Molecular detection of Mycoplasma gallisepticum in different poultry breeds of Abbottabad and Rawalpindi, Pakistan / Detecção molecular de Mycoplasma gallisepticum em diferentes raças de aves de Abbottabad e Rawalpindi, Paquistão

Abstract The poultry sector in Pakistan is contributing mainly in bridging gap between demand and supply for protein. Mycoplasma gallisepticum is an emerging bacterium causing serious problems in poultry industry of Pakistan. A cross-sectional study was conducted to evaluate the M. gallisepticum load in poultry populated regions of Pakistan. Total 600 serum and 600 swab samples were collected, 200 from each broiler, layers and breeders poultry in Rawalpindi and Abbottabad districts. Serum samples were analyzed through ELISA for seroprevalence. Swabs were cultured on Frey's medium followed by PCR and partial mgc2 gene sequencing. Results of seroprevalence of M. gallisepticum showed that layers (75%, n=150) are more positive as compared to breeders (70%, n=140) and broilers (50%, n=100). Typical colonies of the M. gallisepticum were observed in breeder (26.5%), followed by layer (21%) and broilers (9%). A total of 37.1% (n=42) samples were identified positive through PCR out of total 113 cultured based positive samples. A total of six M. gallisepticum isolates of current study showed 98-99 percent similarity with previously reported isolates on the basis of mgc2 gene partial sequencing. The M. gallisepticum was found highly prevalent in different poultry breads. Results of this study would add into basic data and provide a direction for livestock sector to strengthen a control strategy for mycoplasmosis in poultry farms.

Resumo O setor avícola do Paquistão está contribuindo principalmente para preencher a lacuna entre a demanda e a oferta de proteína. Mycoplasma gallisepticum é uma bactéria emergente que causa sérios problemas na indústria avícola do Paquistão. Um estudo transversal foi conduzido para avaliar a carga de M. gallisepticum em regiões de avicultura do Paquistão. Um total de 600 amostras de soro e 600 amostras de esfregaço foi coletado, 200 de cada frango de corte, poedeiras e aves reprodutoras nos distritos de Rawalpindi e Abbottabad. Amostras de soro foram analisadas por ELISA para soroprevalência. As zaragatoas foram cultivadas em meio Frey, seguido de PCR e sequenciação parcial do gene mgc2. Os resultados da soroprevalência de M. gallisepticum mostraram que as poedeiras (75%, n = 150) são mais positivas em comparação com matrizes (70%, n = 140) e frangos de corte (50%, n = 100). Colônias típicas de M. gallisepticum foram observadas em reprodutoras (26,5%), seguidas de poedeiras (21%) e frangos de corte (9%). Um total de 37,1% (n = 42) das amostras foi identificado como positivas por PCR de um total de 113 amostras positivas baseadas em cultura. Um total de seis isolados de M. gallisepticum do estudo atual mostrou 98-99% de similaridade com isolados relatados anteriormente com base no sequenciamento parcial do gene mgc2. O M. gallisepticum foi encontrado com alta prevalência em diferentes pães de aves. Os resultados deste estudo acrescentariam dados básicos e forneceriam orientação para o setor pecuário fortalecer uma estratégia de controle da micoplasmose em granjas avícolas.

A common Salmonella Enteritidis sequence type from poultry and human gastroenteritis in Ibagué, Colombia / Un tipo de secuencia común de Salmonella Enteritidis de origen aviar y de humano con gastroenteritis en Ibagué, Colombia

Abstract Introduction: Salmonella Enteritidis is a major cause of human salmonellosis in the world, with contaminated eggs and raw chicken meat as the main routes of infection. The main Salmonella spp. serovars circulating in laying hen farms, the surface of eggs, and in raw chicken carcasses have been identified in Ibagué, Colombia. However, it is unknown whether those serovars are responsible for human gastroenteritis. Objective: To evaluate the genetic relationship between gastroenteritis and Salmonella Enteritidis isolates from poultry and humans using multilocus sequence typing (MLST). Materials and methods: Salmonella spp. was isolated from clinical cases of gastroenteritis (n=110). Antibiotic susceptibility tests, followed by serotyping and MLST were conducted and S. Enteritidis was compared to those from laying hen farms and marketed eggs. Results: Ten isolates of Salmonella spp. were obtained from the stools of people with gastroenteritis. The prevalence of Salmonella spp. in human stools was 9.09%, and S. Enteritidis (n=4), S. Typhymurium (n=2), S. Newport (n=1), S. Uganda (n=1), S. Grupensis (n=1), and S. Braenderup (n=1) were the main serotypes. MLST indicated that a common S. Enteritidis sequence type (ST11) was present in all three sources and showed the same antibiotic resistance pattern. Conclusion: Salmonella Enteritidis ST11 constitutes a link between consumption and manipulation of contaminated eggs and human gastroenteritis in Ibagué. Additional studies would be required to establish if other Salmonella serovars isolated from raw chicken meat are also associated with human gastroenteritis.

