ABSTRACT: Bovine mastitis is the most impacting disease of dairy industry, and it is characterized by a complexity of causal agents, which have revealed a geographical variation among regions and countries. The mastitis-related pathogens have been traditionally classified as contagious or environmental, based on habits of the microorganisms and transmission routes. Besides, the severity of mammary infections has been associated with the virulence of the pathogens, and immune and nutritional status of the hosts. Considering this scenario, we investigated the etiological pattern, clinical severity scores, and days in milk (DIM) data in 4,273 clinical cases of bovine mastitis among ten large-dairy farms located in Brazil. Streptococcus dysgalactiae (283/4,273 = 6.6%), Escherichia coli (190/4,273 = 4.4%), Prototheca spp. (112/4,273 = 2.6%), and Streptococcus uberis (95/4,273 = 2.2%) were the predominant pathogens isolated. Among 4,273 clinical cases, clinical severity score was available in 43.8% (1,871/4,273) animals. From these, 69.8% (1,306/1,871), 27.3% (510/1,871), and 2.9% (55/1,871) were scored as mild, moderate, and severe, respectively. Pathogens were observed mainly in samples obtained during the first 100 days in milk, and clinical severity scored mainly as mild. Our results contributed to the etiological identification, clinical severity scoring, and milking aspects of mastitic cows in dairy farms with a history of clinical mammary infections. ABSTRACT industry agents countries mastitisrelated related environmental routes Besides hosts scenario pattern scores DIM (DIM 4273 4 273 4,27 largedairy large Brazil 283/4,273 2834273 283 (283/4,27 6.6%, 66 6.6% , 6 6.6%) 190/4,273 1904273 190 (190/4,27 4.4%, 44 4.4% 4.4%) spp 112/4,273 1124273 112 (112/4,27 2.6%, 26 2.6% 2 2.6%) 95/4,273 954273 95 (95/4,27 2.2% 22 isolated 438 43 8 43.8 1,871/4,273 18714273 1 871 (1,871/4,273 animals these 698 69 69.8 1,306/1,871, 13061871 1,306/1,871 306 (1,306/1,871) 27 3 27.3 510/1,871, 5101871 510/1,871 510 (510/1,871) 29 9 2.9 55/1,871 551871 55 (55/1,871 mild moderate severe respectively 10 identification scoring 427 4,2 283/4,27 283427 28 (283/4,2 6.6 190/4,27 190427 19 (190/4,2 4.4 112/4,27 112427 11 (112/4,2 2.6 95/4,27 95427 (95/4,2 2.2 43. 1,871/4,27 1871427 87 (1,871/4,27 69. 1306187 1,306/1,87 30 (1,306/1,871 27. 510187 510/1,87 51 (510/1,871 2. 55/1,87 55187 5 (55/1,87 42 4, 283/4,2 28342 (283/4, 6. 190/4,2 19042 (190/4, 4. 112/4,2 11242 (112/4, 95/4,2 9542 (95/4, 1,871/4,2 187142 (1,871/4,2 130618 1,306/1,8 (1,306/1,87 51018 510/1,8 (510/1,87 55/1,8 5518 (55/1,8 283/4, 2834 (283/4 190/4, 1904 (190/4 112/4, 1124 (112/4 95/4, 954 (95/4 1,871/4, 18714 (1,871/4, 13061 1,306/1, (1,306/1,8 5101 510/1, (510/1,8 55/1, 551 (55/1, 283/4 (283/ 190/4 (190/ 112/4 (112/ 95/4 (95/ 1,871/4 1871 (1,871/4 1306 1,306/1 (1,306/1, 510/1 (510/1, 55/1 (55/1 283/ (283 190/ (190 112/ (112 95/ (95 1,871/ 187 (1,871/ 130 1,306/ (1,306/1 510/ (510/1 55/ (55/ (28 (19 (11 (9 1,871 18 (1,871 13 1,306 (1,306/ (510/ (55 (2 (1 ( 1,87 (1,87 1,30 (1,306 (510 (5 1,8 (1,8 1,3 (1,30 (51 1, (1, (1,3

