Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 855
Filtrar
1.
Int J Food Microbiol ; 334: 108828, 2020 Dec 02.
Artigo em Inglês | MEDLINE | ID: mdl-32866940

RESUMO

Raw milk is a continued threat to public health due to possible contamination with zoonotic pathogens. Enterocytozoon bieneusi is one of the most prevalent pathogenic fungi in a wide range of vertebrate hosts, causing diarrheal disease. Although there has been some evidence, the role and potential risk of raw milk of dairy animals in the transmission dynamics of E. bieneusi is not clear. Therefore, we aimed to determine the occurrence and genotypes of E. bieneusi in raw milk of dairy animals in several farms of the Central Anatolia Region. We also investigated if there is a relation between the presence of E. bieneusi and mastitis. Genomic DNAs from a total of 450 raw milk including 200, 200 and 50 samples from cattle, sheep and water buffalo respectively were analyzed using nested PCR, targeting the internal transcribed spacer of E. bieneusi. Totally milk samples of 9 (4.5%) dairy cattle, 36 (18.0%) sheep, and 1 (2.0%) water buffalo were PCR-positive. A significant relationship was determined between mastitis and the presence of E. bieneusi. Sequence analysis revealed the presence of eight genotypes: two known (ERUSS1, BEB6) and six novel genotypes (named as TREb1 to TREb6). The genotype ERUSS1 and BEB6 were the most common genotypes, found in all cattle and sheep farms. Phylogenetic analysis clustered all the identified genotypes in Group 2. This study provides novel findings that contribute to the transmission dynamics and molecular epidemiology of E. bieneusi. Our study also highlighted the potential risk of raw milk for public health with respect to microsporidia infections.


Assuntos
Doenças dos Bovinos/epidemiologia , Enterocytozoon/genética , Microsporidiose/veterinária , Leite/microbiologia , Doenças dos Ovinos/epidemiologia , Animais , Búfalos , Bovinos , Doenças dos Bovinos/microbiologia , Doenças dos Bovinos/transmissão , Enterocytozoon/classificação , Enterocytozoon/isolamento & purificação , Fazendas , Feminino , Genótipo , Mastite/epidemiologia , Mastite/microbiologia , Mastite/veterinária , Microsporidiose/epidemiologia , Microsporidiose/microbiologia , Microsporidiose/transmissão , Epidemiologia Molecular , Filogenia , Prevalência , Ovinos , Doenças dos Ovinos/microbiologia , Doenças dos Ovinos/transmissão , Turquia
2.
PLoS One ; 15(5): e0231168, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32365127

RESUMO

Staphylococcus aureus is one of the most important pathogens causing mastitis in dairy cows. The objective of this study was to establish a rat model of mastitis induced by S. aureus infection and to explore changes in the proteomes of mammary tissue in different udder states, providing a better understanding of the host immune response to S. aureus mastitis. On day 3 post-partum, 6 rats were randomly divided into two groups (n = 3), with either 100 µL of PBS (blank group) or a S. aureus suspension containing 2×107 CFU·mL-1 (challenge group) infused into the mammary gland duct. After 24 h of infection, the rats were sacrificed, and mammary gland tissue was collected. Tandem mass tag (TMT)-based technology was applied to compare the proteomes of healthy and mastitic mammary tissues. Compared with the control group, the challenge group had 555 proteins with significant differences in expression, of which 428 were significantly upregulated (FC>1.2 and p<0.05) and 127 were downregulated (FC>0.83 and p<0.05 or p<0.01). Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses revealed that upregulated differentially significant expressed proteins (DSEPs) were associated with mainly immune responses, including integrin alpha M, inter-α-trypsin inhibitor heavy chain 4, and alpha-2-macroglobulin. This study is the first in which a rat model of S. aureus-induced mastitis was used to explore the proteins related to mastitis in dairy cows by TMT technology, providing a model for replication of dairy cow S. aureus-induced mastitis experiments.


Assuntos
Glândulas Mamárias Animais/metabolismo , Mastite/metabolismo , Proteoma/análise , Infecções Estafilocócicas/metabolismo , Staphylococcus aureus/fisiologia , Animais , Cromatografia Líquida de Alta Pressão , Cromatografia Líquida , Feminino , Glândulas Mamárias Animais/microbiologia , Glândulas Mamárias Animais/patologia , Mastite/microbiologia , Mastite/patologia , Gravidez , Proteoma/metabolismo , Proteômica/métodos , Ratos , Ratos Sprague-Dawley , Infecções Estafilocócicas/patologia , Espectrometria de Massas em Tandem
3.
PLoS One ; 15(5): e0233284, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32469943

RESUMO

Human milk provides complete nutrition for infants and at the same time promotes the growth of specific bacteria in the infant gastrointestinal tract. Breastfeeding can often be discontinued due to mastitis which is an inflammation of the breast tissue. We isolated 18 Staphylococcus aureus strains from milk donated by healthy (n = 6), subclinical (n = 6), and mastitic (n = 6) mothers, two strains of which were VISA (Vancomycin Intermediate S. aureus). All tested strains (n = 12) were able to form biofilms. We then examined the impact of nisin A and vancomycin alone and in combination on biofilm formation and eradication of selected strains (n = 8). We observed strain-specific responses, with the combinatorial treatment at 1/4X MIC (for both singularly) significantly inhibiting biofilm formation for seven out of eight strains when compared with nisin A or vancomycin alone. None of the selected treatments were able to eradicate pre-formed biofilms. Finally, we selected two strains, namely a VISA (APC3814H) and a strong biofilm former (APC3912CM) and used confocal microscopy to evaluate the effects of the antimicrobial agents at 1X MIC on biofilm inhibition and eradication. All treatments inhibited biofilm formation of APC3814H but were ineffective in eradicating a pre-formed biofilm. Single treatments at 1X MIC against APC3912CM cells did not prevent biofilm formation whereas combination treatment caused increased death of APC3912CM cells. Finally, the combination treatment reduced the thickness of the pre-formed APC3912CM biofilm as compared with the single treatments.


