In vitro algicidal effect of polypyrrole on Prototheca species isolates from bovine mastitisAlgicidal activity of polypyrrole on Prototheca spp.

Algae of the genus Prototheca are microorganisms involved in the occurrence of diseases in humans and animals. In bovine species, Prototheca spp. cause environmental mastitis, productive losses in dairy herds, mainly leading to the discard of infected cows. Currently, there are no effective anti-Prototheca spp. drugs to combat this infection. Thus, the search for an efficacious therapy for Prototheca spp. infections have become essential. Highly soluble polypyrrole (Ppy) is a molecule with known antimicrobial activity. This study aimed to characterize Prototheca spp. isolates from bovine mastitis as well as to evaluate the susceptibility profile and to verify the morphological alterations on Prototheca spp. isolates treated with Ppy. In this research, 36 Brazilian isolates of Prototheca spp. were characterized by restriction fragment length polymorphism polymerase chain reaction (RFLP-PCR) assay for the mitochondrial cytB gene. Additionally, Ppy algicidal activity against these isolates of Prototheca spp. was assessed by minimal microbicidal concentration method in microplates. Further, scanning electron microscopy (SEM) was performed in order to verify the morphological alterations on Prototheca spp. isolates in response to Ppy. The isolates were characterized as belonging to Prototheca zopfii genotype 2 (35/36) and Prototheca blaschkeae (1/36). Ppy had an algicidal effect on all isolates tested at concentrations ranging from 15.625 µg ml-1 to 62.5 µg ml-1. SEM showed changes on planktonic and sessile P. zopfii, including a decrease of the number of cells with the presence of an amorphous substance involving the cells. The algicidal activity of Ppy suggests the therapeutic potential of this molecule in the prevention and treatment of Prototheca spp. in bovine mastitis.

Antimicrobial activity of polypyrrole nanoparticles and aqueous extract of Moringa oleifera against Staphylococcus spp. carriers of multi-drug efflux system genes isolated from dairy farms.

Our objectives were to identify genes of the multi-drug efflux system and to evaluate the antimicrobial activities of polypyrrole nanoparticles (PPy-NPs) and aqueous extract of Moringa oleifera against Staphylococcus spp. isolated from dairy farms in Northeast Brazil. Initially, 162 Staphylococcus spp. isolates were subjected to in vitro antimicrobial sensitivity tests. Of these, 35 presented antimicrobial multi-drug resistance phenotypes. These 35 isolates were then referred for the detection of norA, norB, norC, msrA, mgrA, tet-38, and lmrS genes, all of which feature in multi-drug efflux systems. In the isolates carrying the genes, the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of PPy-NPs and Moringa oleifera aqueous extract were determined. In the molecular analysis of the 35 isolates norA, norC, tet-38, and msrA genes were detected and for the other genes norB, lmrS and mgrA there was no amplification. Antimicrobial activity was verified of PPy-NPs and aqueous extract of Moringa oleifera in Staphylococcus spp. carrying multi-drug efflux system genes. We concluded that there are multi-drug efflux system genes present in the Staphylococcus spp. from the agricultural environment in Northeast Brazil, and that aqueous extract of Moringa oleifera and PPy-NPs show bactericidal activity against these isolates.

Antimicrobial resistance profile of non-aureus Staphylococci isolates from buffalo, goat and sheep mastitis in the Northeast region of Brazil.

The study described in this Research Communication investigated the genotypic and phenotypic profiles of resistance to beta-lactams and other antimicrobials in non-aureus Staphylococci (NAS) isolated from buffalo, goat and sheep mastitis in the Northeast region of Brazil. A total of 190 isolates were analyzed and 42.3, 43.9 and 23.6% of them were positive for blaZ gene in buffalo, goat and sheep, respectively. Regarding the animal groups, in goats, amoxicillin was the antimicrobial with highest resistance index (72.7%), followed by penicillin G in buffaloes (51.9%) and ampicillin in sheep (43.1%). With regard to multiple antimicrobial resistance, 30.8% of NAS isolates from buffalo milk samples, 25.8% from goats and 25.0% from sheep presented multidrug-resistance. In the minimum inhibitory concentration (MIC) technique, amoxicillin MIC50 and MIC90 were 64 and 128 µg/ml, respectively, among isolates of the three animal species. In conclusion, high rates of resistance to beta-lactams are presented among NAS isolated from mastitis cases in buffaloes, goats and sheep in Northeast region of Brazil. These results provide an alert to animal and human health researchers, suggesting that the frequency of NAS needs to be reduced because they carry resistance genes which might increase the existing levels of antimicrobial resistance.

Multiplex polymerase chain reaction to identify and determine the toxigenicity of Corynebacterium spp with zoonotic potential and an overview of human and animal infections.

Corynebacterium diphtheriae, Corynebacterium ulcerans and Corynebacterium pseudotuberculosis constitute a group of potentially toxigenic microorganisms that are related to different infectious processes in animal and human hosts. Currently, there is a lack of information on the prevalence of disease caused by these pathogens, which is partially due to a reduction in the frequency of routine laboratory testing. In this study, a multiplex polymerase chain reaction (mPCR) assay that can simultaneously identify and determine the toxigenicity of these corynebacterial species with zoonotic potential was developed. This assay uses five primer pairs targeting the following genes: rpoB (Corynebacterium spp), 16S rRNA (C. ulcerans and C. pseudotuberculosis), pld (C. pseudotuberculosis), dtxR (C. diphtheriae) and tox [diphtheria toxin (DT) ]. In addition to describing this assay, we review the literature regarding the diseases caused by these pathogens. Of the 213 coryneform strains tested, the mPCR results for all toxigenic and non-toxigenic strains of C . diphtheriae, C. ulcerans and C. pseudotuberculosis were in 100% agreement with the results of standard biochemical tests and PCR-DT. As an alternative to conventional methods, due to its advantages of specificity and speed, the mPCR assay used in this study may successfully be applied for the diagnosis of human and/or animal diseases caused by potentially toxigenic corynebacterial species.