RESUMO: A mastite bovina é a doença mais impactante na indústria de laticínios e é caracterizada por uma complexidade de agentes causais, os quais têm revelado uma variação geográfica entre regiões e países. Os patógenos relacionados à mastite têm sido tradicionalmente classificados como contagiosos ou ambientais, com base na fonte de infecção ou origem e nas vias de transmissão. Além disso, a gravidade das infecções mamárias tem sido associada à virulência dos patógenos, e ao estado imunológico e nutricional dos animais. Considerando esse cenário, investigamos os agentes etiológicos, os escores de gravidade clínica e os dados de dias em lactação (DEL) em 4.273 casos clínicos de mastite bovina em dez fazendas de grande porte, quatro localizadas no sudoeste do estado de São Paulo e seis no sul do Estado de Minas Gerais, Brasil. Streptococcus dysgalactiae (283/4.273 = 6,6%), Escherichia coli (190/4.273 = 4,4%), Prototheca spp. (112/4.273 = 2,6%) e Streptococcus uberis (95/4.273 = 2,2%) foram os patógenos predominantes isolados. Entre os 4.273 casos clínicos, a pontuação de gravidade clínica estava disponível em 43,8% (1.871/4.273) dos animais. Desses, 69,8% (1.306/1.871), 27,3% (510/1.871) e 2,9% (55/1.871) foram classificados como leves, moderados e graves, respectivamente. Os patógenos foram observados principalmente em amostras obtidas durante os primeiros 100 dias em lactação, e a gravidade clínica foi principalmente classificada como leve. Nossos resultados contribuem para a identificação etiológica, a pontuação de gravidade clínica e os aspectos de ordenha de vacas com mastite em fazendas leiteiras com histórico de infecções mamárias clínicas. RESUMO causais países ambientais transmissão disso animais cenário etiológicos DEL (DEL 4273 4 273 4.27 porte Gerais Brasil 283/4.273 2834273 283 (283/4.27 6,6%, 66 6,6% , 6 6,6%) 190/4.273 1904273 190 (190/4.27 4,4%, 44 4,4% 4,4%) spp 112/4.273 1124273 112 (112/4.27 2,6% 26 2 95/4.273 954273 95 (95/4.27 2,2% 22 isolados 438 43 8 43,8 1.871/4.273 18714273 1 871 (1.871/4.273 Desses 698 69 69,8 1.306/1.871, 13061871 1.306/1.871 306 (1.306/1.871) 27 3 27,3 510/1.871 5101871 510 (510/1.871 29 9 2,9 55/1.871 551871 55 (55/1.871 leves graves respectivamente 10 leve etiológica clínicas 427 4.2 283/4.27 283427 28 (283/4.2 6,6 190/4.27 190427 19 (190/4.2 4,4 112/4.27 112427 11 (112/4.2 2,6 95/4.27 95427 (95/4.2 2,2 43, 1.871/4.27 1871427 87 (1.871/4.27 69, 1306187 1.306/1.87 30 (1.306/1.871 27, 510/1.87 510187 51 (510/1.87 2, 55/1.87 55187 5 (55/1.87 42 4. 283/4.2 28342 (283/4. 6, 190/4.2 19042 (190/4. 4, 112/4.2 11242 (112/4. 95/4.2 9542 (95/4. 1.871/4.2 187142 (1.871/4.2 130618 1.306/1.8 (1.306/1.87 510/1.8 51018 (510/1.8 55/1.8 5518 (55/1.8 283/4. 2834 (283/4 190/4. 1904 (190/4 112/4. 1124 (112/4 95/4. 954 (95/4 1.871/4. 18714 (1.871/4. 13061 1.306/1. (1.306/1.8 510/1. 5101 (510/1. 55/1. 551 (55/1. 283/4 (283/ 190/4 (190/ 112/4 (112/ 95/4 (95/ 1.871/4 1871 (1.871/4 1306 1.306/1 (1.306/1. 510/1 (510/1 55/1 (55/1 283/ (283 190/ (190 112/ (112 95/ (95 1.871/ 187 (1.871/ 130 1.306/ (1.306/1 510/ (510/ 55/ (55/ (28 (19 (11 (9 1.871 18 (1.871 13 1.306 (1.306/ (510 (55 (2 (1 ( 1.87 (1.87 1.30 (1.306 (51 (5 1.8 (1.8 1.3 (1.30 1. (1. (1.3

Resumen La mastitis bovina es una enfermedad que afecta a las ganaderías de pequeños y medianos productores de los cantones Cayambe y Pedro Moncayo, Provincia de Pichincha-Ecuador. El tratamiento de esta enfermedad resulta complicado debido a la variedad de microorganismos que la provocan. El presente estudio se enfocó en la determinación molecular por medio de reacción en cadena de la polimerasa (PCR) de los agentes etiológicos de la mastitis. Esta técnica presenta múltiples ventajas al reconocer familia, género y especie de microorganismos, y es un método capaz de detectar genes de resistencia de antibióticos, lo que resulta importante al momento de diagnosticar y tratar enfermedades. El objetivo de esta investigación se centró en la identificación de bacterias causantes de la mastitis bovina, utilizando pruebas bioquímicas y moleculares. Las pruebas bioquímicas como tinción Gram, Catalasa, Coagulasa, y Agar Manitol Sal fueron eficientes para obtener cepas puras y determinar el género de algunas bacterias. Se utilizaron primers específicos (RNA16S) para la identificación molecular de 9 agentes etiológicos causantes de la enfermedad en las unidades productivas. Los microorganismos encontrados fueron; Staphylococcus pasteuri, Staphylococcus warneri, Staphylococcus sp., Staphylococcus epidermidis, Staphylococcus aureus, Staphylococcus saprophyticus, Sphingomonas sp., Streptococcus dysgalactiae, Streptococcus uberis, la mayoría presentes en mastitis clínica. Para detectar genes de resistencia se utilizaron primers específicos, de los cuales 7 muestras presentaron el gen para resistencia a blaTEM (β-lactámicos) y 6 muestras presentaron el gen para resistencia a tetA (tetraciclinas). Se identificó multirresistencia en las especies: Staphylococcus pasteuri, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus uberis, Sphingomonas sp.

Abstract Bovine mastitis is a disease that affects the farms of small and medium producers in the cantons of Cayambe and Pedro Moncayo, Pichincha Province-Ecuador. Treating this disease is not easy due to the different microorganisms that cause it. This study focused on the molecular determination by means of polymerase chain reaction (PCR) of the etiological agents of mastitis, having multiple advantages when recognizing family, gender and species of microorganisms. It is a method capable of detecting resistance genes of antibiotics, an important analysis when diagnosing and treating diseases. The aim of this research is to identify bacteria causing bovine mastitis by using biochemical and molecular tests. Biochemical tests such as: Gram staining, Catalase, Coagulase, and Mannitol Salt Agar were efficient to obtain pure strains and determine the gender of some bacteria. Specific primers (RNA16S) were used for the molecular identification of 9 etiological agents causing the disease in the productive units. The microorganisms found were Staphylococcus pasteuri, Staphylococcus warneri, Staphylococcus sp., Staphylococcus epidermidis, Staphylococcus aureus, Staphylococcus saprophyticus, Sphingomonas sp., Streptococcus dysgalactiae, Streptococcus uberis, mostly present in clinic mastitis. To detect resistance genes, specific primers were used, of which 7 samples presented the gene for resistance to blaTEM (β-lactam) and 6 samples presented the gene for resistance to tetA (tetracyclines). Multi-resistance was identified in the species Staphylococcus pasteuri, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus uberis, Sphingomonas sp.

