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1.
Vet Clin North Am Food Anim Pract ; 37(1): 149-174, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33541696

RESUMO

Staphylococcus aureus is the most important cause of clinical mastitis in goats, and non-aureus staphylococci is the most common isolate from subclinical mastitis. Environmental streptococci are a severe problem. Somatic cell counts and California mastitis test are a screening test for mastitis and an indicator of poor udder health, but values should be interpreted differently than with dairy cattle. Somatic cell scores likely are a more useful way of viewing data. High bacterial counts in milk are common; mastitis may be involved as a cause. Proper udder preparation, milking procedure, and postmilking management are key factors in mastitis control.


Assuntos
Criação de Animais Domésticos/métodos , Indústria de Laticínios/métodos , Doenças das Cabras/prevenção & controle , Glândulas Mamárias Animais/fisiologia , Mastite/veterinária , Animais , Bovinos , Feminino , Doenças das Cabras/diagnóstico , Doenças das Cabras/microbiologia , Cabras , Glândulas Mamárias Animais/microbiologia , Mastite/diagnóstico , Mastite/microbiologia , Mastite/prevenção & controle , Leite/microbiologia , Infecções Estafilocócicas/prevenção & controle , Infecções Estafilocócicas/veterinária
2.
An Acad Bras Cienc ; 92(4): e20200521, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33331447

RESUMO

Copaiba oil is a natural product used by Amazonian populations and recognized for its medicinal properties because it has significant antimicrobial activity for several pathogenic microorganisms. The present work aimed to evaluate and characterize the effect of natural oil produced by copaiba - Copaifera multijuga against multiresistant isolates of bubaline mastitis. The nitrocefin test was performed with isolates of Staphylococcus aureus from bubaline mastitis, which were 100% positive for beta-lactamase enzyme detection. Minimum Bactericidal Concentration (MBC) of 25% to 3.12% was obtained for Enterococcus faecalis and Escherichia coli and 50% and 25% for S. aureus, but Klebsiella pneumoniae and Bacillus subtilis were resistant. MBC with 12.5% and 6.25% oil were obtained for most multiresistant bubaline mastitis isolates from the states of Pernambuco, Ceará, Bahia and Alagoas. The results demonstrated the great potential of using copaiba natural oil in the treatment of buffalo mastitis.


Assuntos
Anti-Infecciosos , Fabaceae , Mastite , Brasil , Feminino , Humanos , Mastite/tratamento farmacológico , Mastite/veterinária , Testes de Sensibilidade Microbiana , Staphylococcus aureus
3.
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
4.
Arq. bras. med. vet. zootec. (Online) ; 72(3): 1047-1050, May-June, 2020. ilus
Artigo em Português | LILACS, VETINDEX | ID: biblio-1129749

RESUMO

The aim of the present study was to evaluate the antibacterial behavior of polypyrrole nanoparticles (PPy-NPs) in water against biofilm producer or not S. aureus isolated from cows and goats with mastitis. One hundred and thirty-eight isolates of S. aureus were initially evaluated for biofilm formation by spectrophotometry in microplates. In addition, the minimum inhibition concentration (MIC) and minimum bactericidal concentration (MBC) of PPy-NPs in water for planktonic S. aureus were determined. From the bovine samples analyzed, 5 (4.46%) S. aureus isolates showed a strong biofilm production, 17 (15.18%) moderate production, 36 (32.14%) with weak production and 54 (48.21%) did not produce biofilms. Strains from goats (26) showed no biofilm production in 18 (69.23%) strains and weak biofilm production in 8 (30.76%) strains. The MIC and MBC of S. aureus to PPy-NPs were found in the same concentration (125搭/mL) in all strains tested, regardless of biofilm production or not. This finding provides a new insight into the interaction between PPy-NPs and S. aureus, and will offer potential benefits for the control of mastitis.(AU)