Genomic Characterization of Aeromonas veronii Provides Insights into Taxonomic Assignment and Reveals Widespread Virulence and Resistance Genes throughout the World.

Aeromonas veronii is a Gram-negative bacterial species that causes disease in fish and is nowadays increasingly recurrent in enteric infections of humans. This study was performed to characterize newly sequenced isolates by comparing them with complete genomes deposited at the NCBI (National Center for Biotechnology Information). Nine isolates from fish, environments, and humans from the São Francisco Valley (Petrolina, Pernambuco, Brazil) were sequenced and compared with complete genomes available in public databases to gain insight into taxonomic assignment and to better understand virulence and resistance profiles of this species within the One Health context. One local genome and four NCBI genomes were misidentified as A. veronii. A total of 239 virulence genes were identified in the local genomes, with most encoding adhesion, motility, and secretion systems. In total, 60 genes involved with resistance to 22 classes of antibiotics were identified in the genomes, including mcr-7 and cphA. The results suggest that the use of methods such as ANI is essential to avoid misclassification of the genomes. The virulence content of A. veronii from local isolates is similar to those complete genomes deposited at the NCBI. Genes encoding colistin resistance are widespread in the species, requiring greater attention for surveillance systems.

Genome Sequence of Pseudomonas sp. Strain LAP_36, A Rhizosphere Bacterium Isolated from King George Island, Antarctica.

Pseudomonas sp. strain LAP_36 was isolated from rhizosphere soil from Deschampsia antarctica on King George Island, South Shetland Islands, Antarctica. Here, we report on its draft genome sequence, which consists of 8,794,771 bp with 60.0% GC content and 8,011 protein-coding genes.

Efficacy assessment of an intramammary formulation based on soluble polypyrrole in cows with experimentally induced mastitis / Avaliação da eficácia de uma formulação intramamária à base de polipirrol solúvel em vacas com mastite experimentalmente induzida

Considerando a necessidade de terapias alternativas para a mastite bovina, uma inflamação, normalmente de causa infecciosa, de alta prevalência e de alto impacto econômico nas fazendas leiteiras, avaliou-se a eficácia antimicrobiana, in vivo, de uma formulação intramamária à base de polipirrol (PPy) solúvel, um polímero condutor promissor para aplicações biomédicas, especialmente como potente antibacteriano. Neste ensaio, quartos mamários de vacas holandesas sadias (n = 8) foram inoculados com Staphylococcus aureus e tratados, por via intramamária, com formulação experimental à base de PPy solúvel (5%) e com formulação comercial à base de sulfato de gentamicina. O efeito desses tratamentos foi avaliado com a realização de lactoculturas, da Contagem Bacteriana Total (TBC), da Contagem de Células Somáticas (SCC) e da análise da composição do leite das amostras obtidas de quartos mamários, em sete momentos experimentais. Avaliação hematológica dos animais também foi realizada. A aplicação intramamária de três doses da formulação experimental à base de PPy solúvel resultou em maiores log/mL da TBC e da SCC quando comparadas ao grupo controle positivo e ao grupo que recebeu sulfato de gentamicina. A administração da formulação experimental não induziu alterações na composição do leite e nos parâmetros hematológicos. Alguns fatores farmacocinéticos e farmacodinâmicos do PPy solúvel podem ser atribuídos a ineficácia antimicrobiana da pomada experimental. Outras pesquisas devem ser realizados em prol do desenvolvimento de formulações que permitam a atuação antibacteriana do PPy solúvel no ambiente intramamário de vacas leiteiras.

Given the need for alternative therapies for bovine mastitis, an infectious disease of high prevalence and significant economic impact in dairy farms, this study evaluated the in vivo antimicrobial efficacy of an intramammary formulation based on soluble polypyrrole (PPy), a promising conductive polymer for biomedical applications, especially as an effective antimicrobial agent. In this assay, mammary quarters of healthy Holstein Friesian cows (n = 8) were inoculated with Staphylococcus aureus and treated, via the intramammary route, with an experimental formulation based on soluble PPy (5%) and a commercial formulation based on gentamicin sulfate. The effect of these treatments was evaluated based on milk cultures, Total Bacterial Count (TBC), Somatic Cell Count (SCC), and milk composition analysis of mammary quarter samples in seven experimental moments. In addition, hematological evaluations of the animals were also performed. The intramammary application of three levels of the experimental formulation based on soluble PPy resulted in higher log/mL of the TBC and SCC compared to a positive control group and a group that received gentamicin sulfate. The experimental formulation did not induce changes in milk composition or in hematological parameters. Certain factors related to pharmacokinetics, such as the type of carrier used in the formulation and the pharmacodynamics of soluble PPy, may have contributed to the antimicrobial ineffectiveness of the experimental formulation. The results of this study neither define nor decrease the antimicrobial potential of soluble PPy. Further research is required to develop formulations that enable the antibacterial action of soluble PPy in the intramammary environment of dairy cows.

Biosecurity in dairy goat farms / Biosseguridade em propriedades de caprinos leiteiros

The goat milk industry has unquestionably grown in recent years due to the high demand for dairy products, which are considered nutritious and hypoallergenic. As a result, security measures are required in the production chain to provide consumers with safe products, although the concept of biosecurity is still incipient in Brazilian goat farming. Therefore, this study aimed to review the main biosecurity measures applied to dairy goat farms and suggest a program that contemplates these measures to promote animal health and welfare, given that biosecurity bolsters environmental sustainability and improves this agricultural sector. Biosecurity programs (BP) are composed of a set of measures and procedures aimed at herd health and applied in all stages of animal husbandry, interacting with different sectors that make up the production system and providing a set of policies and operational norms to protect herds against potentially pathogenic agents. Thus, BP require planning, execution, monitoring, audits, updates, understanding of the procedures, and awareness of those involved in the production chain. In addition, it includes continuing education programs and the development of contingency plans for specific emergencies. The information included in this study provides input to dairy goat farms to implement practices that improve the productivity of this agricultural sector.