Resumen Introducción. Salmonella Enteritidis es una de las mayores causas de salmonelosis en el mundo; los huevos contaminados y la carne de pollo cruda son sus principales fuentes de infección. En Ibagué, Colombia, se han identificado los principales serovares que circulan en granjas, superficies de huevos y canales de pollo, pero se desconoce si esos serovares son responsables de la gastroenteritis. Objetivo. Evaluar la relación genética entre los aislamientos de Salmonella Enteritidis de aves de corral y de humanos con la gastroenteritis mediante tipificación de multiloci de secuencias (Multilocus Sequence Typing, MLST). Materiales y métodos. Se aisló Salmonella spp. de casos clínicos de gastroenteritis (n=110). Se hizo la prueba de sensibilidad antibiótica, así como la serotipificación y la tipificación mediante MLST, y se comparó S. Enteritidis de humanos con la hallada en granjas de gallinas ponedoras y en huevo comercializado (n=6). Resultados. Se aislaron 10 cepas de Salmonella spp. a partir de heces de humanos con gastroenteritis. Se obtuvo una prevalencia de Salmonella spp. de 9,09%, y se identificaron los serotipos S. Enteritidis (n=4), S. Typhymurium (n=2), S. Newport (n=1), S. Grupensis (n=1), S. Uganda (n=1) y S. Braenderup presentes en pacientes con gastroenteritis. Mediante la MLST, se comprobó que un tipo de secuencia común (ST11) de S. Enteritidis estuvo presente en todas las tres fuentes y presentó el mismo patrón de resistencia antibiótica. Conclusión. Salmonella Enteritidis ST11 constituye un vínculo entre el consumo y la manipulación de huevos contaminados, y la gastroenteritis en humanos en Ibagué. Se requieren estudios complementarios para conocer si otros serovares de Salmonella aislados de carne de pollo cruda también se asocian con la gastroenteritis en humanos.

The first study of molecular prevalence and species characterization of Cryptosporidium in free-range chicken (Gallus gallus domesticus) from Brazil / Primeiro estudo de prevalência molecular e caracterização de espécies de Cryptosporidium em galinhas (Gallus gallus domesticus) colonial/caipiras do Brasil

Abstract Rearing free-range chicken is based on grazing feeding patterns, and these animals could be potential environmental contaminants of Cryptosporidium oocysts for humans and other animals. Therefore, the present study aimed to evaluate the molecular prevalence of Cryptosporidium spp. in free-range chickens from Brazil. A total of 351 fecal samples from chickens were examined from 20 farms. For detection of Cryptosporidium spp., 18S rRNA gene fragments were amplified using a nested PCR reaction. Positive samples were sent for sequencing. The overall prevalence of Cryptosporidium was 25.6% (95% CI = 21.2% - 30.6%). Sequencing of the amplified fragments allowed for the identification of three species: C. meleagridis in 57 (62.6%), C. baileyi in 15 (16.4%), C. parvum in 3 (3.2%) samples, and a new Cryptosporidium genotype (C. genotype BrPR1) in 3 (3.2%) samples. Cryptosporidium genotype BrPR1 has not yet been classified as a species, and its host spectrum is not known. Cryptosporidium, including zoonotic species, exists at a high prevalence in free-range chickens within the region studied.

Resumo A criação de galinhas no estilo colonial/caipira é baseada em padrões de alimentação de pastagem, o que as torna potenciais contaminantes ambientais de oocistos de Cryptosporidium para humanos e outros animais. Portanto, o presente estudo teve como objetivo avaliar a prevalência molecular de Cryptosporidium spp. em galinhas criadas em sistema colonial/caipira. Um total de 351 amostras de fezes de frangos foram examinadas em 20 fazendas. Para a detecção de Cryptosporidium spp., os fragmentos do gene rRNA 18S foram amplificados utilizando-se a reação de nested-PCR. A prevalência global de Cryposporidium foi de 25,6% (IC 95% = 21,2% - 30,6%). O sequenciamento dos fragmentos amplificados permitiu a identificação de três espécies que infectam aves: C. meleagridis em 57 (62,6%), C. baileyi em 15 (16,4%), C. parvum em 3 (3,2%) amostras, bem como, um novo genótipo de Cryptosporidium (C. genótipo BrPR1) foi identificado em 3 (3,2%) amostras. Cryptosporidium genotipo BrPR1 não foi ainda classificado como uma espécie, e seu espectro de hospedeiros é desconhecido. O presente trabalho permitiu concluir que Cryptosporidium, incluindo espécies zoonóticas, existe com alta prevalência em galinhas criadas em sistema colonial/caipira na região estudada.