Assuntos
Biofilmes/efeitos dos fármacos , Mastite/tratamento farmacológico , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Leite Humano/efeitos dos fármacos , Nisina/uso terapêutico , Infecções Estafilocócicas/tratamento farmacológico , Vancomicina/uso terapêutico , Antibacterianos/uso terapêutico , Biofilmes/crescimento & desenvolvimento , Quimioterapia Combinada , Feminino , Humanos , Mastite/microbiologia , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Testes de Sensibilidade Microbiana , Leite Humano/microbiologia , Infecções Estafilocócicas/microbiologia
4.
J Dairy Sci ; 103(5): 4717-4731, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32171518

RESUMO

Mastitis is an important constraint to milk production in pastoralist camel (Camelus dromedarius) herds in Kenya. The objective of this study was to investigate the prevalence, risk factors, and bacterial panorama of subclinical mastitis (SCM) in pastoralist camel herds in Isiolo County, Kenya. Furthermore, antimicrobial susceptibility in udder pathogens was studied. A cross-sectional sample of 206 camels from 20 milking herds was screened using the California Mastitis Test (CMT), and quarter milk was subjected to bacterial culturing. Isolates were confirmed using MALDI-TOF mass spectrometry analysis, and antimicrobial susceptibility was determined using the broth microdilution method. Interviews focusing on herd management were conducted with camel owners. Subclinical mastitis, defined as a CMT score ≥ 3 (scale 1 to 5) and absence of clinical symptoms in the udder, were present in all visited herds. On the individual level, 46% of the camels had at least 1 quarter affected with SCM, and on the quarter level the prevalence was 26%. Intramammary infections (IMI) were common; out of 798 quarter milk samples, 33% yielded conclusive bacterial growth. The sensitivity and specificity of CMT for correctly identifying quarters with IMI were 82% and 92%, respectively. The most prevalent pathogen was Streptococcus agalactiae (72% of IMI-positive quarters), followed by non-aureus staphylococci (19%) and Staphylococcus aureus (13%). Antimicrobial susceptibility testing revealed that only a low proportion (4.9%) of Strep. agalactiae isolates was sensitive to tetracycline. For Staph. aureus, 59.1% of isolates exhibited sensitivity to penicillin. Skin lesions on the teats or udder were a risk factor for SCM. Increased age, parity, and stage of lactation were associated with increased risk of both SCM and IMI. Older camels with a blind teat or a previous history of mastitis were more likely to be infected with Strep. agalactiae. Hygiene routines for milking were largely absent in the observed herds, and knowledge of adequate milk handling was limited. The poor udder health is likely to depend on multiple factors, most prominently the within-herd maintenance of contagious udder pathogens, in combination with difficult sanitary conditions and lack of awareness among camel keepers. This study showed that in pastoralist camel herds around Isiolo town, SCM and IMI specifically caused by Strep. agalactiae are common udder health problems and are associated with increasing age, parity, and stage of lactation, and skin lesions on the teats and udder. Resistance to tetracycline in Strep. agalactiae was common. Control strategies specifically targeting SCM and adapted to pastorally managed camel herds need to be developed to reduce disease, combat antimicrobial resistance, and improve the livelihoods of pastoralists.


Assuntos
Antibacterianos/farmacologia , Camelus/microbiologia , Farmacorresistência Bacteriana , Mastite/veterinária , Leite/microbiologia , Infecções Estafilocócicas/veterinária , Streptococcus/classificação , Animais , Estudos Transversais , Feminino , Geografia , Higiene , Quênia/epidemiologia , Lactação , Glândulas Mamárias Animais/microbiologia , Mastite/epidemiologia , Mastite/microbiologia , Leite/metabolismo , Prevalência , Fatores de Risco , Infecções Estafilocócicas/epidemiologia , Infecções Estafilocócicas/microbiologia , Staphylococcus aureus/classificação , Streptococcus agalactiae/classificação , Tetraciclina/farmacologia
5.
J Dairy Sci ; 103(4): 3648-3655, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32089296

RESUMO

In dairy cattle, mastitis is a disease of the mammary gland caused by pathogens such as bacteria, viruses, fungi, and algae. Mastitis causes economic losses to dairy farms as well as public health concerns. The reproductive efficiency of commercial dairy herds has important implications for the economic success of dairy operations and is strongly associated with the health status of cows. Mastitis has previously been linked with decreased fertility of dairy cows, but the effect of specific pathogens on the severity of fertility reduction is still unclear. In this study, cows diagnosed with mastitis caused by major pathogens (Staphylococcus aureus, Streptococcus agalactiae, Escherichia coli, Klebsiella spp., Mycoplasma spp., and environmental Streptococcus) needed more artificial inseminations (AI) than did cows with mastitis caused by minor pathogens (coagulase-negative Staphylococcus and Corynebacterium spp.) and healthy cows. Cows diagnosed with mastitis, independent of what pathogen was causing mastitis, had more days open compared with nonmastitic cows. The percentage of cows that successfully established pregnancy at first AI was greater for the control group than for the major pathogens group but not significantly different from the minor pathogens group. Pregnancy loss was lower in the control group than in the major pathogens group; however, there was no difference compared with the minor pathogen group. Mastitis caused by gram-negative bacteria decreased the percentage of pregnancy per first AI and increased days open and pregnancy loss compared with the control group. Cows with mastitis caused by gram-positive bacteria also had increased days open compared with control cows. This study shows that different mastitis-causing bacteria can affect the fertility of cows differently. Mastitis events caused by major pathogens and gram-negative bacteria were associated with the greatest decrease in reproductive efficiency.