A caprinocultura leiteira é um setor produtivo em crescimento, devido à alta demanda dos produtos lácteos considerados nutritivos e hipoalergênicos. Em consequência disso, são exigidas medidas de segurança na cadeia produtiva a fim de fornecer produtos inócuos ao consumidor. No entanto, os conceitos de biosseguridade ainda são incipientes na caprinocultura brasileira. Desta forma, este estudo tem a finalidade de revisar as principais medidas de biosseguridade aplicadas à caprinocultura leiteira e sugerir um programa que contemple estas medidas visando promover a saúde e o bem-estar animal. Além disso, a biosseguridade propicia a sustentabilidade do meio ambiente e potencializa melhorias neste setor agropecuário. Salienta-se que um programa de biosseguridade (PB) é composto por um conjunto de medidas e procedimentos de atenção à saúde do plantel, aplicados em todas as etapas da criação animal, interagindo com diversos setores que compõe o sistema produtivo, além de propiciar a implantação de um conjunto de políticas e normas operacionais, com o objetivo de proteger os rebanhos contra a introdução de qualquer agente infeccioso potencialmente patogênico. O PB necessita de planejamento, execução, monitoramento, auditorias, atualizações, bem como demanda de compreensão dos procedimentos e da sensibilização de todos os envolvidos nos processos da cadeia produtiva. Adicionalmente, inclui programas de educação continuada de todos os indivíduos e a elaboração de planos de contingência para situações emergenciais específicas. As informações incluídas neste estudo fornecerão aporte às propriedades de caprinocultura de leite para a implementação de ações que promovam melhorias na produtividade deste setor agropecuário.

Nanotechnology in veterinary medicine: a review / Nanotecnologia em medicina veterinária: uma revisão

The purpose of this review was to address the applicability of nanotechnology in veterinary medicine, with an emphasis on research in Brazil from 2013 to 2020. Firstly, we introduced to the general aspects of applicability of nanotechnology in veterinary medicine, and lately we pointed the research involving nanoscience performed in Brazil, in the studied period. Nanotechnology is the field of science that has the capacity to organize matter in nanoscale structures (1 to 100 nm), enabling innovations in different areas including biotechnology, agriculture, disease diagnosis, food and clothing industry, electronics, and pharmacological therapies. In veterinary medicine, several studies are being carried out in the world, mainly in the areas that involve search of new treatment options and the development of immunotherapy, as well as in the diagnosis of diseases. In Brazil, it is clear that the use of nanotechnology in veterinary medicine is still incipient, but it can be considered a growing area. In addition, several points have to be reflected and researched, including some adverse effects and implications to validate the safe use of nanotechnology in veterinary medicine. Therefore, this review highlighted the nanotechnology as a promise alternative in the current context of Brazilian technological innovation involving animal health, as well as a possible diagnostic tool and highlighting its potential therapeutic use in disease control in veterinary medicine. Regarding future perspectives, we believed that greater investment in science and technology could contribute to the advancement and strengthening of nanotechnology in Brazil.

O objetivo desta revisão foi abordar a aplicabilidade da nanotecnologia na medicina veterinária, com ênfase nas pesquisas no Brasil de 2013 a 2020. Primeiramente, apresentam-se os aspectos gerais da aplicabilidade da nanotecnologia em medicina veterinária e, posteriormente, apontam-se as pesquisas envolvendo nanociência realizadas no Brasil, no período estudado. A nanotecnologia é o campo da ciência capaz de organizar a matéria em estruturas de nanoescala (1 a 100 nm), permitindo inovações em diferentes áreas, incluindo biotecnologia, agricultura, diagnóstico de doenças, indústria de alimentos e vestuário, eletrônica e terapias farmacológicas. Na medicina veterinária, diversos estudos estão sendo realizados no mundo, principalmente nas áreas que envolvem a busca de novas opções de tratamento e o desenvolvimento de imunoterapia, bem como no diagnóstico de doenças. No Brasil, está claro que o uso da nanotecnologia na medicina veterinária ainda é incipiente, mas pode ser considerada uma área em crescimento. Além disso, vários pontos devem ser refletidos e pesquisados, incluindo alguns efeitos adversos e implicações para validar o uso seguro da nanotecnologia na medicina veterinária. Nesta revisão, destaca-se a nanotecnologia como uma alternativa promissora no atual contexto de inovação tecnológica brasileira, envolvendo saúde animal, o diagnóstico e o potencial uso terapêutico no controle de doenças em medicina veterinária. Em relação às perspectivas futuras, acredita-se que maiores investimentos em ciência e tecnologia são fatores fundamentais a contribuir para o avanço e fortalecimento da nanotecnologia no Brasil.

Economic impact and epidemiological aspects of subclinical mastitis and caprine arthritis-encephalitis in the semiarid northeastern region of Brazil / Impacto econômico e aspectos epidemiológicos da mastite e artrite encefalite caprina na região semiárida do Nordeste Brasileiro

Dairy activity in the semi-arid region has gained prominence due to its profit potential; however, there are still sanitary problems that result in significant economic impact. Thus, this study aimed to assess the economic impact and to conduct an epidemiological study of caprine arthritis-encephalitis (CAE) and subclinical mastitis in dairy goat herds of the semi-arid northeastern region of Brazil. The study proceeded with the collection of blood (n = 351) and milk samples (n = 321) from 15 rural properties with up to 50 animals, risk factor assessment, economic profile analysis, and georeferencing of the properties. The blood serum obtained was subjected to the AGID test. The milk samples were cultured on blood agar, and the reading was performed 24 hours after incubation. Overall, 3.4% of the animals were seropositive for CAE, and 15.8% were diagnosed with subclinical mastitis. 71.95% of the positive animals belonged to properties with a semi-intensive rearing system, weekly cleaning of the facilities, and no pre- or post-dipping. Subclinical mastitis caused by the main agent Staphylococcus spp. was associated with the breed type of the animals, highlighting two risk factors: purebred animals (OR = 12.52) and the milking performed close the soil (OR = 18.07). A reduction of up to 35.34% was observed in the income of dairy properties, constituting a considerable economic impact. Due to these sanitary problems, the economic impact demonstrates the need for a sanitary management program focused on these infirmities.

