodies in the family poultry may suggest a low AIV-A activity during the period of study, information which remains to be confirmed by virus detection.

ABSTRACT This study aimed at determining the clinical and pathological effects of the coinfection of young SPF chickens with chicken anemia virus (CAV) and Mycoplasma gallisepticum (MG) vaccine strains. The clinical signs, gross and microscopic lesions were determined after the experimental coinfection broilers with a CAV genotype 1 vaccine strain given intraperitoneally on the first day of age and a MG F-strain vaccine given intranasally on the 8th day of age. The experimental groups included the negative control (group 1), a group infected with the MG F-strain vaccine (group 2), and a group coinfected with CAV and MG vaccines (group 3). Chicks were examined clinically and post mortem at 23 days of age, and gross and microscopic lesions of the trachea, thymus, and air sacs were compared among treatments (Kruskal-Wallis test). Infections were confirmed by PCR for specific genetic fragments of each agent in the target tissues. Mortality was only observed in chicks on group 3, with two deaths and more severe lesions in the trachea, thymus and air sacs compared with groups 1 and 2 (p< 0.01). Dead chicks presented reduced thymus and spleen size, hemorrhagic trachea with catarrhal exudate and partial obstruction, pericarditis, catarrhal airsacculitis, lungs with liquid and ascites. The surviving chicks in group 3 showed more severe respiratory changes than those in group 2, in addition to thymus and spleen size reduction. Results indicate the adverse effects of the coinfection of young chickens with MG F-strain and CAV genotype 1 vaccines.

RESUMO Objetivou-se, no presente experimento, caracterizar os três primeiros jatos da fração rica do ejaculado de jumentos da raça Pêga. Para tal, cinco jumentos foram submetidos à coleta fracionada do sêmen, utilizando-se vagina artificial modelo aberta. Os três primeiros jatos da fração rica do ejaculado foram coletados separadamente e avaliados quanto ao volume, à motilidade e ao vigor espermáticos, à concentração espermática/mL de sêmen e à morfologia espermática. Comparações foram realizadas entre jatos e entre jumentos. Observou-se que o volume do jato diferiu (P<0,05) entre os jumentos, embora fosse similar (P>0,05) entre os jatos. A motilidade espermática não diferiu (P>0,05) entre jatos nem entre jumentos. O vigor espermático foi similar (P>0,05) entre os jatos de um mesmo jumento, e apenas o vigor do jato 1 diferiu (P<0,05) entre os jumentos. Independentemente do jumento, a fração rica do ejaculado foi composta por três jatos apresentando concentrações espermáticas decrescentes (P<0,05), com 955,56; 725,56 e 280,56 x 106 espermatozoides/mL de sêmen. A morfologia espermática diferiu entre os três jatos avaliados apenas para a incidência de defeitos de peça intermediária, sendo maior no jato três (4,26%), em relação aos jatos um (3,36%) e dois (3,38%). Comparando-se as características morfológicas do sêmen entre os cinco jumentos avaliados, independentemente do jato, observaram-se diferenças entre os reprodutores quanto ao percentual de espermatozoides normais, com gota citoplasmática proximal, com defeitos de peça intermediária e de cabeça.

ABSTRACT The first three jets of the sperm-rich fraction of Pêga jackasses were collected and assessed separately. Five fertile Pêga jackasses were used as semen donors and underwent fractionated semen collection, using an open model artificial vagina. The first three jets of the semen were collected separately and assessed for volume, sperm motility, vigor, concentration/mL of semen, and sperm morphology. These characteristics were compared between first, second and third jets and between jackasses. It was observed that the jet volume differed (P<0.05) between jackasses, although it was similar (P>0.05) between first, second and third jets. Sperm motility did not differ (P>0.05) between jets and jackasses. Vigor was similar (P>0.05) between jets of the same jackass, and only the first jet differed (P<0.05) between jackasses. The first, second and third jets of the sperm-rich fraction had decreased sperm concentrations (P<0.05) of 955.56, 725.56 and 280.56x 106 sperm/mL of semen, respectively. Sperm morphology differed between the first three jets only for the incidence of mid-piece defect, higher in the third one (4.26%), compared to the first (3.36%) and second (3.38%) ones. When comparing the morphological characteristics of the sperm-rich fraction between five jackasses, regardless of the jet, there were differences in the percentage of normal sperm, proximal cytoplasmic droplet, mid-piece and head defects.