Assuntos
Animais , Feminino , Bovinos , Pirróis/administração & dosagem , Staphylococcus aureus/efeitos dos fármacos , Cabras/microbiologia , Mastite/veterinária , Biofilmes , Antibacterianos/uso terapêutico
5.
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
6.
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
7.
J Dairy Sci ; 103(3): 2677-2684, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31954559

RESUMO

The aim of this study was to characterize the proteins present in milk whey from buffaloes with and without subclinical mastitis using a proteomic approach to identify differentially expressed proteins as potential biomarkers for this disease. Whey from Murrah buffaloes with subclinical mastitis was compared with whey from healthy animals using liquid chromatography-tandem mass spectrometry. The annotated protein databases for Bubalus bubalis and Bos taurus were used in the analysis, and the gene annotations from the buffalo and bovine reference assemblies were also used. After integrating gene annotations from both buffaloes and bovines, a total of 1,033 proteins were identified, of which 156 were differentially expressed. Eighteen biological processes were annotated with Gene Ontology. Cathelicidin-3 was identified as a potential biomarker for subclinical mastitis. These results are important to the characterization of mastitis in the buffalo mammary gland and may aid in the development of tools for early diagnosis.


Assuntos
Peptídeos Catiônicos Antimicrobianos/análise , Mastite/veterinária , Proteínas do Leite/análise , Proteômica , Soro do Leite/química , Animais , Biomarcadores/análise , Búfalos , Bovinos , Cromatografia Líquida/veterinária , Feminino , Mastite/metabolismo , Mastite Bovina/metabolismo , Espectrometria de Massas em Tandem/veterinária , Proteínas do Soro do Leite/análise
8.
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
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.
Res Vet Sci ; 128: 129-134, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31783263

RESUMO

A recently developed bovine cathelicidin (CATH) ELISA was evaluated in the dairy buffalo (Bubalus bubalis) by testing 618 quarter milk samples from a herd with subclinical mastitis cases. Somatic cell count (SCC) and bacteriological culture (BC) were carried out on the same samples for comparison. Out of 618 quarters, 258 (41.75%) were positive to CATH, 289 (46.76%) had SCC > 200,000 cells/mL, and 457 (73.95%) were positive to BC. The most prevalent microorganism was Staphylococcus aureus (SAU, 35.76% of all quarters), followed by non-aureus staphylococci (NAS, 22.17% of all quarters). Clinical mastitis quarters were only 7 (1.13%). CATH levels were significantly higher in clinical quarters and in high SCC, BC-positive quarters than in healthy, low SCC, BC-negative quarters. The highest median values were observed for SAU and the lowest for NAS. Differences among microorganism classes were generally more significant for SCC than for CATH. Test characteristics of the CATH ELISA, evaluated by considering as true positives all BC-positive quarters with SCC > 200,000 cells/mL (N = 242), and as true negatives all sterile quarters with SCC < 200,000 cells/mL (N = 44), were as follows: sensitivity 57.85%, specificity 84.09%, positive predictive value 95.24%, negative predictive value 26.62%, accuracy 61.89%. Therefore, the bovine CATH ELISA showed a fair sensitivity and a good specificity in detecting water buffalo mastitis.


Assuntos
Peptídeos Catiônicos Antimicrobianos/química , Búfalos , Ensaio de Imunoadsorção Enzimática/veterinária , Mastite/veterinária , Leite/química , Animais , Bovinos , Contagem de Células/veterinária , Testes Diagnósticos de Rotina , Ensaio de Imunoadsorção Enzimática/métodos , Ensaio de Imunoadsorção Enzimática/normas , Feminino , Mastite/diagnóstico , Leite/citologia , Leite/microbiologia , Sensibilidade e Especificidade , Infecções Estafilocócicas/diagnóstico , Infecções Estafilocócicas/veterinária , Staphylococcus aureus/crescimento & desenvolvimento
11.
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
12.
Vet Immunol Immunopathol ; 219: 109972, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31733501

