Implementation of a milking system and quality management for buffalo milk at an experimental station in Rio Grande do Sul, Brazil / Implementação de um sistema de ordenha e manejo da qualidade do leite bubalino em uma estação experimental no Rio Grande do Sul, Brasil

Buffalo milk production is an activity that has grown in recent years, mainly due to the physicochemical characteristics found in this type of milk, which is the second most produced in the world. The objective of this study was to implement a milking system and monitor the identity and quality standards of buffalo milk produced at the Experimental Agronomic Station of the Federal University of Rio Grande do Sul. Samples were collected fortnightly between June and November 2021, totaling ten collections. The means obtained at the end of the monitoring were 4.84 g/100 g for fat, 4.64 g/100 g for protein, 5.06 g/100 g for lactose, 15.26 g/100 g for total solids, 10.42 g/100g for non-fat solids, 0.18 g lactic acid/100 mL of milk for acidity, 1.037 g/cm3 for density, -0.533°C for cryoscopic index, 3.5 x 105 cells/mL for somatic cell count, and 8.0 x103 CFU/ml for standard plate count. Residues of antibiotics and antiparasitics were not found. An increase in the concentration and frequency of some fatty acids was observed, after an increment in the nutritional management of the animals, in addition to predominance of palmitic acid (C16:0) and oleic acid (C18:1- cis (n9)). The results indicate a significant improvement in the quality of the milk in the assessment period, due to the corrective actions that were established, and the standard plate count showed good agricultural practices as to milk sourcing and the hygiene of the materials, being below the limits required by NI 76. The buffalo milk produced by the herd at the Experimental Station showed physicochemical and microbiological results in accordance with the legislation, being a raw material to be explored in the prospection of derivative products.

A produção de leite bubalino é uma atividade que tem crescido nos últimos anos, principalmente devido às características físico-químicas apresentadas pelo leite, que é o segundo mais produzido no mundo. O objetivo deste trabalho foi implementar um sistema de ordenha e monitorar os padrões de identidade e qualidade do leite bubalino produzido na Estação Experimental Agronômica da Universidade Federal do Rio Grande do Sul. Amostras foram coletadas quinzenalmente entre os meses de junho e novembro de 2021, totalizando dez coletas. As médias obtidas ao final do monitoramento foram de 4,84 g/100 g para gordura, 4,64 g/100 g para proteína, 5,06 g/100g para lactose, 15,26 g/100 g sólidos totais, 10,42 g/100g para sólidos não gordurosos, 0,18 g ácido lático/100 mL de leite para acidez, 1,037 g/cm3 para densidade, -0.533°C para índice crioscópico, 3.5 x105 cél/mL para contagem de células somáticas e 8.0 x103 UFC/ml para contagem padrão em placa. Não foram identificados resíduos de antibióticos e antiparasitários. Observou-se aumento de concentração e frequência de alguns ácidos graxos, após incremento de manejo nutricional dos animais, além da predominância dos ácidos palmítico (C16:0) e oleico (C18:1- cis (n9). Os resultados indicam que houve uma melhora significativa na qualidade do leite no período avaliado, decorrente das ações corretivas que foram estabelecidas, e a contagem padrão em placa demonstrou boas práticas agropecuárias na obtenção do leite e na higienização dos materiais, estando abaixo dos limites exigidos pela IN 76. O leite de búfala produzido pelo rebanho da Estação Experimental apresentou resultados físico-químicos e microbiológicos de acordo com a legislação, sendo uma matéria-prima a ser explorada na prospecção de produtos derivados.

Influence of the food matrix on dual-species biofilm of Listeria monocytogenes and Pseudomonas fluorescens

In the food industry, the formation of biofilms results in serious microbial recontamination problem. This work aimed to study Listeria sp. obtained from sliced products and food handling surfaces, for the preparation of food of animal origin to be consumed cold. Listeria monocytogenes, Listeria innocua and Listeria seeligeri were detected. In the evaluation of antibiotic susceptibility, the isolates were sensitive to ampicillin and penicillin, which were the main antibiotics used in treatment. The isolates showed the ability to form biofilm in microplate, in stainless steel and polypropylene surfaces, with a variation of sessile cells between 4.29±0.23 and 7.03±0.01 log10 CFU cm-2 in TSBYE. This ability was also observed in food matrix composed of UHT whole milk in mono-species cultivation and in associated cultivation of L. monocytogenes with Pseudomonas fluorescens. The application of sanitizers peracetic acid and sodium hypochlorite revealed efficiency in the eradication of adhered cells and biofilms formed in stainless steel surfaces. Therefore, Listeria sp. showed to be persistent and able to form biofilm in mono-species cultivation or associated with P. fluorescens, under different conditions. Taking these aspects into account, the need for proper hygiene in the food production process is highlighted in order to avoid risks to the consumers health.(AU)