RESUMEN La mastitis linfocítica o mastitis diabética es una patología benigna rara, caracterizada por una inflamación fibrótica de la mama, causando gran preocupación por la simulación de un carcinoma. Está asociada por lo general a la diabetes tipo 2, aunque pueden presentarse en otras patologías autoinmunes y además en pacientes sin comorbilidades. Se presenta en caso clínico de una paciente joven sin comorbilidades que consulto en consultorio de patología mamaria por tumor mamario. Se realizó una biopsia percutánea por trucut con confirmación histológica de mastitis linfocítica, se realizó control cercano observándose remisión clínica e imagenológica de la lesión en 6 meses de seguimiento. Se presenta el caso a fin de recalcar la importancia del diagnóstico histológico percutáneo de la lesión para determinar la naturaliza de la misma, constatándose una patología benigna que no requiere resección, evitando así causar deformidades que podrían afectar psicológicamente a la paciente.

ABSTRACT Lymphocytic mastitis or diabetic mastitis is a rare benign pathology, characterized by fibrotic inflammation of the breast, causing great concern due to the simulation of carcinoma. It is generally associated with type 2 diabetes, although it can occur in other autoimmune pathologies and also in patients without comorbidities. The clinical case of a young patient without comorbidities who consulted in the breast pathology clinic due to a breast tumor. A percutaneous trucut biopsy was performed with histological confirmation of lymphocytic mastitis, close control was performed, observing clinical and imaging remission of the lesion in 6 months of follow-up. The case is presented in order to emphasize the importance of percutaneous histological diagnosis of the lesion, to determine its nature, confirming a benign pathology that does not require resection, thus avoiding causing deformities that could psychologically affect the patient.

Abstract The work aims to analyze the associations of polymorphic variants of the PRL and BLG genes with resistance and susceptibility to mastitis in Holstein cows. The experimental study consisted of the selection of biomaterial samples from 250 heads of Holstein cows aged 3 years divided into two groups (healthy and with a confirmed diagnosis of mastitis). The determination of animal genotypes was carried out using polymerase chain reaction and restriction fragment length polymorphism. The study of the nature of the association of polymorphic variants of the PRL and BLG gene with resistance/increased risk of mastitis established a significant deviation from the theoretically expected distribution of bBLG-HaeIII genotypes in the group of animals suffering from mastitis (the value of χ2 was 0.24). The bBLG-HaeIIIBB genotype can act as a marker of an increased risk of developing mastitis in Holstein cows; its frequency in the group of sick animals exceeds the frequency in the control group by more than 2 times (44.0 compared to 17.0%, respectively). The bBLG-HaeIIIAB genotype is significantly associated with mastitis resistance in Holstein cows; its frequency is 2 times lower than in the control group (28.0 compared to 54.0%). 25 healthy mastitis. . mastitis) polymorphism resistanceincreased bBLGHaeIII bBLG HaeIII χ 0.24. 024 0.24 0 24 0.24) bBLGHaeIIIBB HaeIIIBB 44.0 440 44 (44. 170 17 17.0% respectively. respectively respectively) bBLGHaeIIIAB HaeIIIAB 28.0 280 28 (28. 54.0%. 540 54.0% 54 54.0%) 02 0.2 44. 4 (44 1 17.0 28. (28 54.0 5 0. (4 17. (2 54. (

Resumo O trabalho tem como objetivo analisar as associações de variantes polimórficas dos genes PRL e BLG com resistência e suscetibilidade à mastite em vacas holandesas. O estudo experimental consistiu na seleção de amostras de biomateriais de 250 cabeças de vacas holandesas com 3 anos de idade divididas em dois grupos (saudáveis e com diagnóstico confirmado de mastite). A determinação dos genótipos dos animais foi realizada utilizando reação em cadeia da polimerase e polimorfismo de comprimento de fragmentos de restrição. O estudo da natureza da associação de variantes polimórficas do gene PRL e BLG com resistência/risco aumentado de mastite estabeleceu um desvio significativo da distribuição teoricamente esperada dos genótipos bBLG-HaeIII no grupo de animais que sofrem de mastite (o valor de χ2 foi 0,24). O genótipo bBLG-HaeIIIBB pode atuar como marcador de risco aumentado de desenvolvimento de mastite em vacas holandesas; sua frequência no grupo de animais doentes excede a frequência no grupo controle em mais de duas vezes (44,0% contra 17,0%, respectivamente). O genótipo bBLG-HaeIIIAB está significativamente associado à resistência à mastite em vacas holandesas; sua frequência é duas vezes menor que no grupo controle (28,0% contra 54,0%). 25 saudáveis mastite. . mastite) restrição resistênciarisco bBLGHaeIII bBLG HaeIII o χ 0,24. 024 0,24 0 24 0,24) bBLGHaeIIIBB HaeIIIBB 44,0% 440 44 (44,0 170 17 17,0% respectivamente. respectivamente respectivamente) bBLGHaeIIIAB HaeIIIAB 28,0% 280 28 (28,0 54,0%. 540 54,0% 54 54,0%) 2 02 0,2 44,0 4 (44, 1 17,0 28,0 (28, 54,0 5 0, 44, (44 17, 28, (28 54, (4 (2 (

ABSTRACT: The current study characterizes the genetic distribution of virulence and antimicrobial resistance of Streptococcus lutetiensis and Streptococcus equinus isolated from cows with clinical mastitis using whole genome sequencing (WGS). Although they are not the protagonist species within the genus Streptococcus, recent studies have isolated these species associated with bovine mastitis. In addition, these species are reported and isolated from humans and other animals. A total of four strains of S. lutetiensis and one of S. equinus were isolated from five cows with identified cases of clinical mastitis at a dairy farm near Ithaca, New York. Nineteen genes associated with antimicrobial resistance and 20 genes associated with virulence were identified in the analyzed strains. All strains presented genes associated with resistance: alr, ddl, gdpD, kasA, murA, lsa(E), msr(D), mef(A), gidB, and LiaF. Resistance genes associated with several different classes of antibiotics have also been reported. Sixteen virulence-associated genes were identified in all strains. Based on our findings, we conclude that the studied species have the potential to cause mastitis in cattle, and further studies are important to elucidate their role. ABSTRACT WGS. WGS . (WGS) addition animals S Ithaca York 2 alr ddl gdpD kasA murA lsaE, lsaE lsa E , lsa(E) msrD, msrD msr D msr(D) mefA, mefA mef mef(A) gidB LiaF virulenceassociated findings cattle role (WGS lsa(E msr(D mef(A