RESUMO

This study aimed to determine if intrauterine-infused lipopolysaccharides (LPS) can be translocated to the mammary glands and induce an inflammatory response. Thirty-seven goats were divided into two experiments. Nineteen goats (control group, n = 9; LPS group, n = 10) were subjected to intravenous injection of LPS, and eighteen goats (control group, n = 8; LPS group, n = 10) were subjected to intrauterine infusion of LPS. Milk and blood samples were collected before and after the LPS challenge, to measure the blood leukocyte count (BLC), plasma LPS-binding protein (LBP), milk yield, milk somatic cell count (SCC), lactoferrin (LF), milk lactoperoxidase (LPO) activity, and pro- and anti-inflammatory cytokines in plasma and milk. Mammary gland tissues were collected from the parenchyma before and after the LPS challenge, for immunohistochemistry of LPS. In the intravenous injection experiment, the BLC (P < 0.001) and milk yield (P = 0.009) were lower, whereas the LF concentration (P < 0.001) and milk LPO activity (P < 0.001) were higher in the LPS group compared to that in the control group. LPS was detected in the mammary gland 3 and 24 h after intravenous injection of LPS. In the intrauterine infusion experiment, the mean concentrations of IL-1ß and IL-6 in milk were higher in the LPS group compared to that in the control group (P = 0.004 and P = 0.017, respectively), whereas there were no changes in milk yield or SCC. LPS was detected in the connective tissues and interepithelial spaces of the alveoli of the mammary glands 24 h after intrauterine infusion of LPS. We conclude that intrauterine-infused LPS can be translocated to the mammary glands from the uterus, however, the amount of translocated LPS might not be enough to induce symptoms of clinical or subclinical mastitis.


Assuntos
Lipopolissacarídeos/administração & dosagem , Glândulas Mamárias Animais/efeitos dos fármacos , Glândulas Mamárias Animais/imunologia , Mastite/veterinária , Leite/química , Útero/efeitos dos fármacos , Proteínas da Fase Aguda/análise , Administração Intravenosa , Animais , Proteínas de Transporte/análise , Feminino , Cabras/imunologia , Lactação/efeitos dos fármacos , Lactoferrina/análise , Lipopolissacarídeos/imunologia , Glicoproteínas de Membrana/análise , Leite/citologia
13.
Pesqui. vet. bras ; 39(12): 954-960, Dec. 2019. tab, graf, ilus
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1056928

RESUMO

The indiscriminate use of antibiotics in the treatment of caprine mastitis causes the appearance of resistant microorganisms, besides leaving residues in milk, putting at risk to human health. In this way, propolis is an alternative in the treatment of diseases because it has antimicrobial activity, mainly because of the presence of flavonoids in its composition. The aim of this study was to evaluate the antimicrobial potential of propolis to Staphylococcus spp. Isolated from cases of goat mastitis and qualify the crude ethanoic extract by high performance liquid chromatography (HPLC). In this study, the minimum bactericidal concentration values of propolis extracts in ethanol, ethyl acetate and hexane showed that the best concentrations capable of promoting the highest mortality of the isolates of Staphylococcus spp. from mastitis in goats, were 6250, 3125 and 1562.5µg/mL, respectively. By the microplate adherence test, it was found that 20.78% isolates were not able to form biofilm, 14.70% were classified as moderate and 64.70% were weak and none as a strong biofilm producer. Propolis in its different diluents was able to affect the formation of biofilm and showed a pronounced marked antimicrobial activity against Staphylococcus spp. strains and may be indicated for use in in vivo studies.(AU)