Assuntos
Fenômenos Fisiológicos Bacterianos , Interações entre Hospedeiro e Microrganismos/fisiologia , Inseminação Artificial/veterinária , Mastite Bovina/microbiologia , Mastite/veterinária , Reprodução , Animais , Bactérias/classificação , Bovinos , Feminino , Inseminação Artificial/estatística & dados numéricos , Mastite/microbiologia , Leite/microbiologia
6.
Biomed Pharmacother ; 123: 109779, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31918211

RESUMO

Staphylococcus xylosus (S. xylosus) is one of the emerging pathogens causing bovine mastitis with high rate of isolation in most of the reported clinical and field cases. To verify the role of glutamine synthetase (GS) in the pathogenesis of S. xylosus, we evaluated the virulence level of the wild-type strain and its glnA mutant strain in biofilm assays in vitro and murine infection model in vivo. From the results, it was observed that the glnA mutant strain was attenuated and could reduce tissue damage. 1-Hydroxyanthraquinone (1-HAQ) is a kind of anthraquinones, it exhibited a significant inhibitory effect on the growth of S. xylosus and biofilm formation in vitro and provided anti-inflammatory effects in vivo. In addition, the rate at which it inhibits the biofilm, inflammatory factors, and CFU of wild-type strains were significantly higher than that of the mutant strains, indicating that 1-hAQ might have pharmacological effects against S. xylosus through the regulation of GS protein. The effect of 1-hAQ on GS was further confirmed by the down-regulation of glnA expression, reduced GS activity, Gln content and the results of molecular docking. Taken together, these findings suggest that 1-hAQ facilitated a significant attenuation of S. xylosus pathogenicity by regulating the GS protein: a vital virulence factor. Therefore, it can be inferred that 1-hAQ may serve as a potential source of organic compound for the development of novel alternative drugs in mitigating the menace of bovine mastitis.


Assuntos
Antraquinonas/farmacologia , Antibacterianos/farmacologia , Glutamato-Amônia Ligase/metabolismo , Staphylococcus/enzimologia , Staphylococcus/patogenicidade , Fatores de Virulência/metabolismo , Animais , Antraquinonas/química , Antraquinonas/uso terapêutico , Antibacterianos/uso terapêutico , Biofilmes/efeitos dos fármacos , Contagem de Colônia Microbiana , Modelos Animais de Doenças , Feminino , Mastite/tratamento farmacológico , Mastite/microbiologia , Mastite/patologia , Camundongos Endogâmicos BALB C , Testes de Sensibilidade Microbiana , Simulação de Acoplamento Molecular , Staphylococcus/efeitos dos fármacos , Staphylococcus/crescimento & desenvolvimento
7.
J Dairy Sci ; 103(3): 2693-2700, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31980229

RESUMO

The identification of milk microbial communities in ruminants is relevant for understanding the association between milk microbiota and health status. The most common approach for studying the microbiota is amplifying and sequencing specific hypervariable regions of the 16S rRNA gene using massive sequencing techniques. However, the taxonomic resolution is limited to family and, in some cases, genus level. We aimed to improve taxonomic classification of the water buffalo milk microbiota by amplifying and sequencing the full-length 16S rRNA gene (1,500 bp) using Nanopore sequencing (single-molecule sequencing). When comparing with short-read results, we improved the taxonomic classification, reaching species level. We identified the main microbial agents of subclinical mastitis at the species level that were in accordance with the microbiological culture results. These results confirm the potential of single-molecule sequencing for in-depth analysis of microbial populations in dairy animals.


Assuntos
Búfalos/microbiologia , Mastite/veterinária , Microbiota/genética , Leite/microbiologia , Sequenciamento por Nanoporos/veterinária , Animais , Feminino , Mastite/microbiologia , RNA Ribossômico 16S/genética
8.
J Anim Breed Genet ; 137(1): 73-83, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31602717

RESUMO

This work aimed to use 16S ribosomal RNA sequencing with the Illumina MiSeq platform to describe the milk microbiota from 50 healthy Assaf ewes. The global observed microbial community for clinically healthy milk samples analysed was complex and showed a vast diversity. The core microbiota of the sheep milk includes five genera: Staphylococcus, Lactobacillus, Corynebacterium, Streptococcus and Escherichia/Shigella. Although there are some differences, some of these genera are common with the microbiota core pattern of milk from other species, especially with dairy cows. The microbial composition of the studied samples, based on the definition of amplicon sequence variants, was analysed through a correlation network. A preliminary analysis by grouping the milk samples based on their somatic cell count (SCC), which is considered an indicator of subclinical mastitis (SM), showed certain differences for the core of the samples identified as SM. The differences in the microbiota diversity pattern among samples might also suggest that subclinical mastitis would be associated with the significant increase in some genera that are inhabitants of the mammary gland and a remarkable concomitant reduction in the microbial diversity. Additionally, we have also presented here a preliminary analysis to assess the impact of the sheep milk microbiome on SCC, as an indicator of subclinical mastitis. The results here reported provide a first characterization of the sheep milk microbiota and settle the basis for future studies in this field.