RESUMO: O presente estudo caracteriza a distribuição genética de virulência e resistência antimicrobiana de Streptococcus lutetiensis e Streptococcus equinus isolados de vacas com mastite clínica usando sequenciamento completo do genoma. Apesar de não serem as espécies protagonistas dentro do gênero Streptococcus, estudos recentes têm isolado essas espécies associadas à mastite bovina. Além disso, essas espécies são relatadas e isoladas de humanos e outros animais. Um total de quatro cepas de S. lutetiensis e uma de S. equinus foram isoladas de cinco vacas com casos identificados de mastite clínica em uma fazenda leiteira perto de Ithaca, Nova York. Dezenove genes associados à resistência antimicrobiana e 20 genes associados à virulência foram identificados nas cepas analisadas. Todas as linhagens apresentaram genes associados à resistência: alr, ddl, gdpD, kasA, murA, lsa(E), msr(D), mef(A), gidB e LiaF. Genes de resistência associados a várias classes diferentes de antibióticos também foram relatados. Dezesseis genes associados à virulência foram identificados em todas as cepas. Com base em nossos achados, concluímos que as espécies estudadas têm potencial para causar mastite em bovinos e mais estudos são importantes para elucidar seu papel. RESUMO genoma bovina disso animais S Ithaca York 2 analisadas alr ddl gdpD kasA murA lsaE, lsaE lsa E , lsa(E) msrD, msrD msr D msr(D) mefA, mefA mef A mef(A) LiaF relatados achados papel lsa(E msr(D mef(A

ABSTRACT The objective of this study was to evaluate the “in vitro” antimicrobial susceptibility profiles of Staphyloccusspp. and Streptococcus spp. isolated from the milk of cows with infectious mastitis. This study was conducted on 14 dairy farms located in four municipalities on the Island of São Luís, Maranhão, Brazil. The isolates were obtained by bacteriological cultivation of milk samples, followed by morphological and biochemical characterization. Antimicrobial susceptibility tests were performed using the disk diffusion method. The results showed that none of the principles were 100% effective against the isolates. Penicillin, ampicillin, and amoxicillin showed higher rates of resistance in isolates of Coagulase-positive Staphylococcus (80%, 77.2%, and 77.2%, respectively), Coagulase-negative Staphylococcus (80%, 75.4%, and 75.4%, respectively ), and S. aureus (94.3%, 88.6%, and 88.6%, respectively). Isolates of Streptococcus showed a higher frequency of resistance to streptomycin (94%), tetracycline (86%), and lincomycin (76%). Isolates from the Coagulase-negative Staphylococcus group had the highest multidrug resistance profile, with 32.65% of the strains being simultaneously resistant to more than eight antibiotics. A high frequency of isolates of agents that cause bovine mastitis with multidrug-resistant phenotypes was determined, making it necessary to base the treatment of animals on the diagnosis of the causative pathogen and patterns of sensitivity to antimicrobials. vitro Staphyloccusspp spp 1 Luís Maranhão Brazil samples characterization method 100 Penicillin ampicillin Coagulasepositive Coagulase positive 80%, 80 (80% 772 77 2 77.2% respectively, , respectively) Coagulasenegative negative 754 75 4 75.4% ) S 94.3%, 943 94 3 (94.3% 886 88 6 88.6% respectively. . 94%, 94% (94%) 86%, 86 86% (86%) 76%. 76 76% (76%) profile 3265 32 65 32.65 antibiotics multidrugresistant determined antimicrobials 10 80% 8 (80 7 77.2 75.4 94.3% 9 (94.3 88.6 (94% (86% (76% 326 32.6 (8 77. 75. 94.3 (94. 88. (94 (86 (76 32. ( 94. (9 (7

RESUMO Objetivou-se avaliar o perfil de suscetibilidade antimicrobiana “in vitro” de isolados do Staphyloccusspp. e de Streptococcus spp. originados do leite de vacas com mastite infecciosa. O estudo foi realizado em 14 propriedades leiteiras situadas em quatro municípios da Ilha de São Luís, Maranhão. Os isolados foram obtidos do cultivo bacteriológico das amostras de leite seguido de caracterização morfológica e bioquímica. Realizaram-se os testes de suscetibilidade antimicrobiana pela técnica de difusão em discos para 14 princípios antimicrobianos. Os resultados mostraram que nenhum dos princípios apresentou 100% de eficácia contra os isolados. A penicilina, ampicilina e amoxicilina apresentam maiores índices de resistência nos isolados de S. coagulase positiva (80%, 77,2% e 77,2%), S. coagulase negativa (80%, 75,4%, 75,4%) e S. aureus (94,3%, 88,6% e 88,6%), respectivamente. Os isolados do gênero Streptococcus apresentaram maior frequência de resistência a estreptomicina (94%), tetraciclina (86%) e lincomicina (76%). Os isolados do grupo S. coagulase negativa o com maior perfil de multirresistência, com 32,65% das cepas resistentes a mais de oito antibióticos, simultaneamente. Concluiu-se que há alta frequência de isolados de agentes causadores de mastite bovina com fenótipos de multirresistência, sendo necessário embasar o tratamento dos animais no diagnóstico do patógeno causador e nos padrões de sensibilidade a antimicrobianos. Objetivouse Objetivou se in vitro Staphyloccusspp spp infecciosa 1 Luís Maranhão bioquímica Realizaramse Realizaram antimicrobianos 100 penicilina S 80%, 80 (80% 772 77 2 77,2 77,2%, , 77,2%) 754 75 4 75,4% 94,3%, 943 94 3 (94,3% 886 88 6 88,6 88,6%, 88,6%) respectivamente 94%, 94% (94%) 86% 86 (86% 76%. 76 76% . (76%) multirresistência 3265 32 65 32,65 antibióticos simultaneamente Concluiuse Concluiu 10 80% 8 (80 7 77, 75,4 94,3% 9 (94,3 88, (94% (86 (76% 326 32,6 (8 75, 94,3 (94, (94 (76 32, ( 94, (9 (7