O uso indiscriminado de antibióticos no tratamento de mastite caprina leva ao desenvolvimento de micro-organismos resistentes que poderão estar presentes em alimentos, colocando em risco a saúde humana. Dessa forma, a própolis surge como uma alternativa para o tratamento de doenças por possuir uma ação antimicrobiana, principalmente pela presença de flavonoides em sua composição. O objetivo desse estudo foi avaliar o potencial antimicrobiano da própolis frente à Staphylococcus spp. isolados de casos de mastite caprina e qualificar o extrato etanoico bruto por cromatografia líquida de alta eficiência (CLAE-DAD). Neste estudo, os valores de concentração bactericida mínima (CBM) dos extratos de própolis em álcool etílico, acetato de etila e hexano nos isolados foram de 6250, 3125 e 1562,5µg/mL, respectivamente. Pelo teste de aderência à microplacas, observou-se que 20,78% dos microorganismos, não foram capazes de formar biofilme, 14,70% foram classificados como moderados, 64,70% em fracos e nenhum como forte produtor de biofilme. A própolis em seus diferentes diluentes foi capaz de afetar a formação de biofilme e apresentou significativa atividade antimicrobiana frente a cepas de Staphylococcus spp., podendo ser indicada para utilização em estudos "in vivo".(AU)


Assuntos
Animais , Feminino , Própole/uso terapêutico , Infecções Estafilocócicas/terapia , Infecções Estafilocócicas/veterinária , Staphylococcus/isolamento & purificação , Cabras/microbiologia , Apiterapia/veterinária , Mastite/terapia , Mastite/veterinária
14.
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
15.
PLoS One ; 14(11): e0214346, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31765378

RESUMO

Milk yield is the most important dairy sheep trait and constitutes the key genetic improvement goal via selective breeding. Mastitis is one of the most prevalent diseases, significantly impacting on animal welfare, milk yield and quality, while incurring substantial costs. Our objectives were to determine the feasibility of a concomitant genetic improvement programme for enhanced milk production and resistance to mastitis. Individual records for milk yield, and four mastitis-related traits (milk somatic cell count, California Mastitis Test score, total viable bacterial count in milk and clinical mastitis presence) were collected monthly throughout lactation for 609 ewes of the Chios breed. All ewes were genotyped with a mastitis specific custom-made 960 single nucleotide polymorphism (SNP) array. We performed targeted genomic association studies, (co)variance component estimation and pathway enrichment analysis, and characterised gene expression levels and the extent of allelic expression imbalance. Presence of heritable variation for milk yield was confirmed. There was no significant genetic correlation between milk yield and mastitis traits. Environmental factors appeared to favour both milk production and udder health. There were no overlapping of SNPs associated with mastitis resistance and milk yield in Chios sheep. Furthermore, four distinct Quantitative Trait Loci (QTLs) affecting milk yield were detected on chromosomes 2, 12, 16 and 19, in locations other than those previously identified to affect mastitis resistance. Five genes (DNAJA1, GHR, LYPLA1, NUP35 and OXCT1) located within the QTL regions were highly expressed in both the mammary gland and milk transcriptome, suggesting involvement in milk synthesis and production. Furthermore, the expression of two of these genes (NUP35 and OXCT1) was enriched in immune tissues implying a potentially pleiotropic effect or likely role in milk production during udder infection, which needs to be further elucidated in future studies. In conclusion, the absence of genetic antagonism between milk yield and mastitis resistance suggests that simultaneous genetic improvement of both traits be achievable.


Assuntos
Lactação/genética , Mastite/veterinária , Leite , Doenças dos Ovinos/genética , Carneiro Doméstico/genética , Animais , Indústria de Laticínios , Estudos de Viabilidade , Feminino , Genômica , Lactação/fisiologia , Mastite/genética , Análise de Sequência com Séries de Oligonucleotídeos , Polimorfismo de Nucleotídeo Único , Locos de Características Quantitativas , Seleção Artificial , Ovinos , Carneiro Doméstico/classificação , Carneiro Doméstico/fisiologia , Especificidade da Espécie
16.
PLoS One ; 14(9): e0217888, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31557171

