Caracterização da microbiota de carcaças de frango de corte em matadouros frigoríficos através do sequenciamento de alto rendimento

Analisar e entender o perfil da comunidade bacteriana das carcaças de frango de corte durante o processo de abate pode ser uma ferramenta útil para a agroindústria avícola produzir alimentos com maior inocuidade. O microbioma do matadouro-frigorífico de frangos de corte tem um grande impacto na segurança do produto final gerado. O manuseio e o consumo de produtos de origem aviária têm sido associados à transmissão de importantes patógenos aos consumidores, como Salmonella spp., Campylobacter jejuni, Escherichia coli e Listeria monocytogenes. Porém outras bactérias patogênicas também podem estar relacionadas com a cadeia avícola. Nos matadouros-frigoríficos, a contaminação da carcaça de frango por bactérias patogênicas pode ocorrer durante todas as etapas do processamento, incluindo abate, sangramento, evisceração, lavagem e refrigeração. Este trabalho tem como objetivo realizar a técnica de sequenciamento de próxima geração de alto rendimento (HT-NGS) para identificar o microbioma presente nas carcaças de frango de corte durante o processo de abate em matadouros-frigoríficos. Foram coletadas 312 carcaças de frango de corte em três matadouros-frigoríficos do Rio Grande do Sul com Serviço de Inspeção Federal (SIF). Também foram coletados 200 mL de água do chiller em três pontos distintos (início, meio e fim) do tanque em cada matadouro-frigorífico sendo realizado um pool para cada ponto amostrado. O pool correspondente a cada ponto foi constituído de 100 mL, sendo deste total 50 mL por ponto amostrado de cada visita. A comunidade microbiana das carcaças de frango foi dominada pelo Reino Bacteria em todos os matadouros-frigoríficos. Foram identificados quatro filos, 11 classes, 15 ordens e 22 famílias. No matadouro-frigorífico A foram identificados 20 gêneros e nos matadouros-frigoríficos B e C foram identificados 28 gêneros em cada. No matadouro-frigorífico A foram identificadas 26 espécies e nos matadouros-frigoríficos B e C foram identificadas 37 espécies em cada. No chiller foram identificadas 17 espécies e não foi observada nenhuma espécie comum aos três estabelecimentos avícolas. Na área limpa (embalagem final) foram identificadas 21 espécies, sendo a espécie C. jejuni a única comum aos três estabelecimentos avícolas. O processo de abate reduziu o número de espécies e a quantidade de sequências identificadas dos microrganismos. O chiller não foi capaz de eliminar ou diminuir a quantidade de alguns contaminantes detectados, independentemente do matadouro-frigorífico avaliado. Entre os microrganismos com potencial patogênico detectado, foi observada a presença do C. jejuni nos três matadouros-frigoríficos. O microbioma das carcaças de frango de corte foi diferente para todos os matadouros-frigoríficos. Os resultados demonstram que o microbioma associado à carcaça varia conforme o matadouro-frigorífico, bem como de acordo com a origem do lote.Analyzing and understanding the profile of the bacterial community of broiler chicken carcasses during the slaughter process can be an useful tool for the poultry industry to produce microbiologically safer food. The microbiome of a broiler slaughterhouse has a great impact on the safety of the final product generated. On the other hand, the handling and consumption of products of avian origin have been associated with the transmission of important pathogens to consumers, such as Salmonella spp., Campylobacter jejuni, Escherichia coli and Listeria monocytogenes. However, other pathogenic bacteria may also be related to the poultry chain. In slaughterhouses, chicken carcass contamination by pathogenic bacteria can occur during all stages of processing, including slaughter, bleeding, evisceration, washing and refrigeration. This work aims at performing the next generation high throughput sequencing technique (HT-NGS) to identify the microbiome present in broiler carcasses during the slaughter process in slaughterhouses. A total of 312 broiler carcasses were collected from three slaughterhouses in the State of Rio Grande do Sul, Brazil, all served with Federal Inspection Service (SIF). 200 mL of water from the chiller were also collected at three different points (beginning, middle and end) of the chiller tank in each processing plant, with a pool being made for each point sampled. The pool corresponding to each point consisted of 100 mL, of which 50 mL per point was sampled at each visit. The microbial community of chicken carcasses was dominated by the Bacteria Kingdom in all slaughterhouses. Four phyla, 11 classes, 15 orders and 22 families were identified. In slaughterhouse A, 20 genera were identified and in slaughterhouses B and C, 28 genera were identified in each. In slaughterhouse A, 26 species were identified and in slaughterhouses B and C, 37 species were identified in each. In the chiller, 17 species were identified and no species common to the three poultry establishments were observed. In the clean area (final packaging), 21 species were identified, with the species C. jejuni being the only one common to the three poultry establishments. The slaughter process reduced the number of species and the amount of identified sequences of microorganisms. The chiller was not able to eliminate or reduce the amount of some contaminants detected, regardless of the slaughterhouse evaluated. As referred, among the microorganisms with pathogenic potential detected, the presence of C. jejuni was observed in all three slaughterhouses. The microbiome of broiler carcasses was different for all slaughterhouses. The results demonstrate that the microbiome associated with the carcass can vary according to the slaughterhouse sampled, as well as according to the origin of the batch being processed