A atividade leiteira na região semiárida vem ganhando expressividade devido seu potencial lucrativo, no entanto, ainda existem problemas sanitários que ocasionam de grande impacto econômico. Assim, objetivou-se dimensionar o impacto econômico e realizar o estudo epidemiológico da artrite encefalite caprina (CAE) e mastite subclínica, nos rebanhos caprinos leiteiros do semiárido nordestino. A pesquisa seguiu-se com a coleta das amostras de sangue (n=351) e leite (n= 321), ava-liação dos fatores de risco, análise do perfil econômico das propriedades e georreferenciamento. O soro sanguíneo obtido foi submetido ao teste de IDGA. As amostras de leite foram semeadas em ágar sangue e a leitura realizada 24 horas após incuba-ção. No total, 3,4%, dos animais foram soropositivos para CAE e 15,8% diagnosticados com mastite subclínica. 71,95% dos animais positivos pertenciam a propriedades com sistema de criação semi-intensivo, limpeza das instalações semanalmente e sem realização de pré e pós dipping. A mastite subclínica causada pelo principal agente Staphylococcus spp. mostrou-se asso-ciada ao padrão racial dos animais, sendo observado dois fatores de risco: padrão racial puro (OR = 12,52) e ordenha no solo (OR = 18,07). A redução nas receitas das propriedades leiteiras foi de até 35,34%, portanto, um relevante impacto econômico. Pode-se constatar a circulação do vírus da CAE nas propriedades rurais visitadas, além da presença de patógenos causadores da mastite com importância na área da saúde pública. O impacto econômico gerado em função da presença desses proble-mas sanitários demonstra a necessidade da implantação de um programa de manejo sanitário com foco nessas enfermidades.

Economic impact and epidemiological aspects of subclinical mastitis and caprine arthritis-encephalitis in the semiarid northeastern region of Brazil / Impacto econômico e aspectos epidemiológicos da mastite e artrite encefalite caprina na região semiárida do Nordeste Brasileiro

Dairy activity in the semi-arid region has gained prominence due to its profit potential; however, there are still sanitary problems that result in significant economic impact. Thus, this study aimed to assess the economic impact and to conduct an epidemiological study of caprine arthritis-encephalitis (CAE) and subclinical mastitis in dairy goat herds of the semi-arid northeastern region of Brazil. The study proceeded with the collection of blood (n = 351) and milk samples (n = 321) from 15 rural properties with up to 50 animals, risk factor assessment, economic profile analysis, and georeferencing of the properties. The blood serum obtained was subjected to the AGID test. The milk samples were cultured on blood agar, and the reading was performed 24 hours after incubation. Overall, 3.4% of the animals were seropositive for CAE, and 15.8% were diagnosed with subclinical mastitis. 71.95% of the positive animals belonged to properties with a semi-intensive rearing system, weekly cleaning of the facilities, and no pre- or post-dipping. Subclinical mastitis caused by the main agent Staphylococcus spp. was associated with the breed type of the animals, highlighting two risk factors: purebred animals (OR = 12.52) and the milking performed close the soil (OR = 18.07). A reduction of up to 35.34% was observed in the income of dairy properties, constituting a considerable economic impact. Due to these sanitary problems, the economic impact demonstrates the need for a sanitary management program focused on these infirmities.(AU)

A atividade leiteira na região semiárida vem ganhando expressividade devido seu potencial lucrativo, no entanto, ainda existem problemas sanitários que ocasionam de grande impacto econômico. Assim, objetivou-se dimensionar o impacto econômico e realizar o estudo epidemiológico da artrite encefalite caprina (CAE) e mastite subclínica, nos rebanhos caprinos leiteiros do semiárido nordestino. A pesquisa seguiu-se com a coleta das amostras de sangue (n=351) e leite (n= 321), ava-liação dos fatores de risco, análise do perfil econômico das propriedades e georreferenciamento. O soro sanguíneo obtido foi submetido ao teste de IDGA. As amostras de leite foram semeadas em ágar sangue e a leitura realizada 24 horas após incuba-ção. No total, 3,4%, dos animais foram soropositivos para CAE e 15,8% diagnosticados com mastite subclínica. 71,95% dos animais positivos pertenciam a propriedades com sistema de criação semi-intensivo, limpeza das instalações semanalmente e sem realização de pré e pós dipping. A mastite subclínica causada pelo principal agente Staphylococcus spp. mostrou-se asso-ciada ao padrão racial dos animais, sendo observado dois fatores de risco: padrão racial puro (OR = 12,52) e ordenha no solo (OR = 18,07). A redução nas receitas das propriedades leiteiras foi de até 35,34%, portanto, um relevante impacto econômico. Pode-se constatar a circulação do vírus da CAE nas propriedades rurais visitadas, além da presença de patógenos causadores da mastite com importância na área da saúde pública. O impacto econômico gerado em função da presença desses proble-mas sanitários demonstra a necessidade da implantação de um programa de manejo sanitário com foco nessas enfermidades.(AU)

Efficacy of three methods used to control staphylococcal mastitis in dairy goats / Eficácia de três métodos empregados no controle da mastite estafilocócica em cabras leiteiras

Goat and sheep farming in Brazil relies on an expressive herd, especially in the Northeast region of the country, where small ruminants constitute a relevant economic activity, with milk production being an important source of income in some dry areas of the Northeastern states. However, there are still some sanitary problems in the management of dairy goats, among which mastitis is highlighted, especially that of the subclinical type. Therefore, the aim of this study was to evaluate the use of antibiotics, exclusively or in combination with vaccines, in the control of sub-clinical mastitis in small ruminants. The study was conducted on a rural property located in Santa Maria da Boa Vista-PE. Fifty-three female Saanen dairy goats were allocated into four experimental groups. An antibiotic therapy with a commercial antimicrobial based on cloxacillin was used in group 1. Group 2 received the commercial vaccine indicated for herds with an incidence of Staphylococcus strains and the same antibiotic described for Group 1. Group 3 was evaluated for the exclusive use of vaccination, and Group 4 was the control group. A microbiological examination of the milk was carried out to confirm cases of mastitis. The percentage of udders, infected by coagulase-negative Staphylococcus spp. (CNS) before and after the antibiotic + vaccine treatment was 42.8 and 7.1%, respectively. It was observed that the cloxacillinbased antibiotic could be used alone or in combination with mastitis vaccines to control staphylococcal mastitis in small ruminants. Vaccination alone was not effective in reducing the number of mastitis cases; therefore, a cross reaction against CNS was not observed. Thus, further studies to determine the potential of vaccines to control sub-clinical mastitis in small ruminants are necessary.

