Antimicrobial efficiency of bromhexine hydrochloride against endometritis-causing Escherichia coli and Trueperella pyogenes in bovines.

In this study, the main agents associated with endometritis in cows in the state of Santa Catarina, Brazil, were identified and the resistance profile and virulence mechanisms of the bacterial isolates were evaluated. Isolates of Escherichia coli and Trueperella pyogenes were tested for their biofilm forming ability and the antimicrobial action of bromhexine hydrochloride in combination with other antimicrobials. A total of 37 uterine lavage samples were collected from cows with endometritis. Of the 55 bacteria isolated, 25.4% were identified as T. pyogenes and 16.3% as E. coli. The bacterial isolates showed greater resistance to sulfamethoxazole + trimethoprim (58.2%) and tetracycline (56.3%). Among the species, E. coli showed the highest resistance rates, with 100% of isolates showing resistance to amoxicillin, streptomycin, and gentamicin. The results of the minimum inhibitory concentration for the T. pyogenes isolates showed that 91.6% of the isolates were resistant to enrofloxacin and tetracycline, and 75% were resistant to ceftiofur and sulfamethoxazole + trimethoprim. All E. coli and T. pyogenes isolates showed biofilm forming ability. The plo, fimA, and nanH genes were identified in 100% of T. pyogenes isolates. In parallel, 100% of E. coli isolates had the fimH gene, and 11.1% had the csgD gene. Bromhexine hydrochloride showed antimicrobial activity against 100% of E. coli isolates and 66.6% of T. pyogenes isolates. Furthermore, when associated with antimicrobials, bromhexine hydrochloride has a synergistic and additive effect, proving to be an option in the treatment of endometritis in cows and an alternative for reducing the use of antimicrobials.

Synergy between polypyrrol and benzoic acid against antibiotic-resistant Salmonella spp.

AIMS: The purpose was to characterize Salmonella Heidelberg (SH) and Minnesota (SM) isolates in terms of their resistance and persistence profile and to assess the antimicrobial effect of benzoic acid (BA) and polypyrrole (PPy). METHODS AND RESULTS: The 20 isolates from broiler litter drag swabs were submitted to antibiogram and efflux pump expression. The minimum inhibitory/bactericidal concentration (MIC/MBC) of the compounds, synergistic activity, time kill, biofilm production, presence of related genes, and molecular docking between compounds and bacterial target sites were evaluated. All isolates showed multidrug resistance (MDR) and BA and PPy showed mean MIC (1750 and 342 µg ml-1) and MBC (3167 and 1000 µg ml-1), respectively. None of the isolates expressed an efflux pump. The compounds showed synergism against an SH isolate and reduced the count by 3 logs in the presence of the compounds after 4 h. Most isolates (16/20) produced weak to moderate biofilm and 17 showed genes related to biofilm. The compounds interacted with two essential proteins, 3,4-dihydroxy-2-butanone 4-phosphate synthase proteins and ferritin-like domain-containing protein, in bacterial metabolism at different target sites. CONCLUSIONS: It can be concluded that BA and PPy showed activity on SH and SM, MDR, and biofilm producers, with a potential synergistic effect.

Effect of a phytogenic feed additives mixture on milk physico-chemical properties and biochemical parameters of Holstein cows / Efeito de uma mistura de aditivos fitogênicos alimentares nas propriedades físico-químicas do leite e parâmetros bioquímicos de vacas leiteiras da raça holandês

ABSTRACT: This study aimed to analyze the effect of phytogenic additives based on cardol, cardanol, and ricinoleic acid on the physico-chemical qualities and biochemical parameters of Holstein cow milk. Nineteen animals were divided into the control (GI) and treatment (GII) groups. Prior to the beginning of the experiment, sodium monensin was removed from the feed provided to the animals. This study consisted of two phases. In the first, animals from GI received the standard diet, without additives, while animals from GII received the standard diet supplemented with 10 g of phytogenic additives for 60 days. At the end of the first stage, the animals remained 30 days without receiving additives and in the second phase the groups were inverted, and the GII again received the phytogenic additive during the subsequent 60 days. Physical-chemical analyzes of milk were performed at moments 0, 15, 30, 45 and 60 days of the experiment and the blood parameters at times 0 and 60 days of the experiment were evaluated. There was no significant difference for the treatment x time interaction in any of the variables, but there was a trend between groups for the CCS parameter and there was a statistical difference for the protein, as well as for the AST. In this study it is observed that the addition of phytogenic additive modulating rumen fermentation based on cardol, cardanol and ricinoleic acid does not compromise, in general, the quality of milk, with positive results for some specific parameters such as protein content and aminotransferase.