Bovine herpesviruses do not play a major role in the differential diagnosis of rabies in cattle in Southern Brazil

Background: Rabies has long been recognized as the major cause of encephalitis in cattle in Latin American countries. It has been estimated that nearly 50.000 cattle heads per year are lost due to encephalitis in that subcontinent, with a signifi cant economic impact on cattle productive chains. In Brazil only, 2.500 to 3.000 cattle heads are estimated to be lost every year due to rabies. However, it is believed that rabies incidence in cattle is much larger, since usually only a few samples from affected animals in disease outbreaks are submitted to diagnostic laboratories. Rabies encephalitis is promptly and accurately diagnosed; however, particularly when rabies is excluded as causa mortis, the agent responsible for neurological disease of infectious origin often remains undetermined. Two bovine herpesviruses (BoHVs), bovine herpesvirus type 1 (BoHV-1) and bovine herpesvirus type 5 (BoHV-5) are major pathogens of cattle which are widely disseminated in Brazil. As usual in herpesvirus’ biology, these tend to infect a large number of hosts and establish lifelong latent infections which may occasionally be reactivated. Both viruses, particularly BoHV-5, are often recovered from cases of neurological disease in cattle. The participation of BoHVs in the differential diagnosis of rabies must be evaluated. Besides, there might be associations between the occurrence of rabies and BoHV infections that deserve investigation. The aim of this study was to investigate whether bovine herpesvirus 1 and 5 would play a signifi cant role in cases of neurological disease where rabies was the presumptive clinical diagnosis. In addition, associations between the occurrence of rabies and BoHV infections were searched for. The approach adopted for conducting such investigations was based on the search for viral nucleic acids as well as classical virus isolation on tissues of cattle submitted to rabies diagnosis over a two-year Materials, Methods & Results: Brain tissue samples of 101 cattle originally submitted to rabies diagnosis were collected over a two year period (2009-2010) from various municipalities within the state of Rio Grande do Sul, Brazil. Thirty nine of these samples had the diagnosis of rabies confi rmed by standard laboratory diagnostic methods. Aliquots of tissues were submitted to DNA extraction and examined in search for genomes of bovine herpesviruses (BoHV) types 1 (BoHV-1) and 5 (BoHV-5) by as well as for infectious virus. Bovine herpesvirus genomes were detected in 78/101 (77.2%) samples, in which BoHV-1 genomes were detected in 26/78 (25.7%), BoHV-5 genomes in 22/78 (21.8%) and mixed BoHV infections (BoHV-1 and BoHV-5 genomes) were detected in 30/101 (29.7%) samples. In the 39 samples with confi rmed rabies diagnosis, BoHV-1 DNA was detected in 9/39 (23%), BoHV-5 DNA in 6/39 (15.4%) and mixed infections with both BoHV types in 16/39 (41%) samples. However, no infectious herpesvirus was recovered from any of the specimens examined. Discussion: The high prevalence of BoHV1 and BoHV-5 infections was evidenced in the sampled population, but the absence of infectious BoHVs indicate that these were not associated to the occurrence of the cases of encephalitis where rabies was the primary suspicion. In addition, no association was detected between occurrence of rabies and detection of BoHVs, since the frequency of detection of herpesvirus genomes did not signifi cantly differ between rabies-positive and rabies-negative samples. The detection of BoHV DNA in scattered areas of the brain with no infectious virus suggests that latency may take place in different regions of the brain

Detecção de dna de herpesvírus bovino em encéfalos de bovinos submetidos ao diagnóstico de raiva