Microbial contamination and antimicrobial resistance profiles indicate potential risks of infection at the Veterinary Medical Teaching Hospital - UFRGS, Porto Alegre, Brazil

Background: This study aimed to assess the level of bacterial contamination in the Small Animals Sector of the VeterinaryMedical Teaching Hospital (HCV) of the Universidade Federal do Rio Grande do Sul (UFRGS). Firstly, a committee wasinvited to complete a questionnaire and to list critical sample sites for collection. With the identification of the places to besampled, collections were made with sterile swabs on different surfaces of environments of the HCV. The identification ofimportant bacteria in the veterinary area, in the different sampled environments, raises the concern for hygiene proceduresin the veterinary hospital environment.Materials, Methods & Results: Sixteen samples were collected from these different areas, and microbiological analyses wereperformed. Standard counts of viable and strictly aerobic mesophilic microorganisms were realized. Collections were madeto assess ambient air quality. With the microbiological analysis performed, bacteria of clinical importance were identified.To assess the resistance profile of the bacteria, the susceptibility test to antimicrobials was performed. MALDI-TOF/MSmeasurement identified 29 bacteria at the genus level and 10 bacteria at the species level and the antimicrobial susceptibility test was realized. Most of the isolates identified (60%) were bacteria of the genus Staphylococcus spp. Regardingantimicrobial susceptibility analysis the 10 bacteria identified at the species level were assessed. Test results showed thatthe isolates S. aureus, S. epidermidis and S. haemolyticus - collected from treatment room 2 - and S. haemolyticus, whichhad been isolated from samples from treatment room 2 of the cattery, presented multiresistance. Pantoea ananatis isolatesfrom room 5 also showed a multiresistant profile for erythromycin, cephalothin, vancomycin and ampicillin. Micrococcusluteus isolates from the x-ray room and the kennel showed resistance to ceftazidime. Staphylococcus equorum isolates...

Microbial contamination and antimicrobial resistance profiles indicate potential risks of infection at the Veterinary Medical Teaching Hospital - UFRGS, Porto Alegre, Brazil

Background: This study aimed to assess the level of bacterial contamination in the Small Animals Sector of the VeterinaryMedical Teaching Hospital (HCV) of the Universidade Federal do Rio Grande do Sul (UFRGS). Firstly, a committee wasinvited to complete a questionnaire and to list critical sample sites for collection. With the identification of the places to besampled, collections were made with sterile swabs on different surfaces of environments of the HCV. The identification ofimportant bacteria in the veterinary area, in the different sampled environments, raises the concern for hygiene proceduresin the veterinary hospital environment.Materials, Methods & Results: Sixteen samples were collected from these different areas, and microbiological analyses wereperformed. Standard counts of viable and strictly aerobic mesophilic microorganisms were realized. Collections were madeto assess ambient air quality. With the microbiological analysis performed, bacteria of clinical importance were identified.To assess the resistance profile of the bacteria, the susceptibility test to antimicrobials was performed. MALDI-TOF/MSmeasurement identified 29 bacteria at the genus level and 10 bacteria at the species level and the antimicrobial susceptibility test was realized. Most of the isolates identified (60%) were bacteria of the genus Staphylococcus spp. Regardingantimicrobial susceptibility analysis the 10 bacteria identified at the species level were assessed. Test results showed thatthe isolates S. aureus, S. epidermidis and S. haemolyticus - collected from treatment room 2 - and S. haemolyticus, whichhad been isolated from samples from treatment room 2 of the cattery, presented multiresistance. Pantoea ananatis isolatesfrom room 5 also showed a multiresistant profile for erythromycin, cephalothin, vancomycin and ampicillin. Micrococcusluteus isolates from the x-ray room and the kennel showed resistance to ceftazidime. Staphylococcus equorum isolates...(AU)

Activity of the antimicrobial peptide P34 against bovine alphaherpesvirus type 1 / Atividade do peptídeo antimicrobiano P34 contra o alfaherpesvírus bovino tipo 1

Previous studies have demonstrated the antimicrobial activity of the peptide P34. In this study, the antiviral potential of P34 and the in vitro mechanism of action were investigated against bovine alphaherpesvirus type 1 (BoHV1). P34 exhibited low toxicity, a high selectivity index (22.9) and a percentage of inhibition of up to 100% in MDBK cells. Results from antiviral assays indicated that P34 did not interact with cell receptors, but it was able to inhibit the viral penetration immediately after pre-adsorption. In addition, BoHV1 growth curve in MDBK cells in the presence of P34 revealed a significant reduction in virus titer only 8h post-infection, also suggesting an important role at late stages of the replicative cycle. Virucidal effect was observed only in cytotoxic concentrations of the peptide. These findings showed that the antimicrobial peptide P34 may be considered as a potential novel inhibitor of in vitro herpesviruses and must encourage further investigation of its antiherpetic activity in animal models as well as against a wide spectrum of viruses.