Assuntos
Microbiota/genética , Leite/microbiologia , RNA Ribossômico 16S/genética , Análise de Sequência de RNA , Ovinos/microbiologia , Animais , Contagem de Células , Classificação , Feminino , Mastite/microbiologia , Fenótipo , Ovinos/metabolismo
9.
J Dairy Sci ; 103(1): 890-897, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31733855

RESUMO

Mastitis in ruminants is an important disease with major effects on both the economy and animal welfare. It is caused by major pathogens such as Staphylococcus aureus and minor pathogens such as coagulase-negative staphylococci. The objective of this study was to identify and characterize staphylococci as a cause of sheep mastitis in Algeria. In this study, 123 milk samples were collected directly from the udder of sheep suffering from clinical mastitis in 2 provinces in Algeria. Recovered isolates were identified using MALDI-TOF mass spectrometry. Virulence-associated and antimicrobial resistance genes as well as clonal complexes (CC) of S. aureus were determined using microarray-based analysis. A total of 45 staphylococci isolates were cultivated from sheep milk samples, and 28 S. aureus were identified as methicillin susceptible (62.2%). Seventeen other Staphylococcus isolates of different species were identified using MALDI-TOF mass spectrometry. Subsequent microarray analysis typed the methicillin-susceptible S. aureus to 6 CC: CC8-MSSA, CC97-MSSA, CC130/521-MSSA, CC479-MSSA, CC522-MSSA, and CC705-MSSA. The accessory gene regulator agrIII and the ruminant leukocidin genes lukF-P83 and lukM were found in all isolates of CC130/521, CC479, CC522, and CC705. The toxic shock syndrome toxin gene tst1 was detected exclusively in CC130/521. Additionally, virulence-associated genes (sea, sed, sak, hld, hlgA, edinB, and others) were detected. The presence of antibiotic resistance genes [blaZ, erm(B), and tet(K)] was detected in small numbers of staphylococci. Staphylococci possessing these genes are considered potential hazards for farm animals, farmers, and consumers. Data concerning the prevalence and diversity of staphylococci causing mastitis in sheep from Algeria are lacking. Presented results on different aspects about staphylococci in Algerian sheep populations should at least partially close that gap. However, further extensive studies covering more geographical regions are needed to assess the epidemiological risk.


Assuntos
Mastite/veterinária , Doenças dos Ovinos/microbiologia , Infecções Estafilocócicas/veterinária , Staphylococcus/isolamento & purificação , Argélia , Animais , Toxinas Bacterianas/genética , Enterotoxinas/genética , Feminino , Leucocidinas/genética , Mastite/microbiologia , Meticilina/farmacologia , Leite/microbiologia , Ovinos , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/veterinária , Infecções Estafilocócicas/microbiologia , Staphylococcus/genética , Staphylococcus/patogenicidade , Superantígenos/genética , Virulência/genética
10.
Prev Vet Med ; 174: 104815, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31731036

RESUMO

Numerous factors affect milk somatic cell score (SCS) in dairy goats including physiologic factors such as stage of lactation, and pathological factors such as bacterial intramammary infection (IMI), e.g. staphylococcal IMI. The association between IMI and udder inflammation and especially differences in the inflammatory response between the different staphylococcal species are yet to be identified. The objectives of this study were to use data from a recently published longitudinal study that characterized IMI status over time to evaluate the species-specific effect of staphylococcal IMI on udder half-level milk SCS, and to evaluate the effect of time since IMI diagnosis on this relationship. Half-level milk samples for culture and determination of SCS were collected from 171 goats starting within 10 days in milk (DIM), and then at various intervals until ≥ 120 DIM. A mixed linear model was used to assess the effect of species-specific staphylococcal IMI on SCS, accounting for confounding by lentivirus infection, parity, DIM, and the contralateral half IMI status, and for repeated measurements within udder half and for clustering of halves within goat. A second model was used to evaluate the effect of time since first IMI diagnosis and its interaction with staphylococcal IMI status on the relationship between staphylococcal IMI status and SCS. Lentivirus seropositivity and contralateral half IMI status were associated with SCS. Staphylococcal IMI status was associated with SCS, with differences between species. Staphyloccocus caprae, Staphylococcus epidermidis, Staphylococcus simulans, and Staphylococcus xylosus were associated with higher SCS than a number of other species. Time since first diagnosis did not modify the relationship between IMI status and SCS.


Assuntos
Glândulas Mamárias Animais/microbiologia , Mastite/veterinária , Leite/microbiologia , Infecções Estafilocócicas/veterinária , Staphylococcus/isolamento & purificação , Animais , Contagem de Células/veterinária , Feminino , Doenças das Cabras/microbiologia , Doenças das Cabras/fisiopatologia , Cabras , Mastite/microbiologia , Mastite/fisiopatologia , Especificidade da Espécie , Infecções Estafilocócicas/microbiologia , Infecções Estafilocócicas/fisiopatologia
11.
Vet Microbiol ; 239: 108480, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31767091