ABSTRACT This study aimed to assess the antimicrobial susceptibility of 52 Staphylococcus aureus (S. aureus) bovine mastitis isolates obtained from 37 dairy herds from the northwest of Portugal against antibiotics belonging to the β-lactam family, evaluate the detection of blaZ and mecA resistance genes, and study the diversity of positive isolates. The antimicrobial susceptibility tests were performed by the disk diffusion method. The detection of blaZ and mecA genes was performed using specific polymerase chain reaction (PCR) and the diversity of blaZ was evaluated by phylogenetic analysis. The antimicrobial susceptibility test showed a prevalence of phenotypic resistance by S. aureus of 52.0% against penicillin and ampicillin, 34.6% against oxacillin, 17.3% against amoxicillin plus clavulanic acid, and 21.1% against cefazolin. A prevalence of phenotypic intermediate resistance of 5.7% against penicillin, 9.6% against amoxicillin plus clavulanic acid, and 1.9% against ampicillin and cefazolin, respectively, was demonstrated. A 100.0% phenotypic susceptibility was found against piperacillin. Of the 52 S. aureus isolates, 35 (67.3%) were PCR positive for blaZ gene, and isolate 25 was positive for mecA gene. The phylogenetic analysis of partial blaZ gene S. aureus isolates consensus sequences were placed in two different clades, clade A (cluster A, A.1) and B (cluster B), all closely related to animal and/or human S. aureus strains. Isolate 2 appeared in the phylogenetic tree as the most divergent. This study indicates that blaZ resistance gene plays a role in β-lactam resistance in the tested bovine mastitis S. aureus isolates within dairy herds in the northwest of Portugal, especially in case of penicillin and ampicillin antibiotics that have shown a high phenotypic prevalence. Indeed, the proportion of bovine mastitis isolates with phenotypic resistance did not agree with the proportion of those identified for blaZ, as isolates with 100.0% of phenotypic susceptibility for all tested antibiotics also harbored blaZ. BlaZ phylogenetic analysis from S. aureus isolates showed diversity inside or between different herds in the northwest of Portugal. Piperacillin, as a suggestion, could be tested for S. aureus bovine mastitis treatment in the future to evaluate this new possibility of therapy. 5 S (S 3 βlactam β lactam family method (PCR 520 0 52.0 346 34 6 34.6 oxacillin 173 17 17.3 acid 211 21 1 21.1 cefazolin 57 7 5.7 96 9 9.6 19 1.9 respectively demonstrated 1000 100 100.0 piperacillin 67.3% 673 67 (67.3% clades cluster A.1 A1 B, , B) andor strains divergent Indeed Piperacillin suggestion therapy 52. 34. 17. 21. 5. 9. 1. 10 100. 67.3 (67.3 A. 67. (67. (67 (6 (

ABSTRACT: Escherichia coli is recognized as one of the main microorganisms responsible for triggering clinical mastitis, a disease that causes considerable economic losses in the dairy industry. In this context, this study aimed to identify E. coli isolates present in individual milk samples collected from cows diagnosed with clinical mastitis from various regions of Brazil. Additionally, through polymerase chain reaction (PCR), the presence of virulence genes eae, bfpB, escN, aatA, aggR, ipaH, stx1, stx2, est, and eltA was investigated; all associated with the pathotypes of diarrheagenic Escherichia coli (DEC). As an integral part of the study, a comprehensive assessment of the sensitivity profile of the isolates to 11 different antimicrobials widely used in mastitis treatment was also conducted. A total of 198 milk samples were collected from cows diagnosed with clinical mastitis. Among these samples, 12 isolates (6.07%) demonstrated bacterial growth greater than three Colony-Forming Units (CFU) when grown on MacConkey agar medium and morphological characteristics of E. coli. The disc-diffusion test was used to evaluate the susceptibility of these isolates to antimicrobials, and the most predominant resistance was observed concerning streptomycin and tetracycline, affecting 16.67% of the strains analyzed. Notably, all isolates investigated did not demonstrate the presence of the genes eae, aatA, aggR, ipaH, stx1, stx2, est, and eltA. These results indicate that these isolates do not fit the pathotypes known as diarrheagenic Escherichia coli (DEC). However, one of the isolates tested was positive for the bfpB and escN genes. The detection of resistant E. coli associated with clinical mastitis points to possible gaps in the treatment of the disease. Additionally, the presence of resistance genes in E. coli strains indicates the potential to transmit these genes between animals and, perhaps, along the food chain. ABSTRACT industry context E Brazil Additionally PCR, PCR , (PCR) eae aatA aggR ipaH stx1 stx stx2 est DEC. DEC . (DEC) 1 conducted 19 6.07% 607 6 07 (6.07% ColonyForming Colony Forming CFU (CFU discdiffusion disc diffusion tetracycline 1667 16 67 16.67 analyzed Notably However perhaps (PCR (DEC 6.07 60 0 (6.07 166 16.6 6.0 (6.0 16. 6. (6. (6 (

RESUMO: Escherichia coli é reconhecida como um dos principais microrganismos responsáveis pelo desencadeamento da mastite clínica, doença que causa perdas econômicas consideráveis na indústria de laticínios. Neste contexto, este estudo teve como objetivo principal a identificação de isolados de E. coli presentes em amostras individuais de leite coletadas de vacas com diagnóstico de mastite clínica, de diversas regiões do Brasil. Adicionalmente, por reação em cadeia da polimerase (PCR), foi investigada a presença dos genes de virulência eae, bfpB, escN, aatA, aggR, ipaH, stx1, stx2, est e eltA, todos associados aos patótipos de E. coli diarreiogênica (DEC). Como parte integrante do estudo, foi realizada uma avaliação abrangente do perfil de sensibilidade dos isolados a 11 antimicrobianos diferentes amplamente utilizados no tratamento da mastite. Foram coletadas 198 amostras de leite de vacas com diagnóstico de mastite clínica. Dentre essas amostras, 12 isolados (6,07%) demonstraram crescimento bacteriano superior a três Unidades Formadoras de Colônia (UFC) quando cultivadas em meio ágar MacConkey e características morfológicas de E. coli. Para avaliar a suscetibilidade desses isolados aos antimicrobianos foi utilizado o teste de disco-difusão, sendo observada a resistência mais predominante em relação à estreptomicina e à tetraciclina, afetando 16,67% das cepas analisadas. É relevante ressaltar que todos os isolados investigados não demonstraram a presença dos genes eae, aatA, aggR, ipaH, stx1, stx2, est e eltA. Estes resultados indicam que estes isolados não se enquadram nos patótipos conhecidos como Escherichia coli diarreiogénica (DEC). Porém, um dos isolados testados apresentou positividade para os genes bfpB e escN. A detecção de E. coli resistente associada à mastite clínica aponta para possíveis lacunas no tratamento da doença. Além disso, a presença de genes de resistência em estirpes de E. coli indica a capacidade potencial de transmitir estes genes entre animais e talvez ao longo da cadeia alimentar. RESUMO laticínios contexto E Brasil Adicionalmente PCR, PCR , (PCR) eae escN aatA aggR ipaH stx1 stx stx2 eltA DEC. DEC . (DEC) 1 19 6,07% 607 6 07 (6,07% UFC (UFC discodifusão, discodifusão disco difusão, difusão disco-difusão tetraciclina 1667 16 67 16,67 analisadas Porém disso alimentar (PCR (DEC 6,07 60 0 (6,07 166 16,6 6,0 (6,0 16, 6, (6, (6 (

ABSTRACT. The study was carried out in the region of Placas, Pará. Objective: identify the main causing agents of mastitis, as well as evaluating the resistance profile of the isolated agents against antimicrobials. Methods: Ten properties were evaluated, where 161 animals were submitted to the California Mastitis Test (CMT) for the diagnosis of subclinical mastitis, and milk samples were collected from positive animals and led for culture and antibiogram. Results: In total, 31.06% of the animals were positive for CMT. The main agents identified were Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus agalactiae and Streptococcus bovis. The antibiogram indicated resistance to Penicillin, Ceftazidime and Cephalothin. Conclusion: The properties surveyed showed a high rate of subclinical mastitis, regarding antimicrobial resistance, in general, all agents showed high resistance. ABSTRACT Placas Pará Objective mastitis antimicrobials Methods evaluated 16 CMT (CMT Results total 3106 31 06 31.06 aureus epidermidis bovis Penicillin Cephalothin Conclusion general 1 310 3 0 31.0 31.

ABSTRACT: The association of amoxicillin and clavulanic acid has shown high efficacy to treat mastitis worldwide, also promoting a shorter milk disposal period compared to other antimicrobials. However, no previous investigations regarding its application to treat mastitis in Brazilian dairy cows were developed. This study evaluated the effectiveness of amoxicillin-clavulanic acid to treat Brazilian dairy cows with mastitis, comparing it with a reference protocol treatment combination of tetracycline, neomycin and bacitracin. Holstein cows (n = 468) from three different dairy herds in Brazil were assigned to two groups: combination of tetracycline, neomycin and bacitracin (n = 178; positive control group) and amoxicillin-clavulanic acid protocol (n = 290). Before the treatment, milk samples were collected and cultivated in chromogenic media. After finishing the antimicrobial treatment (intramammary), milk samples were also collected and cultivated in chromogenic media. Results from microbiological analysis obtained before and after treatment were compared to determine the healing rate. Nine different microbiological agents were identified: eight of environmental origin and one of contagious origin; being eight gram-positive and one gram-negative bacteria. TThe positive control group and the treatment group showed similar healing rate (86.5% and 90.3%, respectively; P > 0.05). No differences were found between the groups for the healing rate, when the causing agent was considered. Therefore, it is possible to indicate the amoxicillin-clavulanic acid-based protocol to treat intramammary mastitis in Brazilian dairy cows, achieving great healing rates and providing a substantial reduction in milk disposal. ABSTRACT worldwide antimicrobials However developed amoxicillinclavulanic tetracycline n 468 178 290. 290 . 290) media intramammary, , (intramammary) identified grampositive gram gramnegative negative bacteria 86.5% 865 86 5 (86.5 903 90 3 90.3% respectively 0.05. 005 0.05 0 05 0.05) considered Therefore acidbased based 46 17 29 (intramammary 86.5 8 (86. 9 90.3 00 0.0 4 1 2 86. (86 90. 0. (8 (

RESUMO: A associação de amoxicilina e ácido clavulânico tem mostrado alta eficácia para o tratamento mastite em diversos países, além de promover um menor período de carência em comparação a outros antimicrobianos utilizados no tratamento da mastite. Apesar disso, estudos considerando a sua utilização para o tratamento de mastite bovina nos rebanhos leiteiros brasileiros ainda não foram realizados. Portanto, esse estudo teve o objetivo de avaliar a efetividade do uso de amoxicilina associado ao ácido clavulânico no tratamento da mastite bovina em rebanhos leiteiros do Brasil e compará-los com os resultados obtidos a partir do uso de um protocolo referência (contendo tetraciclina, neomicina e bacitracina). Um total de 468 vacas da raça Holandesa de três diferentes rebanhos brasileiros foi utilizado no estudo, sendo 178 animais designados para o grupo tetraciclina, neomicina e bacitracina (controle positivo) e 290 animais designados para o grupo amoxicilina-ácido clavulânico. Antes de iniciar o tratamento, amostras de leite foram coletadas e cultivadas em meio cromogênico. Após finalizar o tratamento com os antimicrobianos, por via intramamária, amostras de leite foram novamente coletadas e cultivadas em meio cromogênico. Os resultados das análises microbiológicas realizadas antes e após a administração foram comparados para calcular a taxa de cura dos tratamentos. Nove diferentes agentes foram identificados nas amostras de leite (oito de origem ambiental e um de origem contagiosa, sendo oito agentes gram-positivos e um gram-negativo). O grupo de vacas tratadas com amoxicilina-ácido clavulânico apresentou taxa de cura semelhante à taxa de cura dos animais do grupo controle positivo (90,3% vs. 86,5%; P > 0.05). A taxa de cura dos grupos foi semelhante também quando se considerou o agente etiológico. Portanto, é possível indicar a associação de amoxicilina com ácido clavulânico no tratamento da mastite bovina em rebanhos brasileiros, atingindo boas taxas de cura e possibilitando expressiva redução no período de descarte do leite. RESUMO países disso realizados Portanto comparálos compará los contendo tetraciclina bacitracina. . bacitracina) 46 17 29 amoxicilinaácido cromogênico intramamária tratamentos contagiosa grampositivos gram positivos gramnegativo. gramnegativo negativo gram-negativo) 90,3% 903 90 3 (90,3 vs 86,5% 865 86 5 0.05. 005 0.05 0 05 0.05) etiológico 4 1 2 gram-negativo 90,3 9 (90, 86,5 8 00 0.0 90, (90 86, 0. (9 (

Abstract The aim of the study was to evaluate the use of Acustic Radiation Force Impulse (ARFI) elastography in mammary parenchyma and supramammary lymph nodes, for detection of active mastitis in sheep with naturally infected chronic fibrous lesions. 27 female sheep were included and B-mode ultrasound and ARFI elastography images were obtained, acquiring qualitative (echogenicity and echotexture) and quantitative (shear rate, depth and short/long axis ratio) variables of 48 mammary glands. The glands were divided into three experimental groups: control group (CG) - healthy animals; LSCC- animals that presented fibrous lesions and SCC (somatic cell count) less than 500 x 103 cls/mL; HSCC: animals that presented fibrous lesions and SCC (somatic cell count) more than 500 x 103 cls/mL; The qualitative variables using B-mode ultrasonography, including echotexture and echogenicity, showed no significant differences between the evaluated groups and tissues (p = 0.9336 and p = 0.233, respectively) .In healthy areas of the gland, it was an increasing in shear wave velocity (SWV) in LSCC than in HSCC (p=0.04). When comparing the fibrosis in the LSCC and HSCC groups with their respective normal areas, the velocity increased in both groups: LSCC (p= 0,0007) and HSCC (p= 0,0001). When comparing the areas of fibrosis in LSCC and HSCC with the CG parenchyma, there was an increase in LSCC (p=0.001) and HSCC (p=0.0001). B-mode ultrasound indicate predominance of hypoechoic echogenicity in lymph nodes and reduced short/long axis ratio in cases of active subclinical mastitis. The supramammary lymph node showed increased SWV when comparing the CG with HSCC groups (p=0.02) and GC with LSCC (p=0.04). B-mode ultrasonography is useful for evaluating the mammary parenchyma, however, its application as a standalone diagnostic technique is not recommended. ARFI elastography indicates potential cutoff points for differentiating subclinical mastitis from healed mastitis, highlighting its importance as a tool for distinguishing normal areas from fibrous parenchymal areas. While this study did not establish specific cutoff points due to sample size limitations, further research with larger sample sizes could explore and define these critical thresholds. (ARFI 2 Bmode B mode obtained rate shortlong short long 4 (CG somatic count 50 10 cls/mL clsmL cls mL 09336 0 9336 0.933 0233 233 0.233 respectively In gland (SWV p=0.04. p004 p=0.04 . 04 (p=0.04) p= 0,0007 00007 0007 0,0001. 00001 0,0001 0001 0,0001) p=0.001 p0001 001 (p=0.001 p=0.0001. p00001 p=0.0001 (p=0.0001) p=0.02 p002 02 (p=0.02 however recommended limitations thresholds 5 1 0933 933 0.93 023 23 0.23 p00 p=0.0 (p=0.04 0,000 0000 000 p=0.00 p000 00 (p=0.00 p0000 p=0.000 (p=0.0001 (p=0.0 093 93 0.9 0.2 p0 p=0. 0,00 (p=0.000 (p=0. 09 9 0. p=0 0,0 (p=0 0,

ABSTRACT Tuberculous mastitis (TM) is a rare cause of breast disease. The test sensitivity for TM is low and sometimes characterized by idiopathic granulomatous mastitis. Both have different treatments; however, many patients are treated with presumptive TM (P-TM). We present fifteen women with confirmed TM or P-TM. We found 4% (5/124) of TM and 8% (10/124) of P-TM, grouped according to their diagnosis, with no differences between them, except for follow-up and treatment time. P-TM is an entity that must be considered based on clinical and radiological suspicion and is associated with presumptive pathological data. (TM disease treatments however PTM. PTM P . (P-TM) TM. 4 5/124 5124 5 124 (5/124 8 10/124 10124 10 (10/124 PTM, TM, diagnosis them followup follow up time data (P-TM 5/12 512 12 (5/12 10/12 1012 1 (10/12 5/1 51 (5/1 10/1 101 (10/1 5/ (5/ 10/ (10/ (5 (10 ( (1