A atividade antimicrobiana do peptídeo P34 já foi previamente demonstrada. Neste estudo, o potencial antiviral do P34 e o mecanismo de ação in vitro contra o alfaherpesvírus bovino tipo 1 (BoHV1) foram investigados. O P34 exibiu baixa toxicidade, alto índice de seletividade (22.9) e percentagem de inibição viral de até 100% em células MDBK. Os resultados dos ensaios antivirais indicaram que não interage com receptores celulares, mas é capaz de inibir a penetração viral, imediatamente após a pré-adsorção. Além disso, a curva de crescimento do BoHV1 em células MDBK na presença do P34 revelou uma significativa redução no título somente após 8h de infecção, sugerindo também uma importante atividade do peptídeo nas fases finais do ciclo replicativo. Efeito virucida frente / BoHV1 foi observado apenas em concentrações citotóxicas do peptídeo. Os dados obtidos indicam que o peptídeo antimicrobiano P34 pode ser considerado um potencial composto inibidor de herpesvírus, in vitro, e estimulam posteriores investigações sobre sua atividade anti-herpética em modelos animais, bem como contra outros vírus.

Activity of the antimicrobial peptide P34 against bovine alphaherpesvirus type 1 / Atividade do peptídeo antimicrobiano P34 contra o alfaherpesvírus bovino tipo 1

Previous studies have demonstrated the antimicrobial activity of the peptide P34. In this study, the antiviral potential of P34 and the in vitro mechanism of action were investigated against bovine alphaherpesvirus type 1 (BoHV1). P34 exhibited low toxicity, a high selectivity index (22.9) and a percentage of inhibition of up to 100% in MDBK cells. Results from antiviral assays indicated that P34 did not interact with cell receptors, but it was able to inhibit the viral penetration immediately after pre-adsorption. In addition, BoHV1 growth curve in MDBK cells in the presence of P34 revealed a significant reduction in virus titer only 8h post-infection, also suggesting an important role at late stages of the replicative cycle. Virucidal effect was observed only in cytotoxic concentrations of the peptide. These findings showed that the antimicrobial peptide P34 may be considered as a potential novel inhibitor of in vitro herpesviruses and must encourage further investigation of its antiherpetic activity in animal models as well as against a wide spectrum of viruses.(AU)

A atividade antimicrobiana do peptídeo P34 já foi previamente demonstrada. Neste estudo, o potencial antiviral do P34 e o mecanismo de ação in vitro contra o alfaherpesvírus bovino tipo 1 (BoHV1) foram investigados. O P34 exibiu baixa toxicidade, alto índice de seletividade (22.9) e percentagem de inibição viral de até 100% em células MDBK. Os resultados dos ensaios antivirais indicaram que não interage com receptores celulares, mas é capaz de inibir a penetração viral, imediatamente após a pré-adsorção. Além disso, a curva de crescimento do BoHV1 em células MDBK na presença do P34 revelou uma significativa redução no título somente após 8h de infecção, sugerindo também uma importante atividade do peptídeo nas fases finais do ciclo replicativo. Efeito virucida frente / BoHV1 foi observado apenas em concentrações citotóxicas do peptídeo. Os dados obtidos indicam que o peptídeo antimicrobiano P34 pode ser considerado um potencial composto inibidor de herpesvírus, in vitro, e estimulam posteriores investigações sobre sua atividade anti-herpética em modelos animais, bem como contra outros vírus.(AU)

Antiviral activity of a Bacillus sp: P34 peptide against pathogenic viruses of domestic animals

P34 is an antimicrobial peptide produced by a Bacillus sp. strain isolated from the intestinal contents of a fish in the Brazilian Amazon basin with reported antibacterial activity. The aim of this work was to evaluate the peptide P34 for its in vitro antiviral properties against canine adenovirus type 2 (CAV-2), canine coronavirus (CCoV), canine distemper virus (CDV), canine parvovirus type 2 (CPV-2), equine arteritis virus (EAV), equine influenza virus (EIV), feline calicivirus (FCV) and feline herpesvirus type 1 (FHV-1). The results showed that the peptide P34 exhibited antiviral activity against EAV and FHV-1. The peptide P34 inhibited the replication of EAV by 99.9% and FHV-1 by 94.4%. Virucidal activity was detected only against EAV. When P34 and EAV were incubated for 6 h at 37 °C the viral titer reduced from 10(4.5) TCID50 to 10(2.75) TCID50, showing a percent of inhibition of 98.6%. In conclusion, our results demonstrated that P34 inhibited EAV and FHV-1 replication in infected cell cultures and it showed virucidal activity against EAV. Since there is documented resistance to the current drugs used against herpesviruses and there is no treatment for equine viral arteritis, it is advisable to search for new antiviral compounds to overcome these infections.