RESUMO: Esse estudo teve por objetivo avaliar o efeito de um aditivo fitogênico a base de cardol, cardanol e ácido ricinoléico na qualidade físico-química do leite e perfil bioquímico de vacas da raça Holandês. Dezenove animais foram divididos em dois grupos: Grupo Controle (GI) e Grupo Tratado (GII). Previamente ao início do experimento, retirou-se a monensina sódica da ração fornecida aos animais. Este estudo consistiu de duas fases, sendo que na primeira fase, os animais do GI receberam a dieta padrão, sem aditivos, enquanto os animais do grupo GII receberam a ração padrão suplementada com 10 g dos aditivos fitogênicos durante 60 dias. Ao final da primeira etapa, os animais permaneceram 30 dias sem receber aditivos e na segunda fase os grupos foram invertidos, sendo que o GII novamente recebeu o aditivo fitogênico durante os 60 dias subsequentes. Foram realizadas análises físico-químicas do leite nos momentos 0, 15, 30, 45 e 60 dias do experimento e avaliados os parâmetros sanguíneos nos momentos 0 e 60 dias do experimento. Não se observou diferença significativa para a interação tratamento x tempo em nenhuma das variáveis, mas tendência entre grupos para o parâmetro de CCS, e houve diferença estatística para a proteína, assim como para a AST. Neste estudo observa-se que a adição de aditivo fitogênico modulador de fermentação ruminal a base de cardol, cardanol e ácido ricinoléico não compromete, de forma geral, a qualidade do leite, tendo resultados positivos para alguns parâmetros específicos como teor de proteína e aspartato aminotransferase.

Subclinical mastitis in Lacaune sheep: Causative agents, impacts on milk production, milk quality , oxidative profiles and treatment efficacy of ceftiofur.

Mastitis is a major disease affecting dairy sheep. It is caused by microorganisms that generate inflammation of the mammary gland in response to tissue invasion. This syndrome affects the welfare of ewes, as well as the production and quality of the milk, thereby reducing its productive efficiency. Because mastitis causes inflammation process, it also increases the production of free radicals that cause lesions via lipoperoxidation, causing damage to proteins, cells and tissues. One way to minimize the impact of the disease is antimicrobial treatment. Nevertheless, the continuous use of antimicrobials contributes to microbial resistance, in addition to producing residues in the milk and derivatives if not given during the grace period. Therefore, the objective of this study was to evaluate the consequences of subclinical mastitis on ewe health, milk production, milk composition and quality. We also evaluated the susceptibility of the bacteria in vitro using disk diffusion antibiograms. Finally, we performed two-way testing of efficacy of treatment in Lacaune ewes using the same agents. In the first stage of the study, 30 lactating ewes (±90 days) were used, 10 of which were negative on the CMT (California Mastitis Test) used as control group (CG) and 20 sheep with subclinical mastitis diagnosed by CMT (MG). Samples were collected and several analyses were performed on the milk and blood. We found that ewes in the MG had higher lipid peroxidation in serum and milk, as well as lower production, with reduction of the total dry extract in milk. There were 15 isolates of Staphylococcus hyicus, four isolates of each S. epidermidis and S. intermedius, and two isolates of Corynebacterium spp. The primary hematological result was leukocytosis in ewes with mastitis. Based on the antibiogram, we chose ceftiofur for in vivo tests. In this stage, we divided the sheep with subclinical mastitis into two subgroups of 10 ewes each, to receive drug by two routes: intramuscular (IM) and intramammary (IMM). In the IMM group, of the 10 CMT-positive ewes at the beginning of the experiment, seven were already negative by the racket test 120 h after the last application (70% efficacy). In the IM group, of the 10 positive ewes, only four were negative after 120 h of the final application, a low efficacy treatment (40%). We evaluated antimicrobial residues in the milk of treated animals. We found this material within 5 days after treatment in the two forms used; despite the fact that the product's stated withholding period is 3 days. We conclude that ewes with mastitis produce less milk of lower quality. We also conclude that, although ceftiofur is 100% effective in vitro, when used in ewes with mastitis, the efficacy did not exceed 70%, and was more efficient when administered via the intramammary route.