A caprinovinocultura brasileira dispõe de um expressivo rebanho, principalmente na região Nordeste do país, onde os pequenos ruminantes constituem uma relevante atividade econômica sendo a produção de leite uma importante fonte renda em algumas zonas secas dos estados do Nordeste. No entanto, ainda existem alguns problemas de ordem sanitária no manejo de cabras leiteiras, dentre esses, tem-se a mastite, principalmente aquela do tipo subclínica. Portanto, objetivou-se com este trabalho avaliar o uso isolado ou combinação de antibióticos e vacinas no controle da mastite em pequenos ruminantes. O estudo foi realizado em uma propriedade localizada no município de Santa Maria da Boa Vista - PE. Um total de 53 fêmeas leiteiras Saanen foram divididas em quatros grupos experimentais. No grupo 01 foi utilizada a antibioticoterapia, com antimicrobiano comercial a base de cloxacilina. O grupo 02 recebeu vacina comercial indicada para rebanhos com incidência de cepas do gênero Staphylococcus e o mesmo antibiótico descrito para o grupo 01. Em um terceiro grupo apenas a vacina foi empregada, e por último, foi constituído um grupo controle. Empregou-se o exame microbiológico do leite para confirmação dos casos de mastite. A porcentagem de tetos infectados por Staphylococcus coagulase negativa (SCN) antes e após o tratamento Antibiótico + Vacina foi de 42,8 e 7,1%, respectivamente. Observou-se que o antibiótico a base de cloxacilina pode ser usado isoladamente ou em associação a vacinas anti-mastíticas para controle da mastite estafilocócica em pequenos ruminantes. O tratamento que preconizava a aplicação apenas da vacina não foi eficiente na redução dos casos da doença, portanto, não foi observada uma reação cruzada contra os SCN. Assim, faz-se necessária a continuidade dos estudos acerca do potencial das vacinas para o controle da mastite em pequenos ruminantes.

Phenotypic and genotypic characteristics and resistance profile of Staphylococcus spp. from bovine mastitis

Background: Bovine mastitis remains one of the health problems that cause the most damage to milk producers. The negative impact of mastitis is due to reduced milk production, early slaughter of females, reduced commercial value of the affected animals, losses in the genetic potential of the herd, expenses with medicines and veterinary medical assistance. Staphylococcus spp. stands out as the cause of this disease and has been able to remain in the mammary gland, becoming resistant to several antimicrobials. The aims of the present study were to characterize the phenotypes, genotypes and resistance profiles of Staphylococcus spp. isolates from bovine mastitis cases in the state of Pernambuco, Brazil. Materials, Methods & Results: These isolates were classified according to biochemical tests and the presence of the nuc gene. The polymerase chain reaction (PCR) for amplification of the mecA and blaZ genes was used to analyze the genetic potentials of antimicrobial resistance. Isolates were also phenotypically tested for resistance to nine antimicrobials (ampicillin, doxicillin, erythromycin, gentamicin, rifampicin, cephalothin, amoxicillin, nalidixic acid and oxacillin). The genetic potentials for biofilm production were evaluated by the amplifications of the icaD, icaA and bap genes. The phenotypic test of gentian violet was used for biofilm formation analyzes. Ninety-three (93.0%) of the isolates among the Staphylococcus spp. samples were classified as Staphylococcus aureus. The lowest percentage of sensitivity observed was for amoxicillin (28.0%). All of the isolates were sensitive to erythromycin and gentamicin, and 15 (15%) exhibited sensitivity to all of the drugs tested. All of the isolates were negative for the mecA gene, and 36 (36%) were positive for blaZ. In the adhesion microplate tests, 44 (44%) of the isolates were capable of biofilm formation...

Efficacy of three methods used to control staphylococcal mastitis in dairy goats / Eficácia de três métodos empregados no controle da mastite estafilocócica em cabras leiteiras

Goat and sheep farming in Brazil relies on an expressive herd, especially in the Northeast region of the country, where small ruminants constitute a relevant economic activity, with milk production being an important source of income in some dry areas of the Northeastern states. However, there are still some sanitary problems in the management of dairy goats, among which mastitis is highlighted, especially that of the subclinical type. Therefore, the aim of this study was to evaluate the use of antibiotics, exclusively or in combination with vaccines, in the control of sub-clinical mastitis in small ruminants. The study was conducted on a rural property located in Santa Maria da Boa Vista-PE. Fifty-three female Saanen dairy goats were allocated into four experimental groups. An antibiotic therapy with a commercial antimicrobial based on cloxacillin was used in group 1. Group 2 received the commercial vaccine indicated for herds with an incidence of Staphylococcus strains and the same antibiotic described for Group 1. Group 3 was evaluated for the exclusive use of vaccination, and Group 4 was the control group. A microbiological examination of the milk was carried out to confirm cases of mastitis. The percentage of udders, infected by coagulase-negative Staphylococcus spp. (CNS) before and after the antibiotic + vaccine treatment was 42.8 and 7.1%, respectively. It was observed that the cloxacillinbased antibiotic could be used alone or in combination with mastitis vaccines to control staphylococcal mastitis in small ruminants. Vaccination alone was not effective in reducing the number of mastitis cases; therefore, a cross reaction against CNS was not observed. Thus, further studies to determine the potential of vaccines to control sub-clinical mastitis in small ruminants are necessary.(AU)

