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This study describes the effect of bactofugation (10,000 × g in a continuous flow of 10,000 L/h) of three batches of raw milk on total bacterial count (TBC) and aerobic spore count, and it also shows the effect on the microbial diversity of spore-forming bacteria by analysing their genetic variability through molecular approaches. Given that milk must be preheated to approximately 55 °C before being bactofuged, for comparison, the three batches were evaluated at different stages as refrigerated raw, preheated, and bactofuged milk. For preheated milk, it was found that bactofugation caused a significant reduction (p < 0.05) of 99.52% (from 4.5 × 106 to 2.1 × 104 CFU/mL) in the mean TBC and of 95% (from 333 to 17 CFU/mL) in the aerobic mesophilic spore count. Due to the effect of bactofugation on preheated milk, a reduction of 82% was observed in both TBC and aerobic spore count. With respect to diversity, 107 isolates from raw milk, prior to bactofugation, and 16 isolates from bactofuged milk were recovered and grouped into 40 and 8 clusters, respectively. The predominant species detected in raw and preheated milk were Bacillus toyonensis (63% - 20 clusters) and Lysinibacillus fusiformis (15% - 8 clusters). Proportionally, B. toyonensis (69% - 6 clusters) and L. fusiformis (25% - 1 cluster) were predominant in bactofuged milk. B. pumilus, L. varians, B. flexus, B. invictae, and B. megaterium, bacteria with a known milk spoilage potential, were isolated from milk prior to bactofugation, and they reduced to undetectable levels in bactofuged milk. Bactofugation of milk, therefore, reduces the TBC and aerobic spore count, with a significant effect in reducing the microbial diversity of spore-forming bacteria, proportional to their incidence in raw milk. Therefore, bactofugation can be an alternative to increase the shelf life and technological potential of milk. Esse estudo descreve o efeito da bactofugação (10,000 × g em fluxo contínuo de 10,000 L/h) de três lotes de leite cru nas contagens bacterianas totais (CBT) e de esporos aeróbios, verificando também o efeito sobre a diversidade microbiana dos formadores de esporos por abordagem molecular de variabilidade genética. Como o leite a ser bactofugado deve ser pré-aquecido (=55ºC), os três lotes foram avaliados enquanto cru refrigerado, pré-aquecido e bactofugado. Em associação com o pré-aquecimento, foi verificado que a bactofugação promoveu a redução significativa (p < 0.05) de 99,52% (4,5 x 106 para 2,1 x 104 UFC/mL) na contagem bacteriana total e 95% (333 para 17 UFC/mL) dos esporos aeróbios mesófilos. Considerando o efeito isolado da bactofugação sobre o leite já pré-aquecido, foi observada a redução de 82% tanto para CBT quanto para formadores de esporos. Em relação à diversidade, foram recuperados 107 isolados do leite anterior à bactofugação e 16 isolados do leite bactofugado, agrupados em 40 e 8 clusters, respectivamente. Foi observado predomínio das espécies Bacillus toyonensis (63% - 20 clusters) e Lysinibacillus fusiformis (15% - 8 clusters) no leite cru e pré-aquecido e, proporcionalmente, B. toyonensis (69% - 6 clusters) e L. fusiformis (25% - 1 cluster) no leite bactofugado. Bacillus pumilus, Lysinibacillus varians, B. flexus, B. invictae, e B. megaterium, isolados do leite antes da bactofugação e com potencial deteriorante conhecido, foram reduzidos a níveis indetectáveis no leite bactofugado. A bactofugação do leite, portanto, reduz a CBT e as contagens de esporos aeróbios, também com efeito significativo na redução da diversidade de formadores de esporos, proporcionalmente, conforme a sua incidência no leite cru, tendo potencial para ser utilizado como alternativa para aumento da vida útil e potencial tecnológico do leite.

Noroviruses (NoVs) are a leading cause of acute gastroenteritis (diarrhoea and vomiting) worldwide. The virus was reported from humans and later was identified in different symptomatic and/or asymptomatic animal species, such as bovine, ovine, swine, canine and feline. In animals NoV infection is supposedly associated with enteritis; however, the pathogenic role of NoV infection and its impact in animal health are currently uncertain. NoVs have a positive sense, single-stranded RNA genome of diverse molecular feature. Currently, NoVs are classified into six genogroups (GI-GVI), with the proposed seventh (GVII) and at least 31 genotypes. The GI, GV and GVI are species-specific genogroups, while the GII, GIII and GIV share different host species, including humans. The close genetic relationship among different NoV strains detected from both humans and animals raised the possibility of this virus has zoonotic potential. This review aimed to address the NoV classification, morphological and molecular features, epidemiology, pathogenesis and pathology, diagnosis, treatment and management. A broad approach is given on the importance of NoVs as a cause of disease in livestock and pets and on studies into the potential for zoonotic transmission and public health.