Produção, purificação e caracterização de um peptídeo antimicrobiano produzido por uma linhagem de Bacillus sp. P34 / Production, purification and characterization of the antibacterial peptide produced by a strain of Bacillus sp. P34

Uma bactéria identificada como Bacillus sp. P34 isolada de intestino de peixe (Leporinus sp.) da Bacia Amazônica foi estudada quanto a sua capacidade de produzir substâncias do tipo-bacteriocina. As condições ótimas para produção da substância antimicrobiana foram determinadas. A produção da atividade antimicrobiana foi observada começando na fase exponencial de crescimento, sendo a atividade máxima observada no início da fase estacionária. Os resultados da Análise de Superfície de Resposta mostraram que a máxima produção da atividade antimicrobiana ocorreu a pH inicial entre 6.0 e 8.0 e temperaturas entre 25 e 37°C. A substância inibiu bactérias patogênicas e deteriorantes importantes em alimentos como Listeria monocytogenes, Bacillus cereus, Aeromonas hydrophila, Erwinia carotovora e Pasteurella haemolytica. O teste de termoestabilidade mostrou a perda de atividade quando a temperatura alcançou 100°C por 15 minutos. Foi sensível à ação das enzimas proteolíticas tripsina, papaína e pronase E. A substância antimicrobiana apresentou efeito bactericida e bacteriolítico sobre L. monocytogenes e B. cereus a 160 UA ml-1. O crescimento de Escherichia coli and Salmonella Enteritidis foi inibido somente quando o agente quelante EDTA foi adicionado juntamente. A atividade esporocida não foi observada. A análise da cultura de L. monocytogenes depois do tratamento com o composto antimicrobiano, usando espectroscopia de infravermelho com transformada de Fourier mostrou alterações no perfil de ácidos graxos e fosfolipídios da membrana celular bacteriana. Há evidências de que seu modo de ação interfira na membrana e na parede celular. A substância foi purificada pelo seguinte protocolo: precipitação com sulfato de amônio, cromatografias de gel filtração e de troca iônica. O peso molecular da substância foi determinado por espectroscopia de massas sendo 1498.68 Da. A substância antimicrobiana purificada apresentou sensibilidade ao tratamento com proteases e manutenção da atividade foi observada após congelamento e à incubação de 70°C por 30 minutos

A bacterium identified as Bacillus sp. strain P34 isolated from fish intestine (Leporinus sp.) from the Amazon basin was studied in its capacity to produce bacteriocinlike substances. The optimal conditions for producing the antimicrobial activity have been established. The antimicrobial activity was produced starting at the exponencial growth phase, and maximum activity was observed at early stationary phase. Response-surface data showed that maximum antimicrobial activity production was at initial pH between 6.0 and 8.0 and temperature between 25 and 37°C. The antimicrobial substance inhibited pathogenic and spoilage food bacteria such as Listeria monocytogenes, Bacillus cereus, Aeromonas hydrophila, Erwinia carotovora and Pasteurella haemolytica. The thermoestability test showed the loss of activity when the temperature reached 100°C for 15 min. It was sensitive to the proteolytic action of trypsin, papain and pronase E. The antimicrobial substance was bactericidal and bacteriolytic to L. monocytogenes and B. cereus at 160 AU ml-1. Growth of Escherichia coli and Salmonella Enteritidis was inhibited, but only when the chelating agent EDTA was co-added. Sporocidal activity was not observed. The analysis of the culture of L. monocytogenes after being treated with antimicrobial compound, using Fourier transform infrared spectroscopy, established a change in the profile that corresponding assignments of fatty acid and phospholipids. There was evidence that its mode of action to interfere with cell membrane and the cell wall. The substance was purified by the following protocol: precipitation with ammonium sulphate, gel filtration and ion exchange chromatography. The molecular weight was determined by mass spectroscopy as 1498.68 Da. Purified antimicrobial substance has shown sensitivity to protease treatment and maintained activity after freezing and incubation at 70°C for 30 min