A caprinovinocultura brasileira dispõe de um expressivo rebanho, principalmente na região Nordeste do país, onde os pequenos ruminantes constituem uma relevante atividade econômica sendo a produção de leite uma importante fonte renda em algumas zonas secas dos estados do Nordeste. No entanto, ainda existem alguns problemas de ordem sanitária no manejo de cabras leiteiras, dentre esses, tem-se a mastite, principalmente aquela do tipo subclínica. Portanto, objetivou-se com este trabalho avaliar o uso isolado ou combinação de antibióticos e vacinas no controle da mastite em pequenos ruminantes. O estudo foi realizado em uma propriedade localizada no município de Santa Maria da Boa Vista - PE. Um total de 53 fêmeas leiteiras Saanen foram divididas em quatros grupos experimentais. No grupo 01 foi utilizada a antibioticoterapia, com antimicrobiano comercial a base de cloxacilina. O grupo 02 recebeu vacina comercial indicada para rebanhos com incidência de cepas do gênero Staphylococcus e o mesmo antibiótico descrito para o grupo 01. Em um terceiro grupo apenas a vacina foi empregada, e por último, foi constituído um grupo controle. Empregou-se o exame microbiológico do leite para confirmação dos casos de mastite. A porcentagem de tetos infectados por Staphylococcus coagulase negativa (SCN) antes e após o tratamento Antibiótico + Vacina foi de 42,8 e 7,1%, respectivamente. Observou-se que o antibiótico a base de cloxacilina pode ser usado isoladamente ou em associação a vacinas anti-mastíticas para controle da mastite estafilocócica em pequenos ruminantes. O tratamento que preconizava a aplicação apenas da vacina não foi eficiente na redução dos casos da doença, portanto, não foi observada uma reação cruzada contra os SCN. Assim, faz-se necessária a continuidade dos estudos acerca do potencial das vacinas para o controle da mastite em pequenos ruminantes.(AU)

Antimicrobial potential of Jurema preta and umburana, native species of the Caatinga biome, on Staphylococcus isolated from small ruminants with mastitis / Potencial antimicrobiano de jurema preta e umburana, espécies nativas do bioma caatinga, frente a isolados de Staphylococcus provenientes de casos de mastite em pequenos ruminantes

Mastitis is responsible for both damage to animal health and economic losses to the industry. To identify effective treatments for this disease, products extracted from a variety of plants with antimicrobial potential have gained attention. The present study aimed to assess the in vitro antibacterial potential ofthe ethanolic extract of two plant species from the Caatinga biome against bacteria isolated from small ruminants with subclinical mastitis. The leaves of Mimosa tenuiflora and Commiphora leptophloeos were dried and processed to obtain crude ethanolic extracts and their phenolic composition was evaluated. In total, 33 Staphylococcus spp. isolates from the bacterial collection of the Laboratory of Food Quality Control of IF SERTÃO-PE were used for evaluation of biofilm production. Furthermore, an antimicrobial susceptibility test was conducted using the minimal bactericidal concentration (MBC) method against the two ethanolic extracts. The toxic potential was measured through a toxicity test with Artemia salina. The quantification of the phenolic compounds revealed that the ethanolic extracts of M. tenuiflora and C. leptophloeos possessed higher amounts of myricetin (43.2 and 294.9 mg in 10 g, respectively) in relation to the other compounds. A 39.4% positivity rate was observed in the nuc gene investigation. The biofilm production analysis revealed that 96.9% of the isolates produced biofilm, evidencing the evolution the microorganisms regarding the development of resistance mechanisms. The MBC results showed an inhibition range between 195.30 and 3125.00 µg mL-¹ and between 781.20 and 6250.00 µg mL-¹ for the extracts of M. tenuiflora and C. leptophloeos, respectively. The M. tenuiflora extract showed the highest activity, suppressing 100% of the bacterial isolates (n=26), whereas the extract of C. leptophloeos showed an inhibition percentage of 69.23%. The crude ethanolic extract (EEB) [...].(AU)

A mastite é responsável por danos à saúde animal e prejuízos financeiros ao produtor. Na busca por alternativas eficazes no controle da mastite, destacam-se os produtos naturais extraídos de diversas plantas que possuem ação antibacteriana. Neste contexto, o presente trabalho teve por objetivo avaliar o potencial antimicrobiano in vitro do extrato etanólico de duas espécies do bioma caatinga frente a bactérias isoladas de casos de mastite subclínica em pequenos ruminantes. As folhas de jurema preta e umburana de cambão foram secas e processadas para obtenção dos extratos etanólicos bruto para o estudo da composição fenólica. Foram utilizados 33 isolados de Staphylococcus spp. provenientes da bacterioteca do Laboratório de Controle de Qualidade de Alimentos do IF SERTÃO-PE. Com os isolados de Staphylococcus spp. realizou-se a análise de quantificação da produção de biofilme. Além disso, empregou-se o teste de sensibilidade antimicrobiana, utilizando-se a técnica da concentração bactericida mínima (CBM) frente aos dois extratos etanólicos. Para conhecer o potencial tóxico, utilizou-se o teste de toxicidade com Artemia salina. A análise de quantificação dos compostos fenólicos demonstrou que os extratos etanólicos de jurema preta e umburana de cambão apresentaram a miricetina em quantidades bastante elevadas, 43,2 e 294,9 mg em 10 g respectivamente, em relação aos demais compostos detectados. Na pesquisa do gene nuc, observou-se uma positividade de 39,4%.A análise da produção de biofilme revelou que 96,9% dos isolados apresentaram a capacidade de produção do biofilme, evidenciando a evolução destes micro-organismos quanto ao desenvolvimento de mecanismos de resistência. Os resultados referentes à concentração bactericida mínima indicaram uma faixa de inibição entre 195,30 [...].(AU)