Os herpesvírus bovino tipo 1 (BoHV-1) e 5 (BoHV-5) são alfaherpesvírus freqüentemente associados a meningoencefalites. Por outro lado, o vírus da raiva é o agente mais frequentemente identificado como causador de encefalites virais em bovinos no Brasil. O objetivo do presente estudo foi examinar a ocorrência de infecções por BoHV-1 e/ou BoHV-5 em amostras de tecido encefálico bovino submetidas ao diagnóstico de raiva e avaliar seu possível envolvimento nos quadros de encefalite que originaram a suspeita de raiva. Para tanto, 101 amostras desses tecidos, sendo 39 positivas para raiva e 62 negativas, recebidas pelo órgão oficial responsável pelo diagnóstico de raiva no Estado do Rio Grande do Sul (IPVDF) no período de 2009- 2010, foram submetidas a exames buscando o isolamento viral e amplificação de genomas de herpesvírus bovinos. Ao isolamento viral, todas as amostras foram negativas para vírus infeccioso após a realização de três passagens cegas em células MDBK. As mesmas foram submetidas à amplificação por “nested PCR” (nPCR) para a pesquisa de genomas de BoHV-1 e BoHV-5. Das 101 amostras totais analisadas esta técnica revelou que 25,7% (26/101) continham genomas de BoHV-1 e 21,8% (22/101) continham genomas de BoHV-5. Genomas de ambos os tipos foram identificadas em 30 (29,7%) amostras. Entre as amostras que foram também positivas para raiva em 23% (9/39) foram detectados genomas de BoHV-1 e em 15,4% (6/39) continham genomas de BoHV-5. Em 16 destas 39 amostras (41%) foram detectados genomas de BoHV-1 e BoHV-5. Em contrapartida, nas amostras negativas para o vírus rábico, 27,4% (17/62) também foram positivas para BoHV-1, 25,8% (16/62) foram positivas para BoHV-5. Detectaram-se os genomas de ambos BoHVs em 22,6% (14/62) dos animais. Estas diferenças não foram estatisticamente significativas, indicando não haver correlação entre a ocorrência de raiva e infecções por herpesvírus na amostragem realizada. Estes resultados indicam que, embora as infecções por BoHV-1 e BoHV-5 tenham apresentado elevada incidência nessas amostras, não havia vírus infeccioso nas mesmas, sugerindo infecções latentes sem envolvimento aparente nos quadros de encefalite que originaram a suspeita inicial de raiva.Bovine herpesvirus type 1 (BoHV-1) and 5 (BoHV-5) are alphaherpesviruses associated with a number of clinical manifestations in cattle, including encephalitis. On the other hand, rabies virus is the agent most frequently identified as cause of viral encephalitis in cattle in Brazil. The aim of this study was to examine the occurrence of BoHV-1 and / or BoHV-5 in bovine brain tissue samples submitted to rabies diagnosis. The search was carried out by virus isolation and nested polymerase chain reaction (PCR) in brain tissues of cattle submitted to rabies diagnosis in the state of Rio Grande do Sul in the period 2009-2010. One hundred and one brain samples from cattle with signs of neurological disease, of which 39 were positive and 62 negative for rabies, were used in this study. At virus isolation, all samples were negative for the presence of infectious herpesviruses after three successive passages in MDBK cells. Of the 101 total samples analyzed, this test revealed that 25.7% (26/101) of cattle were infected with BoHV-1 and 21.8% (22/101) were infected with BoHV-5. Genomes of both types were detected in 29.7% (30/101) samples. With the 39 samples positive for rabies virus, BoHV-1 genome was detected in 23% (9/39) and 15.4% (6/39) were positive for BoHV-5 as well as in 41% (16/39) of these samples, which were positive for both BoHVs. On the other hand, the negative samples for rabies virus, 27.4% (17/62) also were positive for BoHV-1, as well and 25.8% (16/62) were positive for BoHV-5. Genomes of both BoHVs were detected in 22.6% (14/62) of the specimens. These differences were not statistically significant indicating no correlation between the occurrence of rabies and herpesvirus infections in the animals. These results do not imply that the herpesviruses detected, even showing a high incidence, were the causative agents of meningoencephalitis in the samples tested, once it was not possible to isolate virus in its infectious form, however it suggests a latent infection in the animals involved with neurological signs of meningoencephalitis whose primary suspicion was rabies

Laparoscopic-assisted treatment of pyometra associated with mammary fibroadenomatous hyperplasia in a cat

This paper describes a case of laparoscopic-assisted ovariohysterectomy in a female cat presenting pyometra and mammary fibroadenomatous hyperplasia. Using four portals, mesovarium were ligated by titanium ligature clips whereas the uterine vessels were occluded by video-assisted conventional ligatures. There were no postoperative complications. Video-assisted technique can be an alternative method for treatment of pyometra and cystic endometrial hyperplasia in female cats

Ciência Rural, v.44, n.3, mar

ABSTRACT This paper describes a case of laparoscopic-assisted ovariohysterectomy in a femal

Antibiofilm activity of the biosurfactant and organic acids against foodborne pathogens at different temperatures, times of contact, and concentrations

Biofilm formation has been suggested to play a significant role in the survival of pathogens in food production. Interest in evaluating alternative products of natural origin for disinfectant use has increased. However, there is a lack of information regarding the effects of biosurfactants and organic acids on Salmonella enterica serotype Enteritidis, Escherichia coli, and Campylobacter jejuni biofilms, mainly considering temperatures found in environments of poultry processing, as well as simulating the contact times used for disinfection. The aim of this study was to evaluate the antibiofilm activity of rhamnolipid, malic acid, and citric acid on the adhesion of S. Enteritidis, E. coli, and C. jejuni on polystyrene surfaces at different temperatures (4, 12, and 25 °C), compound concentrations, and times of contact (5 and 10 min), and to analyze the potential use of these compounds to disrupt formed biofilms. All three compounds exhibited antibiofilm activity under all analyzed conditions, both in the prevention and removal of formed biofilms. Contact time was less important than temperature and concentration. The antibiofilm activity of the compounds also varied according to the pathogens involved. In the food industry, compound selection must consider the temperature found in each stage of product processing and the target pathogens to be controlled