RESUMO

The benefits and efficacy of control programs for herds infected with Mycobacterium avium subsp. paratuberculosis (MAP) have been investigated under various contexts. However, most previous research investigated paratuberculosis control programs in isolation, without modeling the potential association with other dairy diseases. This paper evaluated the benefits of MAP control programs when the herd is also affected by mastitis, a common disease causing the largest losses in dairy production. The effect of typically suggested MAP controls were estimated under the assumption that MAP infection increased the rate of clinical mastitis. We evaluated one hundred twenty three control strategies comprising various combinations of testing, culling, and hygiene, and found that the association of paratuberculosis with mastitis alters the ranking of specific MAP control programs, but only slightly alters the cost-benefit difference of particular MAP control components, as measured by the distribution of net present value of a representative U.S. dairy operation. In particular, although testing and culling for MAP resulted in a reduction in MAP incidence, that control led to lower net present value (NPV) per cow. When testing was used, ELISA was more economically beneficial than alternative testing regimes, especially if mastitis was explicitly modeled as more likely in MAP-infected animals, but ELISA testing was only significantly associated with higher NPV if mastitis was not included in the model at all. Additional hygiene was associated with a lower NPV per cow, although it lowered MAP prevalence. Overall, the addition of an increased risk of mastitis in MAP-infected animals did not change model recommendations as much as failing to consider.


Assuntos
Mastite/veterinária , Modelos Econométricos , Modelos Estatísticos , Mycobacterium avium subsp. paratuberculosis , Paratuberculose/epidemiologia , Paratuberculose/prevenção & controle , Algoritmos , Animais , Bovinos , Laticínios/economia , Feminino , Abastecimento de Alimentos , Leite , Medição de Risco , Fatores de Risco
17.
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
18.
J Dairy Sci ; 102(10): 9298-9311, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31421883

RESUMO

Mastitis is a highly prevalent condition that has a great impact on milk production and animal welfare, and often requires substantial management efforts. For this reason, it is generally considered an important threat to the dairy industry. Many microbial, host, and environmental factors can protect against, predispose to, or influence the development of mastitis. The objective of this work was to characterize the milk microbiota of Manchega ewes, and to compare samples from animals with and without a history of mastitis. We analyzed milk samples from 36 ewes belonging to 2 different farms (18 ewes from each farm) using culture-dependent and culture-independent techniques. We also analyzed several immune compounds to investigate associations of mastitis with 3 main variables: farm; history of mastitis or no mastitis; and parity number. Both culture-dependent and culture-independent techniques showed that ewe milk harbored a site-specific complex microbiota and microbiome. Staphylococcus epidermidis was the main species driving the difference between farm A (where it was the dominant species) and B (where it was not). In contrast, samples from farm B were characterized by the presence of a wide spectrum of other coagulase-negative staphylococci. Some of these species have already been associated with subclinical intramammary infections in ruminants. Of the 10 immune compounds assayed in this study, 3 were related to a history of mastitis [IL-8, IFN-γ, and IFN-gamma-induced protein 10 (IP-10)]. Increases in IL-8 concentrations in milk seemed to be a feature of subclinical mastitis in sheep, and in this study, this immune factor was detected only in samples from ewes with some episodes of mastitis and from the group with the highest somatic cell count. We also observed a positive correlation between the samples with the highest somatic cell count and IFN-γ and IP-10 levels. Our results suggest that these 3 compounds could be used as biomarkers for the negative selection of mastitis-prone animals, particularly when somatic cell count is very high.


Assuntos
Mastite/veterinária , Microbiota , Leite/microbiologia , Doenças dos Ovinos/microbiologia , Staphylococcus epidermidis , Animais , Contagem de Células/veterinária , Feminino , Leite/metabolismo , Paridade , Gravidez , Prevalência , Ovinos , Doenças dos Ovinos/imunologia , Staphylococcus/isolamento & purificação , Staphylococcus epidermidis/isolamento & purificação
19.
J Dairy Sci ; 102(10): 9328-9344, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31400892

RESUMO

Our objective was to evaluate the efficacy of a vaccine against staphylococcal mastitis in 5 dairy sheep farms, with 316 ewes in the vaccinated (V) group and 307 in the control (C) group studied throughout a lactation period. Two administrations of the vaccine were performed during the last stage of gestation of ewes. Starting 15 d after lambing and at monthly intervals thereafter, up to 9 milk samplings were performed for bacteriological and cytological examinations. Staphylococcal isolates recovered were examined for biofilm formation. Blood samples were collected for measurement of IgG poly-N-acetylglucosamine-specific antibodies. The most frequently isolated bacteria were staphylococci: 56.4 and 76.1%, respectively, of total isolates recovered from ewes of group V and C, respectively; staphylococci as causal agents of mastitis were isolated less frequently from V (5.3%) than in ewes in C (10.3%). Among mastitis-associated staphylococcal isolates recovered from V ewes, a smaller proportion was biofilm-forming than among ones from C: 53.2% versus 74.9% of isolates; biofilm-forming staphylococci as causal agents of mastitis were isolated less frequently from ewes in group V (2.3%) than in ewes in group C (6.0%). Anti-poly-N-acetylglucosamine-specific antibody values increased in V ewes and were higher than in C; a greater proportion of ewes with low antibody titers developed staphylococcal mastitis (41.4%) than of V ewes with high antibody titers (17.0%). Incidence risk of mastitis, staphylococcal mastitis, and biofilm-associated staphylococcal mastitis was smaller in V than in C: 36.7, 17.1, and 8.0% versus 44.3, 30.9, and 18.9%, respectively. The first case of staphylococcal mastitis occurred later in V than in C: third versus second sampling point. Overall, efficacy of the vaccine was 44.6% for staphylococcal mastitis, 57.7% for biofilm-associated staphylococcal mastitis, 33.1% for staphylococcal intramammary infection, and 51.5% for biofilm-associated staphylococcal intramammary infection. Nevertheless, vaccination should not be the only means for controlling mastitis; other udder health management measures should be included therein to improve control of the infection.


Assuntos
Vacinas Bacterianas/administração & dosagem , Biofilmes , Mastite/veterinária , Doenças dos Ovinos/prevenção & controle , Infecções Estafilocócicas/veterinária , Animais , Feminino , Incidência , Lactação , Glândulas Mamárias Animais/microbiologia , Mastite/prevenção & controle , Leite/microbiologia , Distribuição Aleatória , Ovinos , Doenças dos Ovinos/microbiologia , Infecções Estafilocócicas/prevenção & controle
20.
Comp Immunol Microbiol Infect Dis ; 66: 101339, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31437679

RESUMO

IL-1R8 is a member of Interleukin-1 receptor family acting as a negative regulator of inflammation reliant on ILRs and TLRs activation. IL-1R8 role has never been evaluated in acute bacterial mastitis. We first investigated IL-1R8 sequence conservation among different species and its pattern of expression in a wide panel of organs from healthy goats. Then, modulation of IL-1R8 during natural and experimental mammary infection was evaluated and compared in blood, milk and mammary tissues from healthy and Staphylococcus aureus infected goats. IL-1R8 has a highly conserved sequence among vertebrates. Goat IL-1R8 was ubiquitously expressed in epithelial and lymphoid tissues with highest levels in pancreas. IL-1R8 was down-regulated in epithelial mammary cells following S. aureus infection. Interestingly it was up-regulated in leukocytes infiltrating the infected mammary tissues suggesting that it could represent a target of S. aureus immune evasion.


Assuntos
Doenças das Cabras/imunologia , Imunidade Inata , Glândulas Mamárias Animais/microbiologia , Mastite/veterinária , Receptores de Interleucina-8/genética , Infecções Estafilocócicas/imunologia , Animais , Regulação para Baixo , Células Epiteliais/imunologia , Células Epiteliais/microbiologia , Feminino , Doenças das Cabras/microbiologia , Cabras/microbiologia , Inflamação , Glândulas Mamárias Animais/imunologia , Mastite/imunologia , Mastite/microbiologia , Receptores de Interleucina-8/sangue , Staphylococcus aureus/imunologia , Regulação para Cima
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