Antimicrobial potential of Jurema preta and umburana, native species of the Caatinga biome, on Staphylococcus isolated from small ruminants with mastitis / Potencial antimicrobiano de jurema preta e umburana, espécies nativas do bioma caatinga, frente a isolados de Staphylococcus provenientes de casos de mastite em pequenos ruminantes

Mastitis is responsible for both damage to animal health and economic losses to the industry. To identify effective treatments for this disease, products extracted from a variety of plants with antimicrobial potential have gained attention. The present study aimed to assess the in vitro antibacterial potential ofthe ethanolic extract of two plant species from the Caatinga biome against bacteria isolated from small ruminants with subclinical mastitis. The leaves of Mimosa tenuiflora and Commiphora leptophloeos were dried and processed to obtain crude ethanolic extracts and their phenolic composition was evaluated. In total, 33 Staphylococcus spp. isolates from the bacterial collection of the Laboratory of Food Quality Control of IF SERTÃO-PE were used for evaluation of biofilm production. Furthermore, an antimicrobial susceptibility test was conducted using the minimal bactericidal concentration (MBC) method against the two ethanolic extracts. The toxic potential was measured through a toxicity test with Artemia salina. The quantification of the phenolic compounds revealed that the ethanolic extracts of M. tenuiflora and C. leptophloeos possessed higher amounts of myricetin (43.2 and 294.9 mg in 10 g, respectively) in relation to the other compounds. A 39.4% positivity rate was observed in the nuc gene investigation. The biofilm production analysis revealed that 96.9% of the isolates produced biofilm, evidencing the evolution the microorganisms regarding the development of resistance mechanisms. The MBC results showed an inhibition range between 195.30 and 3125.00 µg mL-¹ and between 781.20 and 6250.00 µg mL-¹ for the extracts of M. tenuiflora and C. leptophloeos, respectively. The M. tenuiflora extract showed the highest activity, suppressing 100% of the bacterial isolates (n=26), whereas the extract of C. leptophloeos showed an inhibition percentage of 69.23%. The crude ethanolic extract (EEB) [...].

A mastite é responsável por danos à saúde animal e prejuízos financeiros ao produtor. Na busca por alternativas eficazes no controle da mastite, destacam-se os produtos naturais extraídos de diversas plantas que possuem ação antibacteriana. Neste contexto, o presente trabalho teve por objetivo avaliar o potencial antimicrobiano in vitro do extrato etanólico de duas espécies do bioma caatinga frente a bactérias isoladas de casos de mastite subclínica em pequenos ruminantes. As folhas de jurema preta e umburana de cambão foram secas e processadas para obtenção dos extratos etanólicos bruto para o estudo da composição fenólica. Foram utilizados 33 isolados de Staphylococcus spp. provenientes da bacterioteca do Laboratório de Controle de Qualidade de Alimentos do IF SERTÃO-PE. Com os isolados de Staphylococcus spp. realizou-se a análise de quantificação da produção de biofilme. Além disso, empregou-se o teste de sensibilidade antimicrobiana, utilizando-se a técnica da concentração bactericida mínima (CBM) frente aos dois extratos etanólicos. Para conhecer o potencial tóxico, utilizou-se o teste de toxicidade com Artemia salina. A análise de quantificação dos compostos fenólicos demonstrou que os extratos etanólicos de jurema preta e umburana de cambão apresentaram a miricetina em quantidades bastante elevadas, 43,2 e 294,9 mg em 10 g respectivamente, em relação aos demais compostos detectados. Na pesquisa do gene nuc, observou-se uma positividade de 39,4%.A análise da produção de biofilme revelou que 96,9% dos isolados apresentaram a capacidade de produção do biofilme, evidenciando a evolução destes micro-organismos quanto ao desenvolvimento de mecanismos de resistência. Os resultados referentes à concentração bactericida mínima indicaram uma faixa de inibição entre 195,30 [...].

Bovine mastitis caused by multidrug-resistant Nocardia farcinica

Background: Mastitis caused by Nocardia is characterized by pyogranulomatous inflammation related to inadequatehygiene conditions and is difficult to treat. Prompted by the absence of documentation of Nocardia farcinica associated tobovine mastitis in the Northeast region of Brazil, this is the first report to describe bovine mastitis caused by multidrugresistant N. farcinica.Case: Four milk samples (one from each teat) obtained from a 3-year-old Jersey cow raised on a property located in the metropolitan region of Recife, Pernambuco state, Brazil, were submitted to the Laboratory of Infectious-Contagious Diseases ofthe Veterinary Hospital at Universidade Federal Rural de Pernambuco [UFRPE]. At the laboratory, samples were cultured inbase agar enriched with 7% sheep blood (blood agar) in a microbiological incubator at 37°C under aerobic conditions for 72h. After only 48 h, however, pure bacterial colony growth was observed in all samples. Macroscopic analysis revealed smallcolonies, with an irregular shape, dry aspect, and greyish in color. Gram-positive rods forming filaments and/or ramificationswere observed using a Gram staining method. Nocardia spp. were identified according to morphotinctorial characteristics.Susceptibility testing using the disc-diffusion method in agar (antibiogram) was performed using the following antibiotics:penicillin (10 IU), tetracycline (30 µg), amoxicillin (10 µg), gentamicin (10 µg), cephalexin (30 µg), erythromycin (15 µg),cephalothin (30 µg) and ampicillin (30 µg). However, the organism exhibited resistance to all drugs; as such, a new milksample was obtained at the same location the initial samples were collected. Samples (approximately 5 mL) were collectedaseptically and separately from all four teats in sterile bottles, during which the presence of granular material was noted.Bacterial culture was performed as...