Epidemiology of Megninia spp. in laying flocks from the State of Minas Gerais, Brazil / Epidemiologia de Megninia spp. em estabelecimentos de postura no Estado de Minas Gerais, Brasil

Aspects related to the epidemiology of Megninia ginglymura were studied, in order to determine predisposing factors for the ocurrence in layer chicken houses of commercial farms in the State of Minas Gerais, Brazil. An observational, cross-sectional, analytical survey was conducted for identifying major risks or protection factors for infestations. Four hundred thirty-one houses located in forty-three farms were visited in 2012. Regarding the farms, 17 (39.53%) had chickens infested by M. ginglymura and 1 (2.32%) presented Megninia cubitalis. The epidemiological aspects of the occurrence of M. ginglymura were investigated using multiple logistic regression models. The chicken house organization was an important protection factor involved in the occurrence of M. ginglymura (odds ratio [OR] = 0.85). The presence of subsistence breedings of poultry within the premises or at the proximity of a farm was characterized as a risk factor for the occurrence of infestations (OR = 3.92). The use of mineral oil (OR = 0.16) was considered protective against new infestations. Some regions of the state were characterized by a higher risk for mite infestation than others.

Aspectos relacionados à epidemiologia de infestações pelo ácaro Megninia ginglymura foram estudados com o objetivo de determinar os principais fatores para a ocorrência desse ectoparasito em galinhas poedeiras de granjas comerciais no Estado de Minas Gerais, Brasil. Um estudo observacional, analítico e seccional foi realizado para verificar os principais fatores de risco ou proteção para as infestações. Quatrocentos e trinta e um galpões distribuídos em quarenta e três granjas foram visitados, no ano de 2012. Em relação às granjas, 17 (39,53%) possuíam aves infestadas por M. ginglymura e uma (2,32%) apresentou galinhas infestadas por Megninia cubitalis. Os aspectos epidemiológicos para a ocorrência de M. ginglymura foram investigados, utilizando-se modelos de regressão logística múltipla. A organização dos galpões nas granjas foi verificada como um importante fator de proteção envolvido na epidemiologia dessa espécie (Razão de Chances=0,85). A presença de criações de galinhas de subsistência dentro das instalações das granjas ou nas proximidades (Razão de Chances =3,92) foi caracterizada como um fator de risco para a ocorrência de infestações. O uso de óleo mineral foi considerado um fator de proteção contra infestações por M. ginglymura. Algumas regiões do Estado de Minas Gerais foram caracterizadas como de maior risco do que outras para infestações por esse ácaro.

Dissimilarity of ccrAB gene sequences between methicillin-resistant Staphylococcus epidermidis and methicillin-resistant Staphylococcus aureus among bovine isolates in Korea

The sequences of the ccrAB genes from bovine-, canine- and chicken-originating methicillin-resistant Staphylococcus (S.) epidermidis (MRSE) and bovine methicillin-resistant Staphylococcus (S.) aureus (MRSA) were compared to investigate the frequency of intra-species horizontal transfer of the staphylococcal cassette chromosome mec (SCCmec) complex. Nineteen MRSE strains were isolated from bovine milk, chickens, and dogs, and their genetic characteristics were investigated by multilocus sequence typing and SCCmec typing. Among the animal MRSE strains, the most frequent SCCmec type was type IV, which consisted of the type B mec complex and ccrAB type 2. The ccrA2 and ccrB2 genes were sequenced from the bovine, chicken and canine MRSE strains and compared with those of the bovine MRSA strains. The sequences generally clustered as MRSA and MRSE groups, regardless of the animal source. Additionally, no bovine MRSE sequence was associated with the bovine MRSA groups. Although most of the bovine MRSE and MRSA isolates possessed SCCmec type IV sequences, our results suggest that the intra-species gene transfer of the SCCmec complex between bovine S. aureus and bovine S. epidermidis strains is not a frequent event.