RESUMO

Although coagulase-negative staphylococci are the primary aetiological agents of subclinical mastitis in ewes, there is little information regarding vaccination against that infection. The objective of this study was to evaluate the efficacy of a vaccine against staphylococcal mastitis in ewes under experimental conditions. The antigen in the vaccine is based on a bacterin of Staphylococcus aureus strain, expressing the exopolysaccharide poly-N-acetylglucosamine (PNAG), which is involved in biofilm formation by these bacteria. Ewes in groups A (n = 17) or B (n = 6) were given an initial vaccination 5 weeks before expected lambing, followed by a repeat administration 21 days later. Ewes in groups C (n = 8) or D (n = 6) were unvaccinated controls. Ewes in group A (n = 17) or C (n = 8) were challenged with a biofilm-forming S. chromogenes; animals in subgroups A1 or C1 were challenged on the 10th and those in A2 or C2 on the 50th day after lambing. Ewes in groups B or D were uninoculated controls. Clinical examinations of ewes, ultrasonographic examinations of udder, milk yield measurements, blood sampling for detection of anti-PNAG specific antibodies and milk sample collection for bacteriological and cytological examinations were performed up to 52nd day post-challenge. Finally, biopsies were performed for mammary tissue collection for histopathological examination. Among group A ewes, 29% developed systemic signs and 59% signs in the inoculated gland; the respective figures for group C were 50% and 100% (P =  0.040 for mammary signs). The median total clinical score was 2.0 for A and 5.5 for C ewes (P =  0.025). For A, but not for C, clinical scores decreased progressively during the study (P =  0.018 and P =  0.47, respectively). The duration of mastitis was shorter in A (4 days) than in C (17.5 days) ewes (P =  0.022). Bacterial counts were lower in milk samples from A than from C ewes, for samples collected from the inoculated and the uninoculated (P <  0.01) mammary glands of these ewes. Somatic cell counts in samples from inoculated and uninoculated mammary glands of A ewes were higher than in samples of C ewes (P <  0.02). There were differences for gray-scale evaluations during ultrasonographic examination and for milk yield measurements between groups (P <  0.01). Median bacterial counts in tissue samples from A ewes (0 cfu g-1) were lower than in ones from C (6.5 cfu g-1) ewes (P =  0.041). The median score for histopathological findings in tissue samples from inoculated glands of A was lower than that for C ewes: 1 versus 2 (P =  0.014). It is concluded that mastitis was less severe in vaccinated animals, as indicated by a wide array of measures.


Assuntos
Vacinas Bacterianas/normas , Biofilmes , Mastite/veterinária , Doenças dos Ovinos/prevenção & controle , Animais , Feminino , Glândulas Mamárias Animais/microbiologia , Mastite/microbiologia , Mastite/patologia , Mastite/prevenção & controle , Leite/citologia , Leite/microbiologia , Ovinos , Doenças dos Ovinos/microbiologia
12.
Arch Gynecol Obstet ; 300(6): 1637-1644, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31631250

RESUMO

PURPOSE: Although puerperal mastitis is a common disease, published data are poor. Increasing rates of community-acquired MRSA (CA-MRSA) cases are reported in the USA. However, information about common pathogens and CA-MRSA in Germany is still insufficient. The aim of this study was to investigate the most common pathogens of puerperal mastitis in the last decade, its therapy, resistance rate and the effectiveness of the current treatment strategies. METHODS: The pathogens, the respective antibiograms and the treatment strategies of patients treated for puerperal mastits at the University Clinic Magdeburg (Germany) between 2006 and 2016 were retrospectively reviewed. Statistical analysis was performed using SPSS Version 21. RESULTS: In our series, 59 cases with puerperal mastitis were reviewed, 26 (44.1%) of these developed a breast abscess. In 37 of 59 (67.3%) cases the symptoms occurred in the first 8 weeks postpartum. The most common pathogens were Staphylococcus aureus (64.9%) and Coagulase-negative Staphylococcus (13.5%). Methicillin-resistant Staphylococcus aureus (MRSA) was found in one case. Of the 19 cases with Staphylococcus aureus, 17 were resistant to Penicillin. Conservative management was mainly performed with Flucloxacillin (60%), which was successful in most cases. Sixteen of 26 (61.5%) women with abscesses underwent ultrasound (US)-guided needle aspiration. CONCLUSION: In this cohort, MRSA was not a main pathogen responsible for breast abscesses. Conservative treatment strategies remained constant during the observed period and Flucloxacillin was the most frequent antibiotic used. The analysis of the courses of diseases leads to the conclusion that surgical incision is progressively replaced by US-guided needle aspiration.


Assuntos
Mastite/tratamento farmacológico , Transtornos Puerperais/tratamento farmacológico , Abscesso/tratamento farmacológico , Abscesso/microbiologia , Adulto , Antibacterianos/uso terapêutico , Feminino , Humanos , Mastite/microbiologia , Transtornos Puerperais/microbiologia , Estudos Retrospectivos , Staphylococcus aureus/isolamento & purificação
13.
Transgenic Res ; 28(5-6): 573-587, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31599375