High prevalence of porcine circovirus 2, porcine parvovirus, and pathogenic leptospires in mummified swine fetuses in Southern Brazil / Alta prevalência de parvovírus suíno, circovírus suíno 2 e leptospiras patogênicas em fetos suínos mumificados na região Sul do Brasil

ABSTRACT: Modern swine production employs a high degree of technology and is organized in various production stages, in which reproduction is one of the most significant. However, reproductive losses associated to fetal death are still high. Fetal losses, including fetal mummification, may occur at a rate below 1.5% in a sound herd. The causes of fetal death can be very diverse, and include various infectious agents. Therefore, investigating the causes of fetal losses is important in the decision-making process related to the control and prophylaxis of the herd. The goal of this study was to carry out a molecular survey, analyzing porcine circovirus 2 (PCV2), porcine parvovirus (PPV), and pathogenic Leptospira as potential agents related to fetal death in swine farms in Southern Brazil. We collected samples from three farms with a mummified index ≥ 2.5%. Fragments of brain, lung, kidney, liver, and heart were sampled for PCR analysis. Out of 100 samples examined, 87, 68, and 22 were positive for PCV2, PPV, and Leptospira spp., respectively. Moreover, we also identified coinfections with two and three pathogens in the same sample. Our findings contribute to the spreading of scientific knowledge related to infectious causes (PPV, PCV2, and pathogenic leptospires) of fetal losses in swine, as evidenced by the high frequencies of the investigated agents.

RESUMO: A produção moderna de suínos emprega um alto grau de tecnologia e é organizada em várias etapas de produção, nas quais a reprodução é uma das mais importantes. No entanto, as perdas reprodutivas associadas à morte fetal ainda são altas. Perdas fetais, incluindo mumificação, podem ocorrer em taxa abaixo de 1,5% em rebanhos sadios. As causas da morte fetal podem ser diversas, incluindo causas infecciosas. Portanto, a investigação de causas relacionadas à ocorrência de perdas fetais é importante no processo relacionado ao controle e profilaxia do rebanho. O objetivo deste estudo foi realizar um levantamento molecular, incluindo circovírus suíno 2 (PCV2), parvovírus suíno (PPV), e leptospiras patogênicas como potenciais agentes relacionados à morte fetal em suínos no sul do Brasil. Foram coletadas amostras de três granjas com índice de fetos mumificados ≥ 2,5%. Fragmentos de cérebro, pulmão, rim, fígado e coração foram amostrados para análise por PCR. Das 100 amostras examinadas, 87 foram positivas para PCV2, 68 foram positivas para PPV e 22 foram positivas para Leptospira spp. Além disso, também identificamos co-infecções com dois e três patógenos em uma mesma amostra. Os resultados encontrados contribuem para a disseminação do conhecimento científico relacionado às causas infecciosas (PCV2, PPV e Leptospiras patogênicas) relacionadas às perdas fetais em suínos, assim como evidenciam a alta frequência dos agentes pesquisados.

Perfil de suscetibilidade antimicrobiana e presença do gene vapA em Rhodococcus equi de origem humana, ambiental e equina / Susceptibility profile of Brazilian Rhodococcus equi isolates to different antimicrobial classes and the presence of vapA gene

Rhodococcus equi é um micro-organismo intracelular facultativo, agente etiológico da rodococose, uma importante enfermidade que acomete principalmente potros com menos de seis meses de idade, causando a morte geralmente em decorrência de lesões pulmonares. Este agente também tem potencial zoonótico e emergiu como um patógeno oportunista no mundo, acometendo humanos imunocomprometidos, especialmente os transplantados e infectados pelo vírus da imunodeficiência humana (HIV). Entretanto, infecções por R. equi em hospedeiros hígidos tem sido relatadas, principalmente em crianças e idosos. Estudos tem mostrado um nível crescente na resistência de isolados de R. equi em relação aos antimicrobianos comumente utilizados no tratamento de animais e seres humanos infectados por este agente. A virulência deste pode estar associada a fatores como a cápsula de polissacarídeo, fosfolipase C e à enzima colesterol oxidase (fator equi). No entanto, uma proteína localizada em um plasmídeo, designada vapA, é essencial para a sobrevivência e replicação do agente em macrófagos. Com isso, os objetivos deste estudo foram avaliar o perfil de suscetibilidade de isolados de R. equi de diferentes fontes em relação aos antimicrobianos mais comumente utilizados na terapêutica animal e humana, bem como verificar a associação entre a presença do gene vapA e o índice de resistência múltipla aos antimicrobianos (IRMA). Neste estudo, 67 isolados brasileiros de R. qui de diferentes fontes foram analisados: 30 provenientes de amostras clínicas de equinos, sete de humanos e 30 ambientais (seis do solo e 24 de fezes de equinos). Para avaliar o perfil de suscetibilidade dos isolados utilizou-se o método de disco difusão, sendo testadas 16 drogas de diferentes classes de antimicrobianos. As amostras clínicas de equinos apresentaram as maiores taxas de resistência à penicilina (86,7%) e lincomicina (30%). Além disso, foram também resistentes a macrolídeos (azitromicina a 6,7%, eritromicina a 6% e ...