Pathotyping avian pathogenic Escherichia coli strains in Korea

To examine the genetic background of avian pathogenic Escherichia coli (APEC) that affects virulence of this microorganism, we characterized the virulence genes of 101 APEC strains isolated from infected chickens between 1985~2005. Serotypes were determined with available anti-sera and median lethal doses were determined in subcutaneously inoculated chicks. The virulence genes we tested included ones encoding type 1 fimbriae (fimC), iron uptake-related (iroN, irp2, iucD, and fyuA), toxins (lt, st, stx1, stx2, and vat), and other factors (tsh, hlyF, ompT, and iss). Twenty-eight strains were found to be O1 (2.0%), O18 (3.0%), O20 (1.0%), O78 (19.8%), and O115 (2.0%) serotypes. The iroN (100%) gene was observed most frequently followed by ompT (94.1%), fimC (90.1%), hlyF (87.1%), iss (78.2%), iucD (73.3%), tsh (61.4%), fyuA (44.6%), and irp2 (43.6%). The strains were negative for all toxin genes except for vat (10.9%). All the strains were classified into 27 molecular pathotypes (MPs). The MP25, MP19, and MP10 pathotypes possessing iroN-fimC-ompT-hlyF-iucD-tsh-iss-irp2-fyuA (22.8%), iroN-fimC-ompT-hlyF-iucD-tsh-iss (21.8%), and iroN-fimC-ompT-hlyF-iss (11.9%) genotypes, respectively, were predominant. Redundancy of iron uptake-related genes was clearly observed and some strains were associated with higher mortality than others. Therefore, strains with the predominant genotypes can be used for diagnosis and vaccine.

Coccidiosis in broiler chickens raised in the Araguaína region, State of Tocantins, Brazil / Coccidiose em frangos de corte criados na região de Araguaína, estado do Tocantis, Brasil

Eimeriosis is responsible for causing serious problems in poultry, mainly characterized by reduced weight gain and abnormalities of food conversion efficiency, thereby causing great economic losses. The aim of this study was to evaluate the epidemiology of eimeriosis in broiler chickens in the Araguaina region, State of Tocantins, Brazil. Samples from five farm properties were collected and sent to the Hygiene and Public Health Laboratory, School of Veterinary Medicine and Zootechnics, Federal University of Tocantins. From the parasitological analysis, it was shown that all the properties examined were positive for Eimeria species. 63.1 percent of the sheds were positive, with findings of oocysts of E. maxima, E. acervulina, E. mitis and E. tenella. It was concluded that all properties evaluated were positive for four species of the genus Eimeria, thus demonstrating that the sanitary strategies followed in poultry rearing had flaws that allowed pathogens to spread in poultry pens.

A eimeriose é responsável por causar graves problemas avícolas caracterizados, principalmente, por redução do ganho de peso e alteração na conversão alimentar, gerando grandes perdas econômicas. O objetivo deste trabalho foi avaliar a epidemiologia da eimeriose em frangos de corte na região de Araguaína, Estado do Tocantins, Brasil. Amostras de cinco propriedades foram colhidas e encaminhadas ao Laboratório de Higiene e Saúde Pública da Escola de Medicina Veterinária e Zootecnia da Universidade Federal do Tocantins. Após análise parasitológica, comprovou-se que todas as propriedades avaliadas apresentaram positividade para espécies do gênero Eimeria, tendo 63,1 por cento dos galpões positivos, sendo encontrados oocistos de E. maxima, E. acervulina, E. mitis e E. tenella. Conclui-se que todas as propriedades avaliadas, apresentaram positividade para quatro espécies do gênero Eimeria comprovando que as estratégias sanitárias adotadas nas criações apresentam falhas que possibilitam a disseminação do patógeno nos lotes avícolas.

Detection & characterization of Shiga toxin producing Escherichia coli (STEC) & enteropathogenic Escherichia coli (EPEC) in poultry birds with diarrhoea.

Background & objectives Limited information is available on shiga toxin producing Escherichia coli (STEC) in animals and birds from India. An outbreak of acute diarrhoea in poultry birds at Aizawl, Mizoram was investigated for detection and characterization of STEC and enteropathogenic E. coli (EPEC). Methods E. coli was isolated and identified from rectal swabs, intestinal contents, heart blood and spleen of 19 poultry birds that died due to acute diarrhoea during the outbreak. Phenotypic characterization was done by standard bacteriological and biochemical techniques. All the isolates were serotyped based on their somatic antigens. Virulence genes (stx1, stx2, eaeA and hlyA) were detected by multiplex PCR assay. Results A total of 42 E. coli isolates were obtained, of which 24 belonged to 3 serogroups (O64, O89 and O91) and the remaining 18 were untypable (UT). Altogether, 14 (33.33%) isolates carried at least 1 virulence gene, of which 10 (23.81%) and 4 (9.52%) were recorded as STEC and EPEC, respectively. Of the 10 STEC isolates, one carried only stx2, one carried stx2 and hlyA, four carried stx1, stx2 and hlyA, two carried stx1, eaeA and hlyA genes and two carried stx1 and eaeA. Of the four EPEC isolates, two carried eaeA and hlyA, one carried only eaeA gene and 1 carried only hlyA gene. Interpretation & conclusions This is the first report on the involvement of STEC in poultry in India.