RESUMO

EchAMP, the tenth most abundant transcript expressed in the mammary gland of echidna, has in vitro broad-spectrum antibacterial effects. However, the effects of EchAMP on mastitis, a condition where inflammation is triggered following mammary gland infection, has not been investigated. To investigate the impact of EchAMP against mastitis, EchAMP transgenic mice were generated. In antibacterial assays, the whey fractions of milk from transgenic mice significantly reduced growth of Staphylococcus aureus, Bacillus subtilis, Escherichia coli and Pseudomonas aeruginosa compared with whey fractions from wildtype mice. Furthermore, a mastitis model created by infecting mammary gland with these four bacterial strains displayed a significant reduction in bacterial load in transgenic mice injected with S. aureus and B. subtilis. On further confirmation, histomorphologic analysis showed absence of necrosis and cell infiltration in the mammary glands of transgenic mice. To understand the role of EchAMP against inflammation, we employed an LPS-injected mastitis mouse model. LPS is known to induce phopshorylation of NF-κB and MAPK pathways, which in turn activate downstream proinflammatory signaling mediators, to promote inflammation. In LPS-treated EchAMP transgenic mice, phosphorylation levels of NF-κB, p38 and ERK1/2 were significantly downregulated. Furthermore, in mammary gland of transgenic mice, there was a significant downregulation of mRNA levels of proinflammatory cytokines, namely TNF-α, IL-6 and IL-1ß. Taken together, these data suggest that EchAMP has an antiinflammatory response and is effective against S. aureus and B. subtilis. We suggest that EchAMP may be a potential prophylactic protein against mastitis in dairy animals by expressing this gene in their mammary gland.


Assuntos
Peptídeos Catiônicos Antimicrobianos/genética , Inflamação/genética , Mastite/genética , Infecções Estafilocócicas/genética , Animais , Equidna/genética , Feminino , Humanos , Inflamação/induzido quimicamente , Inflamação/microbiologia , Inflamação/prevenção & controle , Interleucina-1beta/genética , Interleucina-6/genética , Lipopolissacarídeos/toxicidade , Sistema de Sinalização das MAP Quinases/genética , Glândulas Mamárias Animais/metabolismo , Mastite/induzido quimicamente , Mastite/microbiologia , Mastite/prevenção & controle , Camundongos , Camundongos Transgênicos/genética , NF-kappa B/genética , Infecções Estafilocócicas/microbiologia , Infecções Estafilocócicas/prevenção & controle , Staphylococcus aureus/genética , Staphylococcus aureus/patogenicidade , Fator de Necrose Tumoral alfa/genética
14.
Food Funct ; 10(10): 6543-6555, 2019 Oct 16.
Artigo em Inglês | MEDLINE | ID: mdl-31545328

RESUMO

Mastitis, a major disease affecting dairy cows, is most commonly caused by Staphylococcus aureus (S. aureus). Selenium (Se) can activate pivotal proteins in immune responses and regulate the immune system, and microRNA-155 (miR-155) is a key transcriptional regulator for inflammation-related diseases. We constructed the model of mouse mastitis in vivo and primary mouse mammary epithelial cells (MMECs) in vitro, which were induced by S. aureus. Se content of the mammary was estimated using an atomic fluorescence spectrophotometer. Histopathological analysis was performed via hematoxylin and eosin (H&E) staining. The mmu-miR-155-5p mimic was transfected in MMECs, and viability was determined through the MTT assay. Transfected efficiency was evaluated by qPCR and fluorescence staining. Cytokines including TNF-α, IL-1ß, IL-10 and TLRs were detected with qPCR. In addition, western blotting was used to evaluate the expression of the NF-κB and MAPKs signaling pathways. The results demonstrated that a Se-supplemented diet improved the content of Se in mammary tissues. Histopathological studies indicated that the mammary glands were protected in the Se-supplemented group after S. aureus infection. Se-supplementation suppressed the production of MPO, mmu-miR-155, TNF-α, IL-1ß, and TLR2 and significantly inhibited the phosphorylation of NF-κB and MAPKs in vivo and in vitro. All the data indicated that mmu-miR-155 played a pro-inflammatory role in our study, and Se-supplementation could suppress the expression of mmu-miR-155 to inhibit inflammation in S. aureus-induced mastitis in mice.


Assuntos
Doenças dos Bovinos/tratamento farmacológico , Mastite/tratamento farmacológico , MicroRNAs/genética , Selênio/administração & dosagem , Infecções Estafilocócicas/veterinária , Animais , Bovinos , Doenças dos Bovinos/genética , Doenças dos Bovinos/imunologia , Doenças dos Bovinos/microbiologia , Citocinas/genética , Citocinas/imunologia , Feminino , Regulação da Expressão Gênica , Mastite/genética , Mastite/imunologia , Mastite/microbiologia , Camundongos , MicroRNAs/imunologia , NF-kappa B/genética , NF-kappa B/imunologia , Infecções Estafilocócicas/genética , Infecções Estafilocócicas/imunologia , Infecções Estafilocócicas/microbiologia , Staphylococcus aureus/fisiologia , Receptor 2 Toll-Like/genética , Receptor 2 Toll-Like/imunologia
15.
Sci Rep ; 9(1): 13536, 2019 09 19.
Artigo em Inglês | MEDLINE | ID: mdl-31537825

RESUMO

Milk microbiomes significantly influence the pathophysiology of bovine mastitis. To assess the association between microbiome diversity and bovine mastitis, we compared the microbiome of clinical mastitis (CM, n = 14) and healthy (H, n = 7) milk samples through deep whole metagenome sequencing (WMS). A total of 483.38 million reads generated from both metagenomes were analyzed through PathoScope (PS) and MG-RAST (MR), and mapped to 380 bacterial, 56 archaeal, and 39 viral genomes. We observed distinct shifts and differences in abundance between the microbiome of CM and H milk in phyla Proteobacteria, Bacteroidetes, Firmicutes and Actinobacteria with an inclusion of 68.04% previously unreported and/or opportunistic strains in CM milk. PS identified 363 and 146 bacterial strains in CM and H milk samples respectively, and MR detected 356 and 251 bacterial genera respectively. Of the identified taxa, 29.51% of strains and 63.80% of genera were shared between both metagenomes. Additionally, 14 archaeal and 14 viral genera were found to be solely associated with CM. Functional annotation of metagenomic sequences identified several metabolic pathways related to bacterial colonization, proliferation, chemotaxis and invasion, immune-diseases, oxidative stress, regulation and cell signaling, phage and prophases, antibiotic and heavy metal resistance that might be associated with CM. Our WMS study provides conclusive data on milk microbiome diversity associated with bovine CM and its role in udder health.