Rhodococcus equi is a facultative intracellular bacterium and etiological agent of rodococosis, an important disease that affects specially foals under six months old and leading the death generally due to pulmonary lesions. R. equi also has zoonotic potential, and it has emerged as an opportunistic pathogen in the world, specially infecting solid organ transplant recipients and immunocompromised human patients, mainly those infected by human immunodeficiency virus (HIV). Additionally, R. equi infections by healthy hosts have been reported principally in children and elderly individuals. Studies have shown an increasing level of resistance of isolates of R. equi against antibiotics commonly used to treat affected animals and humans. The virulence of this agent is associated with a protein located on a plasmid, designated vapA, it is essential for survival and replication of R. equi within macrophages. The present study evaluated the susceptibility profile of R.equi isolates from different sources against the antimicrobials most commonly used to treat animals and humans, as well as the occurrence and association of vapA gene and the antimicrobial multiple resistance index (AMRI). Sixty-seven Brazilian isolates of R. equi from different sources were analyzed: 30 clinical samples of horses, seven of human and 30 of environmental (six from soil and 24 from horse feces). To evaluate the susceptibility profile of R. equi isolates, the Kirby Bauer method was performed by using 16 drugs of 11 distinct antimicrobials classes. Additionally, those samples were also resistant to macrolides (azithromycin 6.7%, erythromycin 6% and clarithromycin 3.3%) as well as rifamycin (13%). All human and environmental samples were sensitive to macrolides and rifamycin. However, environmental isolates demonstrated high levels of resistance to penicillin and chloramphenicol. Similarly, human isolates had high level of resistance to ceftiofur, lincomycin and sulfazotrim. AMRI...

Phylogenetic and pathotype analysis of Escherichia coli swine isolates from Southern Brazil / Análise filogenética e de patotipos de Escherichia coli isoladas de suínos no Sul do Brazil

The current study evaluated the presence of virulence factors by a multiplex PCR technique and then phylogenetically classified the studied strains into groups A, B1, B2 and D, according to Clermont et al. (2000), in 152 intestinal and extraintestinal swine isolates of Escherichia coli. Seventy seven isolates tested were positive for virulence factors. Phylogenetic characterization placed 21 samples into group A, 65 into B1, 19 into B2 and 47 into D. Fourteen urine samples were classified as uropathogenic E. coli (UPEC), nine were both UPEC and enterotoxigenic E. coli (ETEC) and four were ETEC only. The most common phylogenetic classifications were B1 and D groups. Of the analyzed fecal samples, 25 were classified as ETEC. Phylogenetically, the group of higher occurrence was B1, followed by B2, A and D. For the small intestine samples, 20 were classified as ETEC. Phylogenetic analysis found groups B1 and A to be the most commons in these samples. Six isolated tissue samples were classified as ETEC and most of them were designated as group D by phylogenetic classification. The phylogenetic analysis could be employed in veterinary laboratories in the E. coli isolates screening, including the possibility of vaccine strain selection and epidemiological searches.

O presente estudo teve por objetivo avaliar a presença de diferentes fatores de virulência em 152 isolados de Escherichia coli intestinais e extra-intestinais provenientes de suínos pela técnica de PCR multiplex e classificá-los nos grupos filogenéticos A, B1, B2 e D, de acordo com Clermont et al. (2000). Setenta e sete isolados foram positivos para pelo menos um fator de virulência. Através da caracterização filogenética, 21 isolados foram caracterizados como pertencentes ao grupo A, 65 ao grupo B1, 19 ao grupo B2 e 47 isolados ao grupo D. Quatorze isolados de urina foram caracterizados como E. coli uropatogênica (UPEC); nove apresentaram fatores de UPEC e E. coli enterotoxigênica (ETEC) simultaneamente e quatro foram classificados como ETEC. Na classificação filogenética, os isolados provenientes de amostras de urina classificaram-se principalmente nos grupos D e B1. Das amostras de fezes analisadas, 25 demonstraram fatores de virulência característicos do patotipo ETEC. Filogeneticamente, o grupo de maior ocorrência foi o B1 seguido de B2, A e D. Em relação às cepas isoladas de intestino delgado, 20 foram caracterizadas como ETEC. Pela filogenia, 23 isolados classificaram-se nos grupos A ou B1. Seis isolados de tecidos foram qualificados como ETEC e a maioria deles foram designados como pertencentes ao grupo D, pela classificação filogenética. A análise filogenética pode ser empregada em laboratórios de diagnóstico veterinário como um screening para isolados de E. coli, incluindo a possibilidade de seleção de cepas vacinais e levantamentos epidemiológicos.