Anticorpos contra vírus em galinhas de terreiro do Estado do Rio Grande do Sul, Brasil / Antibodies against avian viruses in backyard poultry from the Rio Grande do Sul State, Brazil

No Brasil, a população de aves conhecida como galinhas de terreiro encontra-se fora do sistema de biosseguridade aplicada às criações comerciais. Para investigar a presença de anticorpos contra alguns vírus específicos nesta população, foram coletadas amostras de sangue de 867 aves não-vacinadas em 60 propriedades de 22 municípios do Estado do Rio Grande do Sul, Brasil. O soro foi testado para a presença de anticorpos contra o vírus da bronquite infecciosa das galinhas (IBV), reovírus aviário (ARV) e o vírus da doença infecciosa da bolsa (IBDV) pela técnica de soroneutralização. Anticorpos contra IBV foram detectados em 65 por cento (564/867) das amostras, contra ARV em 21,6 por cento (187/867) e contra IBDV em 80,2 por cento (695/867) das aves. Todas as propriedades avaliadas apresentavam uma ave positiva para anticorpos contra IBV e IBDV e 88,3 por cento delas eram positivas para ARV. Os resultados demonstram que esses vírus estão presentes em galinhas de terreiro nas criações avícolas não-industriais da região central do Estado. Os resultados indicam a necessidade de um programa de vigilância permanente nessa população e ainda indicam a necessidade de avaliar o impacto destas infecções nos próprios plantéis e o risco associado à transmissão destas às criações comerciais.

The backyard poultry are not included in the biosecurity system applied in commercial flocks in Brazil. To investigate the presence of antibodies to specific viral pathogens in this population, blood samples were collected from 867 non-vaccinated birds, from 60 flocks in 22 counties of the Rio Grande do Sul State, Brazil. The samples were tested to detect antibodies against infectious bronchitis virus (IBV), avian reovirus (ARV) and infectious bursal disease virus (IBDV), through the virus neutralization test. Antibodies to IBV were detected in 65 percent (564/867), against ARV in 21.6 percent (187/867), and against IBDV in 80.2 percent (695/867) of the samples. All the flocks had chickens positive to IBV and IBDV antibodies, and 88.3 percent of them harbored antibodies to ARV. The results show the presence of these viruses in backyard poultry from the central region of the State. It also indicates the need for additional studies aimed to evaluate the real importance of these infections for this type of flocks.

Antibiotic-resistant Salmonella spp. from human and non-human sources in Oman

We investigated the antibiotic resistance of various strains of Salmonella isolated from different sources in Oman, namely sewage water, chicken and food handlers. Of 1242 isolates from food handlers, 15 [1.2%] were resistant to one or more antibiotic. Of these, 41% were resistant to ampicillin, 42% to co-trimoxazole and 22% to chloramphenicol. None of the isolates was resistant to ciprofloxacin. Of 515 isolates from chicken and 432 from sewage water, 23.7% and 14.1% respectively were resistant to one or more antibiotic. Susceptibility to the different antibiotics varied but none of the isolates from chicken was resistant to amikacin

Antimicrobial susceptibility and molecular detection of chloramphenicol and florfenicol resistance among Escherichia coli isolates from diseased chickens

Seventy Escherichia coli isolates recovered from diseasedchickens diagnosed with colibacillosis in Henan Province,China, between 2004 and 2005 were characterized forantimicrobial susceptibility profiles via a broth doublingdilution method. Overall, the isolates displayed resistanceto trimethoprim-sulfamethoxazole (100%), oxytetracycline(100%), ampicillin (83%), enrofloxacin (83%), and ciprofloxacin(81%), respectively. Among the phenicols, resistance wasapproximately 79% and 29% for chloramphenicol andflorfenicol, respectively. Molecular detection revealed thatthe incidence rates of the floR, cmlA, cat1, cat2 and cat3were 29, 31, 16, 13, and 0%, respectively. Additionally,10% of the isolates were positive for both floR and cmlA.As these antimicrobial agents may potentially inducecross-resistance between animal and human bacterialpathogens, their prudent use in veterinary medicine ishighly recommended.