Helicobacter pylori infection causes gastritis and can induce gastric adenocarcinoma and MALT lymphoma in humans. The aim of this study was to determine in dogs whether there is an association between Helicobacter spp. infection in gastric mucosa, histological lesions, including epithelial cell proliferation and cell adhesion. Gastric biopsies of 12 dogs with gastric disturbances and 25 healthy dogs were evaluated. Warthin-Starry staining (WS) and PCR assay were performed to confirm the presence of helicobacteria. The Helicobacter species were determined by PCR assay with speciesspecific primers for H. heilmannii, H. bizzozeronii or H. salomonis, H. felis and H. pylori. Mucosal lesions were evaluated by haematoxylin and eosin (HE) and epithelial proliferation was determined by AgNOR and PCNA methods. Cell adhesion was evaluated by the expression of E-cadherin by epithelial cells. Helicobacter spp. was confirmed in 75.7% (28/37) and 73.0% (27/37) of the samples by WS and PCR, respectively. H. bizzozeronii was the species most frequently detected (37%); co-infection was observed in six (22%) dogs. Histological changes in the lamina propria included mild chronic gastritis, fibrosis, glandular degeneration, and presence of lymphoid aggregates. There was a significant association between H. heilmannii infection and the presence of lymphoid follicles (p < 0.046). In this study no significant change in epithelial proliferation or E-cadherin expression were detected in the gastric mucosa of dogs infected with Helicobacter spp.Infecção por Helicobacter pylori provoca gastrite e pode induzir adenocarcinoma gástrico e linfoma tipo MALT em seres humanos. O objetivo deste estudo foi determinar se existe associação entre infecção por Helicobacter spp. na mucosa gástrica de cães e lesões histológicas, incluindo proliferação epitelial e adesão celular. Foram avaliadas biópsias de 12 cães com distúrbios gástricos e de 25 cães saudáveis. A coloração de Warthin-Starry (WS) e o método de PCR foram utilizados para confirmar a presença de helicobactérias. As espécies de Helicobacter foram determinadas por PCR com primers espécie-específicos para H. heilmannii, H. bizzozeronii ou H. salomonis, H. felis e H. pylori. As lesões da mucosa foram avaliadas por hematoxilina e eosina (HE) e a proliferação epitelial foi determinada pelos métodos AgNOR e PCNA. A adesão celular foi avaliada por meio da expressão de E-caderina nas células epiteliais. Helicobacter spp. foi confirmado em 75,7% (28/37) e 73,0% (27/37) das amostras por WS e PCR, respectivamente. H. bizzozeronii foi a espécie mais frequentemente detectada (37%); e coinfecção foi observada em seis (22%) cães. As alterações histológicas na lâmina própria foram gastrite crônica discreta, fibrose, degeneração glandular e presença de agregados linfóides. Houve associação significativa entre a infecção por H. heilmannii e a presença de folículos linfóides (p

GISAID EpiFlu database accession numbers for EIV segments. (DOCX 43Â kb)

Trinta e cinco vacas de rebanhos leiteiros da região Norte do estado do Paraná, com histórico de abortamento, foram pesquisadas sorologicamente para verificar a presença de anticorpos contra Neospora caninum, Toxoplasma gondii, Leptospira spp., Brucella abortus, BHV-1 e BVDV. Vinte e uma vacas apresentaram títulos sorológicos compatíveis com infecção. Todas elas, soropositivas para N. caninum, foram também soropositivas para outros agentes infecciosos, sugerindo a possibilidade de associação desses agentes nos problemas reprodutivos de bovinos, no estado do Paraná.

Helicobacter pylori is the most common gastric bacteria of human beings. Animal-borne helicobacter have been associated with gastritis, ulceration, and gastric mucosa-associated lymphoid-tissue lymphoma in people. We attempted to identify the species of Helicobacter spp. that infect human beings in north Paraná, Brazil. Samples of gastric mucosa from 38 dyspeptic patients were analyzed by optic microscopy on silver stained slides, polimerase chain reaction (PCR), and enzymatic cleavage. Genus and species-specific primers to H. pylori, H. heilmannii, H. felis, and consensual primers to H. bizzozeronii or H. salomonis were used. The PCR products were submitted to enzymatic cleavage by VspI (Helicobacter spp. product) and HinfI (species products) enzymes. Thirty-two out of 38 patients evaluated had 3.2 to 5 µm long bacteria that resembled H. pylori in Warthin-Starry stained slides and were positive to the genus Helicobacter by PCR. In 30 of these patients the bacteria were identified as H. pylori. Two samples positive by silver stain were negative to all species tested by PCR. None of the 38 samples was positive to animal-origin helicobacter species. These results show that PCR and enzymatic restriction are practical methods to identify the species of helicobacters present in gastric mucosa of human beings. People in north Paraná appear to be infected mostly with H. pylori.