ABSTRACT The incidence of the psittacine beak and feather disease virus (BFDV) was investigated in Brazilian native parrots with normal feathering arriving at rescue and triage centers for wild animals (CETAS, IBAMA) in the state of Minas Gerais, Brazil. BFDV DNA was investigated by previously described PCR technique for the partial amplification of BFDV ORF-1 in DNA extracts from blood, cloacal swab or liver of psittacines. Some birds provided more than one sample. Nine species of psittacines were sampled between January 2009 and October 2010. Blood (n=46) or cloacal swab (n=128) samples were obtained from psittacines immediately upon arrival at the triage centers. Liver samples were collected from necropsied birds dead on arrival (n=167). All swab samples were negative, except for one Ara ararauna individual (n=3) which blood presented the BFDV DNA. On the other hand, 11 liver samples were positive for BFDV DNA, with a prevalence of 7.8% in Amazona aestiva (n=140). No BFDV DNA was detected in the liver of Amazona amazonica (n=11), A. vinacea (n=5), A. rhodochorytha (n=4), Anodorhynchus hyacinthinus (n=3), Ara ararauna, (n=3), Aratinga leucophtalma (n=2), Guarouba guarouba (n=1) and Pionus maximiliani (n=1). In most cases, alopecia was not associated with BFDV detection in liver, and liver histopathology was inconclusive. Although all cloacal swab samples were negative, a few psittacines (n=19) that died at CETAS-Belo Horizonte were retested, and 21% were detected as positive in liver. A group of psittacines (n=16) was clinically evaluated, and despite showing feather dystrophy, all birds were negative in the cloacal swabs, except for one, which blood sample was positive (A. ararauna). The obtained sequences of the BFDV strains BH 215 and BH 732 were deposited in the GenBank (JQ649409 and JQ649410). A 98% similarity with strain sequences described in Australia, Japan, and New Zealand was observed. It is possible that these strains arrived in Brazil through the legal and illegal trade of parrots. However, it was not possible to associate BFDV infection with the geographical origin of birds and no local marker was detected. The rates of detection, although similar to other studies, indicate the tendency of a high incidence of the disease, possibly associated with stress, and high bird density and wide transmission in captivity conditions.

AbstractBiosurfactants have a wide range of applications in emulsions, separations, and solubilization because these chemicals reduce the surface tension and viscosity of solutions. This work studied rhamnolipid production using a batch bioreactor with a working volume of 1.5 liters, Pseudomonas aeruginosa ATCC (American Type Culture Collection) 10145 strain, and the greasy effluent from the slaughter of poultry and pigs as the substrate. The main goal of this research was to evaluate the level of aeration, agitation speed and inoculum concentration using a Central Composite Design (CCD). Experimental conditions were selected using the surface response technique obtained from the CCD, and the results were validated to test the reproducibility. The following operating conditions were selected: 1.2 vvm level of aeration, 600 rpm agitation speed, and 1.0 g/L biomass inoculum concentration. Under these conditions, the following results were obtained: the rhamnose production, surface tension and emulsifying index were 5.37 g/L, 25.6 dyne/cm and 100%, respectively.

Descreve-se a ocorrência de Aviadenovirus (FAdV) na avicultura mineira. Foram amostrados fígados de poedeiras jovens (n=25) e velhas (n=25) e de frangos de corte (n=300). Em matrizes pesadas foram amostrados fígados (n=25) e fezes (n=25). O estudo envolveu as principais regiões avícolas do Estado de Minas Gerais. O DNA de FAdV foi pesquisado por PCR universal, descrito para a amplificação do gene que codifica o hexon de Aviadenovirus, usando FAdV Phelps como referência. Foi demonstrada a presença do DNA de FAdV em 100% (25/25) das poedeiras velhas (78 semanas de idade) e em 36% (9/25) das jovens (18 dias). Em frangos de corte, a detecção variou entre 24 e 86%. Embora as fezes das matrizes tenham sido negativas, foi obtido o amplicon específico em 4/25 dos fígados dessas mesmas aves, indicando menor sensibilidade para detecção nas fezes. Em amostras da avicultura familiar (fígado), colhidas de uma das regiões de avicultura intensificada, foi detectado o genoma de FAdV em 100% das galinhas (n=12). A constatação de alta disseminação de FAdV em aves da avicultura industrial e familiar de Minas Gerais contribui para o entendimento da epidemiologia de Aviadenovirus. As sequências nucleotídicas dos produtos de PCR alinharam com FAdV da espécie D de Aviadenovirus. A demonstração de FAdV em reprodutores indica risco de transmissão vertical e reforça a necessidade de biosseguridade estrita nesses plantéis. A presença de FAdV em diversos setores avícolas, incluindo poedeiras comerciais, frangos de corte, reprodutores e galinhas da avicultura familiar, recomenda a biosseguridade estrita entre as criações de mesmo tipo e de tipos diferentes de aves. A detecção em matrizes pode indicar a continuada geração de progênies infectadas.

The occurrence of Aviadenovirus (FAdV) was investigated in chickens from the poultry industry of Minas Gerais state, Brazil. The investigation was conducted due to the scarcity of recent data in the country and its description in neighboring countries. For this purpose, livers were collected from layer chicks (n=25), older layers (n=25), broilers (n=300), and livers (n=25) and stool (n=25) samples from broiler breeders, representing the major poultry regions of the state. FAdV DNA was demonstrated using a previously described PCR protocol for amplifying part of the hexon gene encoding sequence. FAdV was found in layer chicks (36%), widespread (100%) in older layers, and with regional differences in broilers (24-86%). Although all broiler breeder stools were negative, FAdV DNA was detected in livers (16%, 4/25) of stool-negative birds. In order to obtain additional information on the circulation of the infection, livers of subsistence chickens collected from one poultry intensive region, were evaluated (n = 12), with FAdV being detected in all samples. FAdV was found in young and old layers, broilers, broiler breeders and free-range chickens, and results suggest the circulation of FAdV among different types of chickens. The detection in older layer chickens may indicate an extended risk of horizontal transmission in regions of Minas Gerais with mixed activity of egg and meat type chickens and poor biosecurity strategies. The infection in breeders may indicate vertical transmission and the continuous production of infected progenies. The hexon-gene-targeted PCR amplicon sequences aligned with FAdV of species D of Aviadenovirus. Results indicate the necessity for biosecurity, especially for breeders, separating flocks according to origin, age and health status, which will be an advantage regarding any pathogen.

Avaliou-se a ocorrência de Orthoreovirus (ARV) e Rotavirus (AvRV) aviários na avicultura industrial de Minas Gerais. Foram colhidas cinquenta e quatro amostras de fezes de frangos de corte entre um e 45 dias e de frangas de postura de 10 a 13 semanas de idade. Para análise de ARV, o RNA foi imediatamente extraído (Trizol), transcrito em cDNA e avaliado em uma PCR com oligonucleotídeos iniciadores específicos para ARV. Para a investigação de AvRV, os extratos de RNA foram obtidos por fenol-clorofórmio e submetidos à eletroforese em gel de poliacrilamida. Todas as amostras foram também avaliadas para o DNA do vírus da anemia das galinhas (CAV) em uma nested-PCR específica. Em frangos de corte, a positividade encontrada para ARV foi de 5,55% e para AvRV de 9,25%. CAV foi detectado em coinfecção em um plantel com refugagem, claudicação e prostração. Nenhuma amostra de poedeiras foi positiva para ARV ou AvRV. Material de plantel com sinais clínicos foi purificado e inoculado em ovos SPF embrionados, sendo obtidas lesões hemorrágicas e focos brancos na membrana cório-alantóide. O sequenciamento dos produtos de PCR e de embrião agrupou os isolados de ARV com a estirpe S1133, historicamente usada como vacina viva. Os resultados sugerem a continuada circulação da infecção por estirpes assemelhadas a ARV S1133 nas regiões de avicultura industrial. Os índices de detecção de ARV, AvRV e CAV indicam que a intensificação nas regiões produtoras tem resultado em falhas de biosseguridade.