RESUMO O objetivo do presente ensaio clínico foi avaliar o consumo de matéria seca (CMS) e o comportamento de vacas diagnosticadas com mastite clínica (MC) leve ou moderada, provocada por patógenos Gram negativos ou com cultura negativa. Os casos foram designados para o grupo controle negativo ou grupo tratamento, o qual recebeu uma infusão intramamária de ciprofloxacino por três dias consecutivos. O comportamento foi avaliado por meio de coleiras que monitoram os tempos de ruminação, de atividade e de ócio. Cochos eletrônicos foram utilizados para registrar o comportamento alimentar e o CMS. Além disso, a produção média diária de leite, a composição do leite, a contagem de células somáticas e a contagem de leucócitos também foram avaliadas. A partir de 67 casos de MC, 23 atenderam aos critérios de inclusão e foram designados para o grupo controle negativo (n=13) e para o grupo tratamento (n=10). Não foram encontradas diferenças no comportamento entre os grupos, durante o período experimental. Ademais, não houve diferença para o CMS, o comportamento alimentar e os demais parâmetros, entre o controle negativo e os casos tratados. Assim, o tratamento seletivo não trouxe impactos negativos sobre os parâmetros estudados. Essa estratégia terapêutica pode ser benéfica por reduzir custos e o uso de antimicrobianos, nas propriedades leiteiras. CMS (CMS MC (MC moderada negativa consecutivos ruminação ócio disso leite avaliadas 6 2 n=13 n13 n 13 (n=13 n=10. n10 n=10 . 10 (n=10) grupos experimental Ademais tratados Assim estudados antimicrobianos leiteiras n=1 n1 1 (n=1 (n=10 n= (n= (n

ABSTRACT The aim of the present negatively controlled, randomized clinical trial was to assess dry matter intake (DMI) and behavior in cows with mild and moderate naturally occurring CM caused by gram-negative or culture-negative pathogens. Enrolled cases were assigned to either the negative control or to the treatment group, which received one infusion of intramammary ciprofloxacin for three consecutive days. Behavior was assessed through neck collars which monitors rumination, activity, and rest times. Electronic feeders were used to record feeding behavior and DMI. Also, average daily milk yield, milk composition, somatic cells count (SCC), and white blood cells count (WBC) were evaluated. Of a total of 67 cases of CM, 23 met the criteria to enter the experiment and were enrolled to the negative control (n=13) and to the treated group (n=10). There was no difference in behavior between groups throughout the studied period. Also, no difference was found in DMI, feeding behavior and in the other parameters between negative control and treated cases. Therefore, the selective treatment did not have negative impacts on behavior and DMI, as well as in the other parameters. This therapeutic strategy can be beneficial since reduces costs and the use of antimicrobials in dairy farms. controlled DMI (DMI gramnegative gram culturenegative culture pathogens days rumination activity times Also yield composition SCC, SCC , (SCC) WBC (WBC evaluated 6 2 n=13 n13 n 13 (n=13 n=10. n10 n=10 . 10 (n=10) period Therefore farms (SCC n=1 n1 1 (n=1 (n=10 n= (n= (n

Abstract Brazil is a large sheep and cow producer, and the development of new strategies for the treatment of lamb umbilical cord healing as well as the bovine mastitis is very important. The aim of this study was to develop emulgels containing propolis, curcumin and andiroba oil for the treatment of mastitis and umbilical cord healing in animals. The formulations were composed of poly (acrylic acid) derivative (Carbopol 974P or polycarbophil), Brazilian green propolis extract, curcumin, and andiroba oil. The systems were investigated as the activity on umbilical cord healing of newborn lambs, and evaluation of the prevention of mastitis in dairy cows. The emulgels effectively promoted umbilical cord healing, preventing infections. Moreover, they were effective for reducing the somatic cell count a health indicator for mammary glands in cows. These findings suggest that these formulations can potentially treat cattle mastitis and improve lambs' umbilical cord healing. producer important animals acrylic acid Carbopol P polycarbophil, polycarbophil , polycarbophil) extract lambs cows infections Moreover

ABSTRACT. Four strains of Staphylococcus aureus isolated from cows with mastitis were assessed as to in vitro sensitivity to four commercial disinfectants (chlorine, polvidone-iodine, chlorhexidine, and lactic acid) used in teat pre- and post-dipping, in the presence and absence of organic matter. The concentration of the disinfectants was determined in accordance with the Brazilian Pharmacopoeia. Four isolates identified as S. aureus from cows diagnosed with subclinical mastitis were subjected to susceptibility testing against disinfectants through an absorbance evaluation at 600 nm on a spectrophotometer. Exposure times (15”, 30”, 60” and 90”) were clocked, and streaking was performed in Brain Heart Infusion (BHI) medium. The iodine sample presented a lower concentration (1.9%) than that given by the manufacturer (2.5%). There was greater (p < 0.05) in vitro disinfectant activity for chlorhexidine and chlorine, with and without organic matter. However, inhibition by organic matter occurred more intensely in the iodine disinfectant (99.0%). The inhibition percentage of S. aureus against iodine and lactic acid at 15, 30, 60 and 90 s was numerically lower than against chlorhexidine and chlorine, regardless of organic matter. Therefore, chlorhexidine and chlorine have greater inhibitory activity against S. aureus, both with and without organic matter. ABSTRACT (chlorine polvidoneiodine, polvidoneiodine polvidone iodine, polvidone-iodine pre postdipping, postdipping post dipping, dipping post-dipping Pharmacopoeia S spectrophotometer 15 , (15” 30 30” 90” clocked BHI (BHI medium 1.9% 19 1 9 (1.9% 2.5%. 25 2.5% . 2 5 (2.5%) p 0.05 005 0 05 However 99.0%. 990 99.0% 99 (99.0%) 6 Therefore (15 3 1.9 (1.9 2.5 (2.5% 0.0 00 99.0 (99.0% (1 1. (1. 2. (2.5 0. 99. (99.0 ( (2. (99. (2 (99 (9