Bovine mastitis caused by multidrug-resistant Nocardia farcinica

Background: Mastitis caused by Nocardia is characterized by pyogranulomatous inflammation related to inadequatehygiene conditions and is difficult to treat. Prompted by the absence of documentation of Nocardia farcinica associated tobovine mastitis in the Northeast region of Brazil, this is the first report to describe bovine mastitis caused by multidrugresistant N. farcinica.Case: Four milk samples (one from each teat) obtained from a 3-year-old Jersey cow raised on a property located in the metropolitan region of Recife, Pernambuco state, Brazil, were submitted to the Laboratory of Infectious-Contagious Diseases ofthe Veterinary Hospital at Universidade Federal Rural de Pernambuco [UFRPE]. At the laboratory, samples were cultured inbase agar enriched with 7% sheep blood (blood agar) in a microbiological incubator at 37°C under aerobic conditions for 72h. After only 48 h, however, pure bacterial colony growth was observed in all samples. Macroscopic analysis revealed smallcolonies, with an irregular shape, dry aspect, and greyish in color. Gram-positive rods forming filaments and/or ramificationswere observed using a Gram staining method. Nocardia spp. were identified according to morphotinctorial characteristics.Susceptibility testing using the disc-diffusion method in agar (antibiogram) was performed using the following antibiotics:penicillin (10 IU), tetracycline (30 µg), amoxicillin (10 µg), gentamicin (10 µg), cephalexin (30 µg), erythromycin (15 µg),cephalothin (30 µg) and ampicillin (30 µg). However, the organism exhibited resistance to all drugs; as such, a new milksample was obtained at the same location the initial samples were collected. Samples (approximately 5 mL) were collectedaseptically and separately from all four teats in sterile bottles, during which the presence of granular material was noted.Bacterial culture was performed as...(AU)

Biofilm formation by Prototheca zopfii isolated from clinical and subclinical bovine mastitis in distinct growth conditions under different dyes / Formação de biofilme por Prototheca zopfii isolados de mastite bovina clínica e subclínica em distintas condições de crescimento sob diferentes corantes

ABSTRACT: Prototheca spp. have been reported as an emergent environmental mastitis pathogen in several countries. Biofilm formation is a significant factor associated with different degrees of virulence developed by many microorganisms, including Prototheca spp. The present study aimed to compare two growth conditions and two staining dyes to determine which combination was more appropriate to evaluate qualitatively and quantitatively the production of biofilm by P. zopfii. Biofilm formation was evaluated in polystyrene microplates under static and dynamic growth conditions and staining with crystal violet or cotton blue dye. All P. zopfii isolates from cows with mastitis were classified as biofilm-producers in all growth conditions and staining. The cotton blue dye proved to be more appropriate method to classify the intensity of P. zopfii biofilm production.

RESUMO: Prototheca spp. tem sido relatado como um patógeno ambiental causador de mastite bovina em vários países. A formação de biofilme é um fator associado a diferentes graus de virulência desenvolvidos por muitos microrganismos, incluindo Prototheca zopfii. O presente estudo teve como objetivo comparar duas condições de crescimento e dois corantes para determinar a combinação mais adequada para avaliar qualitativa e quantitativamente a produção de biofilme por P. zopfii. A formação de biofilme foi avaliada em microplacas de poliestireno sob condições estáticas e dinâmicas de crescimento e coloração com cristal violeta ou azul de algodão. Todos os isolados de P. zopfii de vacas com mastite foram caracterizados como produtores de biofilme, independentemente das condições de crescimento e coloração. O corante azul de algodão demonstrou ser o método mais adequado para classificar a intensidade de produção de biofilme de P. zopfii.

Minimum bactericidal concentration of commercial disinfectants on Staphylococcus spp. isolated from mastitis in goats and detection of the icaD gene / Concentração bactericida mínima de desinfetantes comerciais frente à Staphylococcus spp. isolados de mastite em caprinos e detecção do gene icaD

This study aimed to evaluate the in vitro efficacy of commercial disinfectants on Staphylococcus spp., isolated from mastitis cases in goats, and to associate the observed resistance with the presence of the icaD gene. Broth microdilution was employed to evaluate the in vitro antimicrobial activity of the disinfectants, whereas the Congo Red technique was used for the evaluation of biofilm production and amplification of the icaD gene. All evaluated samples were sensitive to disinfectants, with the following ranges of activity: quaternary ammonium (0.13 - 21.33 µg/ml), chlorhexidine (4.00 - 313.00 µg/ml) and iodine (190.00 - 12500.00 µg/ml); however, the sodium hypochlorite-based disinfectant showed no bactericidal activity in the concentration range from 15.0 to 0.03 µg/ml. The icaD gene presented a frequency of 14.7% in the isolate samples. Fisher’s exact test showed a significant effect of the relation between the minimum bactericidal concentration value of the quaternary ammonium-based disinfectant and the presence/absence of the icaD gene (P <0.01). Commercial disinfectants with quaternary ammonium, chlorhexidine and iodine active ingredients presented in vitro activity even at concentrations lower than those recommended by the manufacturers. Therefore, the periodic evaluation of the sensitivity profile of the disinfectants must be performed.

O presente trabalho objetivou avaliar a eficácia in vitro de desinfetantes comerciais frente à Staphylococcus spp. isolados de casos de mastite em cabras e, relacionar a resistência observada com a presença do gene icaD. A microdiluição em caldo foi utilizada para avaliar a atividade antimicrobiana in vitro dos desinfetantes, enquanto a técnica de vermelho congo para a avaliação da produção de biofilme e amplificação do gene icaD. Todas as amostras avaliadas foram sensíveis aos desinfetantes, com as seguintes faixas de atividade: amônia quaternária (0,13 - 21,33 µg/ml), clorexidina (4,00 - 313,00 µg/ml) e iodo (190,00 - 12500,00 µg/ml), no entanto o desinfetante a base de hipoclorito de sódio não apresentou atividade bactericida na faixa de concentração 15,0 a 0,03 µg/ml. O gene icaD apresentou frequência de 14,7% nos isolados. O teste exato de Fisher apontou efeito significativo da relação entre o valor da concentração bactericida mínima do desinfetante à base de amônia quaternária e a presença/ausência do gene icaD (P<0,01). Os desinfetantes comerciais com os princípios ativos amônia quaternária, clorexidina e iodo, apresentaram atividade in vitro até mesmo em concentrações inferiores às preconizadas pelos fabricantes o que torna necessária a realização periódica da avaliação do perfil de sensibilidade dos desinfetantes.