Assuntos
Mastite Bovina/genética , Mastite Bovina/microbiologia , Microbiota/genética , Animais , Archaea/genética , Bactérias/genética , Biodiversidade , Bovinos/microbiologia , Feminino , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Glândulas Mamárias Animais/microbiologia , Mastite/genética , Mastite/microbiologia , Metagenoma , Metagenômica/métodos , Microbiota/fisiologia , Leite/microbiologia , Filogenia , RNA Ribossômico 16S/genética
16.
Microb Pathog ; 137: 103755, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31542423

RESUMO

Non-steroidal anti-inflammatory drugs (NSAIDs) may exhibit antibacterial activity and have synergistic effects with antibiotics. One way to re-sensitize MRSA to resistant antibiotics is by combining with approved non-antibiotics. The study was intended to explore susceptibility of MRSA to various antibiotics and non-antibiotics (NSAIDs) by micro dilution broth method. MRSA isolates were confirmed by PCR (mecA gene) and in-vitro antibiotic susceptibility was determined by disc diffusion tests. Combinations of NSAIDs with resistant antibiotics were also evaluated in in-vivo trial in mice. In-vitro antibiotic susceptibility profile presented 100% resistance to Cefoxitin, 37.5% to Gentamicin and Amikacin, 25% to Oxytetracycline, 12.5% to Tylosin, Fusidic acid and Vancomycin, while Ciprofloxacin, Levofloxacin, Moxifloxacin, Trimethoprim + Sulfamethoxazole and Linezolid were found sensitive. Synergistic effect was observed when resistant antibiotics (Oxytetracycline/Gentamicin) were combined with sensitive antibiotics (Ciprofloxacin/Linezolid). Combination of Non-antibiotics (NSAIDs) with resistant antibiotics revealed that Meloxicam showed partial synergism with both Oxytetracycline and Gentamicin, while Flunixin Meglumine presented synergistic effect with Oxytetracycline and partial synergism with Gentamicin. Diclofenac Sodium revealed additive effect with Oxytetracycline while in case of Gentamicin indifferent effect was observed. In-vivo results showed that combinations of Oxytetracycline with Meloxicam/Flunixin Meglumine and Gentamicin with Meloxicam/Flunixin Meglumine were effective. The study concluded that the resistance against milk borne zoonotic MRSA infections can be successfully addressed by combining resistant antibiotics with NSAIDs. Flunixin Meglumine can be used in combination with oxytetracycline against MRSA infection.


Assuntos
Antibacterianos/farmacologia , Anti-Inflamatórios não Esteroides/farmacologia , Gentamicinas/farmacologia , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Oxitetraciclina/farmacologia , Infecções Estafilocócicas/veterinária , Amicacina , Animais , Proteínas de Bactérias/genética , Combinação de Medicamentos , Sinergismo Farmacológico , Feminino , Cabras , Mastite/microbiologia , Mastite/veterinária , Staphylococcus aureus Resistente à Meticilina/genética , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Camundongos , Camundongos Endogâmicos BALB C , Testes de Sensibilidade Microbiana , Leite/microbiologia , Paquistão , Proteínas de Ligação às Penicilinas/genética , Prevalência , Infecções Estafilocócicas/tratamento farmacológico
17.
J Agric Food Chem ; 67(39): 10863-10870, 2019 Oct 02.
Artigo em Inglês | MEDLINE | ID: mdl-31507180

RESUMO

Mastitis, inflammation of the mammary gland, occurs in both humans and animals. Staphylococcus aureus is the most common infectious bacterial pathogen associated with mastitis. We investigated the effects of allicin on S. aureus-induced mastitis in mice. Pathological histology revealed that allicin inhibited S. aureus-induced pathological damage and myeloperoxidase activity in mammary tissues. Enzyme-linked immunosorbent assays demonstrated that allicin reduced the production of IL-1ß and TNF-α as well as inhibited the NF-κB and mitogen-activated protein kinase pathway by reducing phosphorylation of p65, IκBα, p38, JNK, and ERK. Western blotting revealed that allicin reduced TLR2 and TLR6 expression in mammary tissues and cells but not in HEK293 cells. The lipid raft content was reduced by allicin, which inhibited signaling downstream of TLR2 and TLR6. Liver X receptor α (LXRα) luciferase reporter assays and LXRα interference experiments showed that allicin improved the LXRα activity and adenosine 5'-triphosphate-binding cassette G and A1 (ABCG and ABCA1) expression, thereby reducing the cholesterol level, lipid raft formation, and downstream TLR2 and TLR6 pathway activity. These results demonstrated that allicin exerted anti-inflammatory effects against S. aureus mastitis by improving the LXRα activity and reducing lipid raft formation.