Dissemination and tracking of Salmonella spp. in integrated broiler operation

Controlling Salmonella in integrated broiler operation is complicated because there are numerous potential sources of Salmonella contamination, including chicks, feed, rodents, wild poultry operations, and the processing plant. The objective of this study was to investigate the distribution of Salmonella through all phases of two integrated broiler operations and to determine the key areas related to the control of all known sources of infection. Two different Salmonella serotypes were observed at integrated broiler chicken company A. S. enteritidis, the predominant company A isolate, was consistently found in the breeder farm, hatcheries, broiler farms, and chicken slaughterhouse. At company B, a total of six different serotypes, S. heidelberg, S. senftenberg, S. enteritidis, S. blockley, S. gallinarum, and S. virchow, were detected. Although S. heidelberg was not found in the broiler farms, it was consistently found in the breeder farm, hatcheries, and chicken slaughterhouse. In addition, S. enteritidis was found in the hatcheries, broiler farm, and chicken slaughterhouse. In order to obtain the genetic clonality, 22 S. enteritidis isolates were digested with XbaI and analyzed by pulsed-field gel electrohporesis (PFGE). A difference in the PFGE pattern was found to be related to the origin of the integrated broiler operation. These data support the critical need to control Salmonella in breeder farms and hatcheries, and demonstrate important points related to the control of infection in large-scale poultry operations of Korea.

Macrorhabdus ornithogaster in ostrich, rhea, canary, zebra finch, free range chicken, turkey, guinea-fowl, columbina pigeon, toucan, chuckar partridge and experimental infection in chicken, japanese quail and mice

Since 2000, Macrorhabdus ornithogaster "megabacteriosis" has been diagnosed in the avian diseases laboratory in a diversity of avian species and varied spectrum of disease. The disease in some species (chickens, turkeys, guinea fowls) was clinically characterized by emaciation, prostration, loss of appetite, cachexia and death, with a typically chronic course. A more acute disease was observed in finches (canary-Serinus and zebra-Taeniopygia) and budgerigars (Melopsittacus undulatus). The large rod shaped organism, visible from 100 times magnification, with and without staining, could be detected in sick and also in reasonably normal individuals of some species, such as chickens, turkeys, quails and pigeons. In rheas (Rhea americana), ostriches (Struthio camelus), canaries, zebra-finches, guinea-fowl (Numida meleagris) and budgerigars. The disease was severe, causing to up to 100 percent mortality. The infection could be detected in some species along with other infectious or disease problems, such as endoparasites (helminths, coccidia) and ectoparasitism (order Mallophaga or/and order Acarina). The cultivation of M. ornithogaster was successfully achieved in solid and liquid media, originated from chickens (four isolates), guinea fowl (1 isolate), chuckar partridge (1 isolate) and canary (1 isolate). A very interesting finding at microscopy was motility of M. ornithogaster, as detected both in cultures obtained on agar for pathogenic fungi and passaged into thioglycolate broth, as well as on samples observed in wet preparations from in vivo. Differences in colony aspects were noted among the isolates. Experimental infections were attempted in chicken and japanese quail, using a chicken isolate, allowing the detection of the organism in the proventriculus and liver in apparently normal birds. One chicken isolate was injected intraperitoneally in Balb/c mice and resulted in 100 percent mortality.

Desde 2000, diversos casos de infecção e doença por Macrorhabdus ornithogaster (megabacteria) foram diagnosticados no Setor de Doenças das Aves (Escola de Veterinária da UFMG). A doença clínica foi caracterizada por emagrecimento, prostração, perda do apetite, caquexia e morte, em curso crônico, embora com forma mais aguda em canários e periquitos. O microrganismo grande, em forma de bastão, visível a partir de 100 aumentos sem e com coloração, pode também ser detectado em aves de aspecto clínico normal, principalmente galinhas, perus, codornas e pombos. Em emas (Rhea), avestruzes (Struthio camelus), canários, mandarins, galinhas da Angola (Numida meleagris) e periquitos Australianos (Melopsittacus undulatus), a severidade da doença foi sempre maior, ocasionando até 100 por cento de mortalidade em alguns plantéis. Na maioria das espécies a doença foi detectada em aves com endo e/ou ectoparasitismo. O cultivo de M. ornithogaster foi obtido em meio sólido (ágar para fungos patogênicos) e subcultivado em meio líquido (thioglicolato), do proventriculo de galinha, galinha da Angola, perdiz de chuckar e canário. O resultado mais surpreendente na microscopia de M. ornithogaster foi a presença de motilidade, detectada tanto de cultivos in vitro quanto de preparações úmidas de in vivo. Diferenças nos aspectos das colônias foram notadas entre os isolados. Infecções experimentais em galinha (SPF) e codorna japonesa permitiram a detecção do organismo nos proventrículos das aves de aspecto normal. Nas codornas, à necropsia notaram-se hemorragias hepáticas. A infecção experimental em camundongos via intraperitoneal resultou em 100 por cento de mortalidade, também com lesões hepáticas. Aspectos do cultivo, a importância da doença, as espécies de aves susceptíveis e seu papel na epidemiologia são discutidos.

Analysis of Salmonella enterica serotype Enteritidis isolated from human and chickens by repetitive sequence-PCR fingerprinting, antibiotic resistance and plasmid profiles

A total of 22 Salmonella enterica serotype Enteritidis (S. Enteritidis) strains isolated from human and chicken were subjected to DNA fingerprinting by repetitive sequence PCR using ERIC and BOX primers, antibiotic resistance and plasmid patterns. Both ERIC and BOX PCR amplification data revealed a highly genetic homogeneity between isolates from human and chicken except one isolate, which originated from chicken and showed a different DNA band pattern from others. Eleven of 22 S. Enteritidis isolates (50%) were resistant to more than one antibiotics and characterized by 5 resistance patterns. The most common pattern was penicillin resistant (63.6%). Only one isolate from chicken showed a multiple drug resistance patterns to 4 antibiotics. All 22 S. Enteritidis isolates harbored more than two plasmids with eight different plasmid profiles including two to six plasmids with approximate molecular size ranging from 1.9 to 21 kb. A band of 15 kb size was detected in all isolates tested, however, the band sizes smaller than 15 kb were found only in isolates from chicken.

Occurrence of Campylobacter species in poultry forms in Lagos area of Nigeria.

Samples of poultry droppings were collected from fifty poultry farms in Lagos area of Nigeria. Campylobacter species were isolated from serially diluted samples using Bolton selective enrichment medium under microaerophilic condition at 42 degrees C. Samples of water, both treated and untreated used in the farms were also analysed in the same manner. Fifteen Campylobacter strains were isolated from the droppings. All the strains were identified using biochemical characteristics differentiating Campylobacter species. They were identified as hippurate positive (C. jejuni) and negative species (C. coli). They were biotype' using the extended scheme described by Lior (1984). Campylobacter coli biotype 'I was the most prevalent (66.66%) followed by C. jejuni biotype 1 (20.00%). The biotype ii of the two species accounted for 6.67% each. The C. jejuni biotype III and IV were not isolated during the study. The preponderance of C. coli indicates that poultry can be a major reservoir for the spread of human campylobacteriosis. Poultry farmers, therefore, must ensure prompt and adequate means of disposal of waste materials as well as maintaining good sanitary conditions.

Characterisation of fowl adenoviruses from chickens affected with infectious hydropericardium during 1994-1998 in India.

In the present study characterisation has been done for six group I fowl adenoviruses (FAV) isolated from outbreaks of infectious hydropericardium (IHP) of chickens that occurred in different states/regions of India during the years 1994-98. These six viruses were identified as FAV serotype 4 by virus neutralisation and restriction endonuclease analyses. Antigenic analyses of the viruses revealed close relationship (R-values 0.93-0.96). Under the experimental conditions, we have been able to induce IHP using FAV serotype 4 isolate AD: 411 and were also able detect FAV antigens in myocardial tissues by immunofluorescence assay (a new observation), an indication that IHP causing FAV serotype 4 strain replicate in myocardial tissue. Restriction endonuclease analysis of the viral genomes (approximately 46 Kb), using Hind III, Sma I, Xba I, Bam HI, Pst I and Dra I produced identical genetic profiles. Pst I and Bam HI profiles for these six vitus isolates were identical to those published earlier for an IHP causing Pakistani FAV serotype 4 isolate KR31. The identical genetic profiles of viruses, chronology of the outbreaks of IHP in Pakistan during 1989 onward and later in Jammu and Kashmir, India (1994), suggest that FAV serotype 4 isolates involved in outbreaks of IHP in India had probably spread from Pakistan. In order to prevent further spread and economic losses due to IHP in India, based on the antigenic relatedness data in this paper, any one of the six studied FAV serotype 4 isolates can be used as a candidate for mass production of CEH culture based killed vaccine.

Freqüência e patologia das infecçöes causadas por nematóides e cestóides em galinhas-d'angola (Numida meleagris Linnaeus, 1758) criadas extensivamente no estado do Rio de Janeiro, Brasil / Frequency and pathology of infeccions caused by nematodes and cestodes in free ranged guinea fowls (Numida meleagris Linnaeus, 1758) in the state of Rio de Janeiro, Brazil

Foi realizada uma pesquisa sobre as helmintoses que ocorrem em galinhas-d'angola criadas extensivamente no estado do Rio de Janeiro, através de exames clínicos, necropsias e exames histopatológicos de 36 aves. Os nematóides identificados com as respectivas prevalências e intensidades médias de infecçäo foram: Heterakis gallinarum 100,0 por cento e 194,08; Ascaridia numidae 75,0 por cento e 48,80; Capillaria spp. 52,8 por cento e 30,08; Dispharynx nasuta 44,4 por cento e 39,50; Eucoleus perforans 5,5 por cento e 0,55; Oxyspirura mansoni 2,8 por cento e 0,08. Raillietina (Paroniella) magninumida foi o único cestóide identificado, com prevalência de 47,2 por cento e intensidade média de 20,33. A. numidae, D. nasuta e E. perforans foram considerados os helmintos mais patogênicos. O nematóide Dispharynx nasuta foi registrado pela primeira vez no Brasil, neste hospedeiro.

Soroprevalência do Toxoplasma gondii em galinhas (Gallus gallus domesticus) de criaçöes domésticas, oriundas de propriedades rurais do norte do Paraná, Brasil / Seroprevalence of Toxoplasma gondii in back yard chicken (Gallus gallus domesticus) from rural properties in north region of Paraná state, Brazil

O Toxoplasma gondii é um protozoário que pode infectar todos os animais homeotérmicos. No presente trabalho, verificou-se a ocorrência de anticorpos anti-T. gondii em galinhas de criaçöes domésticas (fundo de quintal), oriundas de propriedades rurais localizadas no município de Jaguapitä, Estado do Paraná, Brasil. Estudos prévios demonstraram que o parasita está amplamente disseminado nas diferentes espécies de animais domésticos, de companhia e no homem nessas mesmas propriedades. Os soros foram submetidos à reaçäo de Imunofluorescência Indireta, sendo a positividade considerada para àqueles títulos maiores ou iguais a l6. Foram estudados 155 soros, sendo que 16 (l0,3 por cento) foram reagentes à toxoplasmose, e 139 (89,7 por cento) näo reagentes. Os títulos mais frequentes foram de 64 (5/31,2 por cento), l6 e 1024 (ambos com 4/25 por cento) e o maior título encontrado foi de 1024 (4/25 por cento). A soroprevalência do T. gondii näo foi influenciada pelo sexo da ave (Exato de Fisher p = 0,12), pela finalidade da criaçäo (corte ou postura, Exato de Fisher p = 0,39), pela percentagem de felinos sororeagentes na propriedade (r = -0,19 p = 0,27), nem pelo número de felinos sororeagentes (r = -0,09 p = 0,60). Através do presente trabalho, demonstrou-se uma ocorrência elevada da resposta sorológica ao T. gondii em aves domésticas oriundas de propriedades rurais neste município, conseqüentemente esses animais podem representar risco de infecçäo para o homem e para outros animais quando consumidos crus ou mal cozidos.

Study of Cyclospora cayetanensis in health care facilities, sewage water and green leafy vegetables in Nepal.

Cyclospora cayetanensis, a newly emerging parasite, is endemic in Nepal. A total of 2,123 stool specimens were collected from 3 health care facilities based on clinical symptoms during the period between 1995 to October, 1998. Out of these specimens, cayetanensis oocysts were found in 632 (29.8%). To identify possible sources for Cyclospora infection, drinking water, sewage water, green-leafy vegetables including fecal samples of various animals were collected and examined. The vegetable leaves were washed in distilled water then the washings, sewage water and drinking water were centrifuged and the sediment were examined microscopically. As a result, oocyst of Cyclospora were identified in sewage water and vegetable washings on four different occasions in June, August, October and November. The positive results were also confirmed as C. cayetanensis by development of 2 sporocysts after 2 week incubation period in potassium dichromate. A survey of 196 domestic animals from the same areas demonstrated that two chickens were positive for Cyclospora-like organism and others were negative. Although further studies are needed to clarify the direct link between Cyclospora infection and these sources, the results suggest that sewage water, green leafy vegetables are possible sources of infection and chickens could be possible reservoir host of Cyclospora in Nepal.