Fifty-four fecal samples taken from broiler chickens from 1 to 45 days of age, and of pullets from 10 to 13 weeks of age, original from eight different poultry regions in the state of Minas Gerais, Brazil, were collected from March 2008 to January 2010 for avian Orthoreovirus (ARV) and avian Rotavirus (AvRV) analyses. For the assay of ARV, RNA was immediately extracted (Trizolâ) and transcribed into cDNA for assaying in a nested-PCR with ARV-specific primers. For AvRV, polyacrylamide gel electrophoresis (PAGE) was performed with RNA extracts obtained by phenol-chloroform extraction. CAV was additionally investigated through a nested-PCR of thymus and spleen. Results found 5.55% positive for ARV and 9.25% for AvRV. Also, CAV and ARV genomes were detected in co-infection, in a highly prostrated and claudicating chicken flock. No ARV or AvRV infections were detected in pullets. Material of a clinically affected flock was inoculated into SPF embryos, resulting in embryonic hemorrhage, whitish foci in the chorio-allantoic membrane and death. Sequencing of ARV amplicons and isolate cDNA grouped local strains with the ARV S1133 strain, historically used in live vaccines, suggesting the continued circulation of this vaccine virus strain in intensive poultry regions. Detection rates for ARV and AvRV, as well as the presence of CAV, were additionally indicative of failing biosecurity strategies for the intensive poultry regions examined.

Vacinas avícolas vivas comerciais produzidas entre 1991 e 2005 foram examinadas para a presença de genomas dos vírus da anemia infecciosa das galinhas (Gyrovirus CAV), da hepatite por corpúsculo de inclusão (Aviadenovirus FAdV) e da artrite viral/síndrome da má absorção (Orthoreovirus aviário ARV). Vinte e seis partidas de vacinas vivas liofilizadas de oito fabricantes com lacre original foram examinadas. As extrações de DNA e PCR de CAV e FAdV, e de RNA e RT-PCR para ARV, foram descritas previamente. Contaminações triplas de ARV, CAV e FAdV foram detectadas em vacinas de mesmo fabricante, produzidas em 1991 e 1992 contra a doença de Newcastle (DN), e para a encefalomielite aviária, produzida em 1994. ARV e CAV em co-infecção foram encontrados em vacinas contra a doença de Marek liofilizadas produzidas em 1996 por dois fabricantes diferentes. Genoma de ARV foi detectado em vacinas contra a bronquite infecciosa de setembro e dezembro de 1998, doença infecciosa bursal, de dezembro de 1998 e DN de janeiro de 1998. Três dos oito fabricantes apresentaram vacinas com contaminação e cinco nunca apresentaram vacinas contaminadas. Nenhuma vacina produzida a partir de 2001 apresentou contaminação. Cogita-se um papel epidemiológico para vacinas vivas, como fonte de infecção para ARV, CAV e FAdV e, potencialmente determinante da atual alta disseminação destes.

Nested PCR foi utilizada para o diagnóstico de encefalite bovina por herpesvírus incluindo o herpesvírus bovino 5 (BHV-5), o herpesvírus bovino 1 (BHV-1), o vírus da doença de Aujeszky (SHV-1) e o herpesvírus ovino 2 (OHV-2) em 14 fragmentos do sistema nervoso central (SNC) de bovinos que morreram com sinais neurológicos. Embora o BHV-4 não seja reconhecido como vírus neurotrófico, foi detectado nos casos de encefalite que ocorreram isoladamente em diferentes épocas do ano e não apresentaram nenhuma sazonalidade. A duração do curso clínico variou entre 1 e 15 dias, e em 64,3% dos casos manifestou-se entre 1 e 2 dias. Os sinais neurológicos mais freqüentemente observados foram ataxia, apatia, instabilidade, opistótono, movimentos de pedalagem, nistagmo e sialorréia. Nos ensaios de PCR nested realizados a partir do DNA obtido do SNC, não foi encontrado evidência de: BHV-1, SHV-1 ou OHV-2 em nenhuma das amostras. Mas, a presença de BHV-4 foi encontrada em todos os fragmentos do SNC de bovinos que morreram com sinais neurológicos. Além disso, o BHV-5 foi encontrado em associação com o BHV-4 em duas dessas amostras.

A nested PCR assay was used to diagnose bovine encephalitis through herpesviruses including bovine herpesvirus 5 (BHV-5), bovine herpesvirus 1 (BHV-1), Aujeszky's disease virus (SHV-1), and ovine herpesvirus 2 (OHV-2) in 14 fragments of central nervous system (CNS) from cattle that died with neurological signs. In addition, as some samples of bovine herpesvirus type 4 (BHV-4) have been isolated from neural tissue, it was also tested by nested PCR. The cases of encephalitis occurred in isolation at different times of the year and did not present any seasonality. The duration of the clinical course ranged between 1 to 15 days, and in 64.3% of the cases it manifested between 1 to 2 days. The most frequently observed neurological signs were ataxia, recumbency, unsteadiness and inability to stand, opisthotonus, paddling movements, nystagmus and ptyalism. In the nested assay, there was no evidence of: BHV-1, SHV-1 or OHV-2 in the DNA obtained from the CNS in any of the samples. But the presence of BHV-4 was found in all fragments of the CNS in cattle which died presenting neurological signs. Moreover, BHV-5 was found in association with BHV-4 in two of these samples.

Blood samples from 1,072 domestic cats of nine administrative regions of Belo Horizonte, MG, were collected and tested using PCR nested for the occurrence of feline leukemia virus (FeLV). Overall occurrence was 47.5% (507/1072) being North (68.1%) and East (54.4%) the most prevalent areas. Epidemiological data showed that FeLV infection was very common among examined cats and breed neither gender nor were predisposing factors for FeLV. The results suggest that the agglomeration of a large number of cats in the same environment can be an important factor for the increase in the rate of transmission of this retrovirus among domestic cats in the studied city.

Em agosto de 2008, um garanhão da raça Mangalarga Marchador, de quatro anos de idade, com histórico clínico de apatia, inapetência e edema de prepúcio e escroto, apresentou, ao exame clínico, exsudato purulento fluindo pelo óstio prepucial, prepúcio e mucosa peniana com inúmeras lesões circulares de bordos elevados e hiperêmicos, centro ulcerado recoberto por material amarelado de aspecto fibrinoso, com distribuição multifocal. Histologicamente, a mucosa peniana apresentou áreas de ulceração associadas a infiltrado inflamatório misto, com necrose multifocal e moderado acúmulo de fibrina, que se estendiam para o tecido conjuntivo adjacente. O diagnóstico morfológico foi de balanopostite ulcerativa fibrino-necrótica multifocal intensa, similar ao encontrado em casos de exantema coital equino (ECE), causado pelo herpesvírus equino 3 (EHV-3). Amostra de pele do prepúcio e sangue, colhido em EDTA, foram submetidos a ensaios de PCR específicos para EHV-3, observando-se a amplificação de um produto de tamanho esperado de 518pb. A detecção do EHV-3 foi confirmada por meio de seu sequenciamento, sendo a sequência de nucleotídeos depositada no GenBank sob o número GQ336877. As sequências de nucleotídeos e as de aminoácidos deduzidos apresentaram identidade de 99% e 100%, respectivamente, com a sequência de EHV-3 disponível no GenBank, número AF081188. Após 15 dias de tratamento, houve completa cicatrização das lesões, com despigmentação da pele, principalmente, no prepúcio e na bolsa escrotal. Com base nos achados clínicos, histopatológicos, PCR e sequenciamento, concluiu-se tratar de um caso de exantema coital equino, sendo o primeiro com confirmação definitiva do agente etiológico no Brasil.

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.

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.