Assuntos
Mastite/tratamento farmacológico , Microdomínios da Membrana/efeitos dos fármacos , Infecções Estafilocócicas/tratamento farmacológico , Staphylococcus aureus/efeitos dos fármacos , Ácidos Sulfínicos/administração & dosagem , Animais , Feminino , Humanos , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Receptores X do Fígado/genética , Receptores X do Fígado/metabolismo , Mastite/genética , Mastite/metabolismo , Mastite/microbiologia , Microdomínios da Membrana/química , Microdomínios da Membrana/genética , Microdomínios da Membrana/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , NF-kappa B/genética , NF-kappa B/metabolismo , Infecções Estafilocócicas/genética , Infecções Estafilocócicas/metabolismo , Infecções Estafilocócicas/microbiologia , Staphylococcus aureus/fisiologia , Receptor 2 Toll-Like/genética , Receptor 2 Toll-Like/metabolismo
18.
Microb Pathog ; 137: 103732, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31505264

RESUMO

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.


Assuntos
Cefalosporinas/farmacologia , Cefalosporinas/uso terapêutico , Mastite/tratamento farmacológico , Mastite/microbiologia , Leite/microbiologia , Estresse Oxidativo , Animais , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Corynebacterium/isolamento & purificação , Feminino , Qualidade dos Alimentos , Glândulas Mamárias Animais/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Ovinos , Doenças dos Ovinos/tratamento farmacológico , Doenças dos Ovinos/microbiologia , Infecções Estafilocócicas/tratamento farmacológico , Infecções Estafilocócicas/microbiologia , Infecções Estafilocócicas/veterinária , Staphylococcus epidermidis/isolamento & purificação , Staphylococcus hyicus/isolamento & purificação , Staphylococcus intermedius/isolamento & purificação , Infecções Estreptocócicas/tratamento farmacológico , Infecções Estreptocócicas/microbiologia , Infecções Estreptocócicas/veterinária , Resultado do Tratamento
19.
BMC Genomics ; 20(1): 623, 2019 Jul 31.
Artigo em Inglês | MEDLINE | ID: mdl-31366318

RESUMO

BACKGROUND: Epigenetic changes such as cytosine (CpG) DNA methylations regulate gene expression patterns in response to environmental cues including infections. Microbial infections induce DNA methylations that play a potential role in modulating host-immune response. In the present study, we sought to determine DNA methylation changes induced by the mastitis causing Escherichia coli (E. coli) in porcine mammary epithelial cells (PMEC). Two time points (3 h and 24 h) were selected based on specific transcriptomic changes during the early and late immune responses, respectively. RESULTS: DNA methylation analysis revealed 561 and 898 significant (P < 0.01) differentially methylated CpG sites at 3 h and 24 h after E. coli challenge in PMEC respectively. These CpG sites mapped to genes that have functional roles in innate and adaptive immune responses. Significantly, hypomethylated CpG sites were found in the promoter regions of immune response genes such as SDF4, SRXN1, CSF1 and CXCL14. The quantitative transcript estimation indicated higher expression associated with the DNA CpG methylation observed in these immune response genes. Further, E. coli challenge significantly reduced the expression levels of DNMT3a, a subtype of de novo DNA methylation enzyme, in PMEC indicating the probable reason for the hypomethylation observed in the immune response genes. CONCLUSIONS: Our study revealed E. coli infection induced DNA methylation loci in the porcine genome. The differentially methylated CpGs were identified in the regulatory regions of genes that play important role in immune response. These results will help to understand epigenetic mechanisms for immune regulation during coliform mastitis in pigs.


Assuntos
Metilação de DNA/imunologia , Escherichia coli/fisiologia , Loci Gênicos/genética , Glândulas Mamárias Animais/imunologia , Glândulas Mamárias Animais/metabolismo , Animais , Epigenômica , Feminino , Perfilação da Expressão Gênica , Interações Hospedeiro-Patógeno/genética , Glândulas Mamárias Animais/citologia , Glândulas Mamárias Animais/microbiologia , Mastite/genética , Mastite/imunologia , Mastite/microbiologia , Suínos
20.
Int J Infect Dis ; 87: 135-142, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31442629

RESUMO

BACKGROUND: Breast tuberculosis (TB) is a rare disease, still present in endemic areas. The significance of breast TB is due to its rare occurrence and its resemblance to malignant breast lesions. The objective of this study was to study various clinical presentations and imaging features of different forms of tuberculous mastitis. METHODS: A retrospective study was conducted over a 10-year period. Thirty-two consecutive patients diagnosed with tuberculous mastitis were included. RESULTS: There was only one male patient in this series. The patients ranged in age from 25 to 60 years (mean age was 33.69 years). A solitary breast mass was the most common clinical presentation, which was associated with inflammatory findings in the majority of cases. The most common mammographic finding was diffusely increased density and the most common pathological finding was solitary mass. The most common appearance on ultrasound was of multiple intercommunicating breast masses with axillary lymphadenopathy. CONCLUSIONS: Breast TB should be considered in the differential diagnosis in the presence of a painful breast mass, discharging sinuses, mastitis, or a breast abscess that does not respond to conventional medical treatment. A multidisciplinary approach is required to aid timely diagnosis and to provide appropriate management and treatment in order to avoid complications.


Assuntos
Mastite/microbiologia , Tuberculose/microbiologia , Adulto , Mama/microbiologia , Mama/patologia , Diagnóstico Diferencial , Feminino , Humanos , Masculino , Mastite/diagnóstico , Mastite/patologia , Pessoa de Meia-Idade , Estudos Retrospectivos , Tuberculose/diagnóstico , Tuberculose/patologia
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA