Speciation of coagulase negative staphylococci (CoNS) isolated from various clinical specimens in a tertiary care hospital of southern Rajasthan

Background: Coagulase negative staphylococci (CoNS) are a group of staph bacteria, which generally exist as normal flora of human skin and the oral flora found on mucous membranes. CoNS in immunocompromised pa?ents can cause variety of infec?ons like bacteraemia, central nervous system shunt infec?on, intravascular catheter-related infec?ons, endocardi?s, surgical site infec?ons, urinary tract infec?ons, foreign body infec?on, endophthalmi?s, peritoni?s and wound, bone and joint infec?ons as well as infec?ons in neonates. These infec?ons are difficult to treat because of the risk factors and the drug resistant nature of the organisms.Objec?ve:To find out species wise distribu?on of Coagulase nega?ve Staphylococci (CoNS) in various relevant clinical samples in our ins?tute. Materials and Method:A total of 500 CoNS strains were isolated from relevant clinical specimens. CoNS strains were isolated from variety of clinical specimens. Strains were iden?fied as CoNS on the basis of colony morphology, gram stain, catalase test and coagulase test. Conven?onal methods were used for species iden?fica?on a?er confirming isolates as CoNS. Results:The most common source of CoNS isola?on was blood (34%), followed by respiratory secre?ons (24.4%), urine (16.8%), pus (13.4%), swabs (5.4%), bodily fluid (1.4%), and others (4.6%). S.epidermidis was most frequently isolated (30.6%), followed by S.hemoly?cus (25.4%), S.hominis (12.6%), S.capi?s (12%), S.lugdunensis (11.4%) and S.cohnii (08%). Conclusion : As CoNS have become major cause of nosocomial infec?ons, there is a need for rapid iden?fica?on and specia?on of CoNS with their an?bio?c suscep?bility for be?er management of these cases and to prevent emergence of drug resistance.

Epidemiological Profile of Patients with Infective Endocarditis at three Tertiary Centers in Brazil from 2003 to 2017

Abstract Background Infective endocarditis (IE) is a disease with high morbimortality and an increasing incidence. With improved diagnosis and treatment, a number of epidemiological changes have been reported over time. Objectives We sought to describe the epidemiological profile, mortality predictors, and analysis of a possible microbiological transition in patients admitted to three tertiary centers in Brazil. Methods In this cross-sectional retrospective study, data from 211 patients with definite or probable IE were analyzed according to the modified Duke criteria between 2003 and 2017. The association between categorical variables was assessed using the chi-square or Fisher's exact test, and binary logistic models were built to investigate mortality. We considered p <0.05 statistically significant. Results The median age of the sample was 48 (33-59) years old, 70.6% were men, and the most prevalent pathogen was Staphylococcus spp. (19%). Mortality was 22.3%, with increasing age being the leading risk factor for death (p = 0.028). Regarding the location of the disease, native valves were the most affected site, with the aortic valve being more affected in men than women (p = 0.017). The mean number of cases of Staphylococcus spp. (τ = 0.293, p = 0.148) and Streptococcus spp. (τ = -0.078, p = 0.727) has remained stable over the years. Conclusion No trend towards reduced or increased mortality was evident between 2003 and 2017. Although Staphylococcus spp. were the most prevalent pathogen, the expected epidemiological transition could not be observed.

Infección por Staphylococcus lugdunensis: descripción de 44 casos / Staphylococcus lugdunensis infection: report of 44 cases

INTRODUCCIÓN: Staphylococcus lugdunensis, es un estafilococo coagulasa negativa (SCN) con características de virulencia y de sensibilidad antimicrobiana que lo hacen más parecido a Staphylococcus aureus que a otros SCN. OBJETIVOS: Conocer las características clínicomicrobiológicas de los aislados de S. lugdunensis identificados en nuestra institución. MATERIAL Y MÉTODOS: Se realizó un estudio retrospectivo de los aislados de S. lugdunensis entre los años 2017 y 2019 en el Servicio de Microbiología del Hospital Universitario San Jorge de Huesca (España). Se revisaron las historias clínicas correspondientes a los pacientes con aislamiento de S. lugdunensis, considerándose las siguientes variables: edad, sexo, tipo de muestra, servicio de procedencia y enfermedad de base. La identificación bacteriana se realizó con MALDI-TOF VITEK MS (BioMérieux, Francia). Así mismo, se estudió su patrón de susceptibilidad antimicrobiana in vitro mediante microdilución en placa. RESULTADOS: Se obtuvieron 44 aislados de S. lugdunensis: 12 procedían de heridas, 10 fueron abscesos, 8 úlceras, 7 orinas, 4 frotis cutáneos, 2 exudados óticos, y 1 exudado vaginal. En relación con la enfermedad de base destacaron cinco pacientes con procesos tumorales y diez con diabetes mellitus. En 17 pacientes existían antecedentes de cirugía o traumatismo reciente. La mayoría de las cepas fueron sensibles a los antimicrobianos estudiados. En 19 de ellas se observó producción de β-lactamasa, dos fueron resistentes a macrólidos y tres a clindamicina. Todas las cepas fueron sensibles a oxacilina, gentamicina y cotrimoxazol. CONCLUSIONES: Aunque S. lugdunensis mantiene una buena sensibilidad a la mayoría de los antimicrobianos, su tendencia a producir abscesos y que exprese factores de virulencia más parecido a S. aureus que a otros SCN, hace necesaria una correcta identificación en el laboratorio con el fin de que su incidencia no quede subestimada.

BACKGROUND: Staphylococcus lugdunensis is a coagulase-negative staphylococcus (CNS) with virulence and antibiotic sensitivity characteristics which makes it more similar to Staphylococcus aureus than other CNS. AIM: To know the microbiological and clinical characteristics of S. lugdunensis isolates identified from our health sector. METHODS: A retrospective study of S. lugdunensis isolates was carried out between 2017 and 2019 in the Microbiology Service of the San Jorge University Hospital in Huesca (Spain). The clinical records of patients with S. lugdunensis isolation were reviewed, considering the following factors: age, sex, sample type, service and underlying disease. Bacterial identification was performed using MALDI-TOF VITEK MS (BioMérieux, France). The pattern of antibiotic susceptibility was studied by means of plate microdilution. RESULTS: 44 isolates of S. lugdunensis were obtained: 12 corresponded to wounds, 10 were abscesses, 8 ulcers, 7 urine samples, 4 skin smears, 2 otic exudates, and 1 vaginal exudate. Regarding the underlying disease, five patients had a tumor processes and ten had diabetes mellitus. In 17 patients there was a history of recent surgery or trauma. Most of the strains were susceptible to the antibiotics studied. Production of beta-lactamase was observed in 19 of them, two were resistant to macrolides and three to clindamycin. None of the isolates were resistant to oxacillin, gentamicin or cotrimoxazole. CONCLUSIONS: Although S. lugdunensis maintains a good sensitivity to most antibiotics, its tendency to produce abscesses and that it expresses virulence factors more similar to S. aureus than to other CNS requires a correct identification in the laboratory so that its incidence is not underestimated.

Susceptibility to Vancomycin of Biofilm Producing Staphylococci Isolated from Tertiary Care Hospital of Nepal / Susceptibilidad a la vancomicina de estafilococos productores de biopelículas aislados del hospital de atención terciaria de Nepal

Background: Methicillin resistance and biofilm-producing Staphylococci are emerging as multidrug-resistant strains narrowing the efficacy of antimicrobial therapy. Although vancomycin is used as the drug of choice to treat such isolates, different studies worldwide have documented the emergence of strains that are intermediately susceptible or resistant to this antibiotic. Objective: The study aimed to determine the minimum inhibitory concentration of vancomycin to methicillin-resistant and biofilm-producing staphylococci isolated from different clinical specimens. Methods: 375 staphylococci isolated from different clinical specimens over one year were included in the study. Biofilm formation was determined by the Tissue culture plate method (TCP), and ica genes were identified by Polymerase Chain Reaction (PCR). Antibiotic susceptibility and methicillin resistance were done following Clinical and Laboratory Standards Institute (CLSI) guidelines. The minimum inhibitory concentration (MIC) of vancomycin in all isolates was determined by the agar dilution method. Results:Among 375 Staphylococci studied, 43% and 57% represented S. aureus and Coagulase-Negative Staphylococci (CNS), respectively. The rate of Methicillin-Resistant S. aureus (MRSA) and Methicillin-Resistant Coagulase Negative Staphylococci (MRCNS) were 81.4% and 66.8% respectively and determined by the disc diffusion method. The most potential antibiotics were tetracycline and chloramphenicol showing sensitivity to more than 90% isolates. The Minimum Inhibitory Concentration (MIC) value of oxacillin for staphylococci ranged from 0.125-32 µg/ml. Oxacillin agar diffusion method showed 51.6% and 79.9% isolates as MRSA and MRCNS, respectively, revealing a very high percentage of S. aureus and CNS isolates as methicillin-resistant. All isolates had susceptible vancomycin MICs that ranged from 0.125-2 µg/ml. Two S. aureus isolated from Central Venous Catheter (CVC) and catheter specimens were detected with intermediate susceptibility to vancomycin. Similarly, three CNS isolated from blood, CVC, and wound/pus (w/p) were intermediately susceptible to vancomycin. Strong biofilm formation was observed in 22.1% of clinical isolates, and the ica gene was detected among 22.9% of isolates. Only one S. aureus detected as a biofilm producer by the TCP method was found to have intermediate susceptibility to vancomycin. Conclusions: The increment in vancomycin MIC among methicillin-resistant and biofilm-producing staphylococci is alarming. Strict control measures to prevent methicillin-resistant isolates spread and routine surveillance for vancomycin-resistant isolates must be incorporated in hospitals to prevent antimicrobial treatment failure

Antecedentes: Los estafilococos resistentes a la meticilina y productores de biopelículas están surgiendo como cepas multirresistentes que reducen la eficacia del tratamiento antimicrobiano. Aunque la vancomicina se utiliza como fármaco de elección para tratar dichos aislados, diferentes estudios realizados en todo el mundo han documentado la aparición de cepas intermedias susceptibles o resistentes a este antibiótico. Objetivo: El estudio tenía como objetivo determinar la concentración mínima inhibitoria de la vancomicina para los estafilococos resistentes a la meticilina y productores de biofilm aislados de diferentes muestras clínicas. Métodos: Se incluyeron en el estudio 375 estafilococos aislados de diferentes muestras clínicas durante un año. La formación de biopelículas se determinó mediante el método de la placa de cultivo de tejidos (TCP), y los genes ica se identificaron mediante la reacción en cadena de la polimerasa (PCR). La susceptibilidad a los antibióticos y la resistencia a la meticilina se realizaron siguiendo las directrices del Clinical and Laboratory Standards Institute (CLSI). La concentración inhibitoria mínima (MIC) de vancomicina en todos los aislados se determinó por el método de dilución en agar. Resultados:Entre los 375 estafilococos estudiados, el 43% y el 57% representaban S. aureus y estafilococos coagulasa-negativos (ECN), respectivamente. La tasa de S. aureus resistente a la meticilina (SARM) y de estafilococos coagulasa negativos resistentes a la meticilina (ECNM) fue del 81,4% y el 66,8%, respectivamente, y se determinó por el método de difusión de discos. Los antibióticos más potenciales fueron la tetraciclina y el cloranfenicol, que mostraron una sensibilidad superior al 90% de los aislados. El valor de la concentración inhibitoria mínima (CIM) de la oxacilina para los estafilococos osciló entre 0,125-32 µg/ml. El método de difusión en agar de la oxacilina mostró que el 51,6% y el 79,9% de los aislados eran SARM y MRCNS, respectivamente, lo que revela que un porcentaje muy elevado de los aislados de S. aureus y CNS son resistentes a la meticilina. Todos los aislados tenían MIC de vancomicina susceptibles que oscilaban entre 0,125-2 µg/ml. Se detectaron dos S. aureus aislados de muestras de catéteres venosos centrales (CVC) y catéteres con una susceptibilidad intermedia a la vancomicina. Del mismo modo, tres S. aureus aislados de sangre, CVC y herida/pus (w/p) fueron intermedianamente susceptibles a la vancomicina. Se observó una fuerte formación de biopelículas en el 22,1% de los aislados clínicos, y se detectó el gen ica en el 22,9% de los aislados. Sólo un S. aureus detectado como productor de biopelículas por el método TCP resultó tener una susceptibilidad intermedia a la vancomicina. Conclusiones: El incremento de la MIC de vancomicina entre los estafilococos resistentes a la meticilina y productores de biofilm es alarmante. Para evitar el fracaso del tratamiento antimicrobiano, deben incorporarse en los hospitales medidas de control estrictas para prevenir la propagación de los aislados resistentes a la meticilina y una vigilancia rutinaria de los aislados resistentes a la vancomicina

Evaluation of antibacterial activity of essential oils extracted from Thymus satureioides and Mentha pulegium against antibiotic resistance bacteria from raw sheep milk

Aims@#This study evaluated in vitro resistance and susceptibility of Enterobacteriaceae (Escherichia coli and Klebsiella oxytoca strains) and Staphylococci strains, isolated from sheep’s milk, against antibiotics and essential oils from Thymus satureioides and Mentha pulegium.@*Methodology and results@#Antibiotic resistance tests were done using disc diffusion while essential oils were extracted by steam distillation and yields were calculated relative to plant dry matter. Gas chromatography-mass spectrometry (GC-MS) was used to analyze each oil's chemical composition. Amoxicillin + Clavulanic acid (AMC), Cefotaxime (CTX), Cefoxitine (FOX), Nalidixic acid (NA), Gentamicin (CN), Ciprofloxacin (CIP) and Ofloxacin (OFX) were very effective against the E. coli strains tested. Half of the strains were resistant to AMC, 60% to Ticarcillin (TIC) and 80% to Tetracycline (TE). Klebsiella oxytoca was resistant against AMC, FOX and TIC (100%). Antibiotic-resistant testing on Staphylococci strains indicated Staphylococcus capitis and S. chromogenes as the most sensitive. Staphylococcus aureus, S. xylosus and S. cohnii ureal exhibited less resistance to Oxacilin (OX), TE, Pristinamycin (PT), Erythromycin (E) and Penicillin (P). Mentha pulegium resulted in a higher yield of essential oil of 3.2% oil compared to T. satureioides with only 1.85% yield. The monoterpene oxygenated derivatives, monoterpene hydrocarbons and phenols are found in essential oil extracts. Thymus satureioides essential oil had high antibacterial activity even at low concentrations (0.2; 0.55 g/mL). The minimal bactericidal concentration (MBC) values indicate that the essential oils from the plants analyzed had bactericidal effects on all strains tested and are similar to the minimal inhibitory concentration (MIC) values.@*Conclusion, significance and impact of study@#The high antibacterial properties of these medicinal plants, against bacteria isolated from sheep’s milk, provide an opportunity to use these medicinal plants in the breeding sector, as additives and preservatives in the dairy industry.

Antibiotic resistance patterns of methicillin-resistant coagulase negative staphylococci isolated from blood cultures at a university hospital in Turkey

Aims@#Recent reports indicate that many coagulase-negative staphylococci (CoNS) strains are resistant to most antimicrobials used against staphylococcal infections. This study was aimed to determine the species distribution of the CoNS isolates in Lokman Hekim Ankara Hospital and determine their antimicrobial resistance characteristics.@*Methodology and results@#The study was conducted at Lokman Hekim University Ankara Hospital between February 2020 and August 2021. The 154 blood cultures included in the study were incubated in the BACTEC FX40 automated blood culture device. Identification and antimicrobial susceptibility tests of the samples with positive catalase tests were performed with the BD Phoenix Automated Microbiology Sensitivity System. The statistical significance level was accepted as p<0.05. Nineteen different types of methicillin-resistant coagulase-negative staphylococci (MRCoNS) isolated from different age groups were identified. Vancomycin resistance was observed in 20 samples (13%). Trimethoprim-sulfamethoxazole (TMP-SMX) resistance was seen in 44 of 152 samples (28.6%), linezolid resistance in 15 of 143 samples (10.5%) and daptomycin resistance in 16 of 146 blood samples (11%).@*Conclusion, significance and impact of study@#In our investigation, there has been a striking rise in the prevalence of vancomycin, TMP-SMX, linezolid and daptomycin resistance among infections with the MRCoNS. Identifying and classifying multidrug resistance on MRCoNS requires reliable epidemiological data to be collected and compared between healthcare facilities in different countries. The research finding reported in this paper will contribute to the determination of alternative antibiotics for treating MRCoNS.

Qualidade microbiológica da água e dos mexilhões Perna perna (Linnaeus, 1758) cultivados em Piúma, Espírito Santo, Brasil / Microbiological quality of water and Perna perna (Linnaeus, 1758) mussels cultivated in Piúma, Espírito Santo, Brazil

RESUMO Este estudo objetivou verificar a qualidade microbiológica da água e dos mexilhões cultivados pela Associação dos Maricultores de Piúma (AMPI), Espírito Santo, Brasil. Foram realizadas sete coletas de água e mexilhões, mensalmente, entre outubro de 2016 e maio de 2017. Os mexilhões foram coletados nos long lines da AMPI, e em cada mês foram coletados 40 mexilhões Perna perna e 100 mL de água do local. O material coletado foi destinado ao laboratório para a realização das análises microbiológicas em duplicata, número mais provável de coliformes totais (CT) e termotolerantes (Ctt), presença ou ausência de Salmonella sp e número de unidades formadoras de colônias de Staphylococcus aureus. Os resultados mostraram que o número de Ctt nas amostras de água no mês de janeiro estavam acima do permitido pela Resolução n° 357 do Conselho Nacional do Meio Ambiente (CONAMA). Já os níveis de Ctt e Staphylococcus aureus na carne dos mexilhões mostraram-se dentro do limite aceitável pela RDC n° 12 da Agência Nacional de Vigilância Sanitária (ANVISA). Entretanto, foram encontradas bactérias com características do gênero Salmonella sp nos meses de dezembro e março nos mexilhões, impossibilitando sua comercialização e seu consumo. Durante esses meses, a cidade tem alto fluxo de turistas. Por fim, recomenda-se a realização das análises microbiológicas continuamente, principalmente no período do verão, época que tem grande fluxo de turistas no município de Piúma e que registrou presença de Salmonella na carne dos mexilhões e níveis de Ctt na água acima do permitido pelas legislações vigentes.

ABSTRACT This study aimed to verify the microbiological quality of water and mussels cultivated by the Piúma Farmers Association (Associação dos Maricultores de Piúma — AMPI), Espírito Santo, Brazil. Seven samples of water and mussels were collected monthly from October 2016 to May 2017. The mussels were collected from the long lines of the AMPI, and in each month 40 mussels Perna perna and 100 mL of water were sampled from the site. The collected material was sent to the laboratory for duplicate microbiological analysis, Most Probable Number of Total (CT) and Thermotolerant (Ctt) coliforms, presence or absence of Salmonella sp and number of Staphylococcus aureus colony forming units. The results showed that the number of Ctt in the water samples in January was higher than that allowed by Resolution 357 of the National Environment Council (Conselho Nacional do Meio Ambiente — CONAMA). The levels of Ctt and Staphylococcus aureus in mussel meat were within the acceptable range by the Brazilian National Environment Council (Agência Nacional de Vigilância Sanitária — ANVISA) Resolution RDC No. 12. However, colonies with characteristics of Salmonella sp were found in December and March in the mussels, making it impossible to sell and consume. This period coincides with a high flow of tourists in the municipality. Finally, it is recommended to perform microbiological analyzes continuously, especially in the summer, where there are a lot of tourists in the city of Piúma, period that showed the presence of Salmonella in the meat of mussels and Ctt levels in water above the allowed current legislation.

Effect of donor-derived infection on clinical prognosis of kidney transplant recipients / 器官移植

Objective To evaluate the effect of donor-derived infection (DDI) on clinical prognosis of kidney transplant recipients. Methods Clinical data of 82 donors from donation after citizen's death and 148 kidney transplant recipients were retrospectively analyzed. According to the culture results of the lavage fluid of donor kidney, all recipients were divided into the lavage fluid culture of donor kidney positive group (positive group, n=92) and lavage fluid culture of donor kidney negative group (negative group, n=56). All recipients were assigned into the DDI group (n=19) and non-DDI group (n=129) according to whether they developed DDI or not. The distribution and composition ratio of positive strains in the lavage fluid of donor kidney were analyzed. The incidence of postoperative infection and other complications was assessed in the recipients. Perioperative conditions of the recipients were statistically compared between the DDI and non-DDI groups. The treatment efficacy and clinical prognosis of DDI recipients were evaluated. Results Among 148 recipients, 92 obtained positive culture results in the lavage fluid of donor kidney. A total of 131 pathogenic strains were isolated, including 41.2% (54/131) of Gram-positive cocci, 48.9% (64/131) of Gram-negative bacilli and 9.9%(13/131) of fungi. Among 148 recipients, 52 cases were infected. And 45% (41/92) and 20% (11/56) of the recipients were infected in the positive and negative group, respectively. Statistical significance was noted between two groups (P=0.002). Surgical site was the most common infection site in 52 infected recipients, followed by the urinary system. Nineteen recipients developed DDI with an incidence rate of 12.8% and fatality of 16%. Compared with the non-DDI recipients, DDI recipients had significantly higher graft loss rate and fatality, and longer postoperative hospital stay (all P < 0.05). Eight cases presented with carbapenem-resistant Klebsiella pneumoniae (CRKP) infection, after treatment with tigecycline and/or polymyxin and carbapenems, 3 cases died, and 3 underwent kidney graft resection. In the other 8 recipients with CRKP infection, 2 cases were treated with ceftazidime-avibactam (CAZ-AVI) alone, 3 treated with CAZ-AVI combined with carbapenems, and 3 initially treated with tigecycline combined with carbapenems followed by CAZ-AVI for salvage treatment. After corresponding treatment, the recipients achieved long-term survival. Conclusions DDI may lead to severe complications, while early specific antibacterial treatment plays a positive role.

Phenotypic Characterization and Antibiotic Susceptibility Profile of Coagulase Positive Staphylococci from The External Surfaces of Hospital Cockroaches (Periplaneta americana)

A study to phenotypically characterize and determine the antibiogram of coagulase positive Staphylococci (CoPS) from the external surfaces of hospital cockroaches (Periplaneta americana) was conducted using standard microbiological methods. Out of the 50 cockroaches collected from various hospitals in Uyo, sixty-two percent (n = 31) had coagulase positive Staphylococci which consisted of Staphylococcus aureus (44.0 %; n = 22) and Staphylococcus intermedius (18.0 %; n = 9). The CoPS isolates showed 100% resistance to Penicillin, Tetracycline, Clindamycin and 80.6% sensitivity to Amoxicillin-clavulanate. The CoPS showed multiple antibiotic resistances to ≥ 3 antibiotics, with 60 % exhibiting resistance to 6 antibiotics. Out of the 80 % (n = 31) of the multidrug resistant CoPS that were sensitive to Amoxicillin-clavulanate, none of them showed production of beta lactamase. The cockroaches bore multiple antibiotic resistant CoPS on their external surfaces and their contact can initiate contamination of patients' food. Pest control measures in hospital are hereby recommended to minimize cockroach related infections

Gangrenous mastitis in sheep caused by multidrug-resistant Staphylococcus haemolyticus / Mastite gangrenosa em ovinos causada por Staphylococcus haemolyticus multirresistente

Mastitis is a multifactorial disease and considered one of the most critical problems in the dairy industry worldwide. The condition is characterized by reduced milk and several abnormalities in the mammary gland. This study aimed to report an outbreak of gangrenous mastitis caused by multidrug-resistant Staphylococcus haemolyticus in a Santa Inês sheep herd. Eighteen sheep were affected, and five of them with severe clinical pictures were examined. The clinical and pathological picture were variable and characterized by apathy, anorexia, emaciation, opaque and brittle hair, apparent and congested episcleral vessels, and hyperthermia. These ewes had enlarged, firm, and painful mammary glands. Macroscopically, these lesions consisted of severe gangrenous mastitis, and microscopically, the primary lesions consisted of necrosis, thrombosis, and fibrosis of the mammary parenchyma. Milk samples from one of the five severely affected ewes were collected and cultured under aerobic or microaerophilic incubation at 37°C for 24 hours on sheep blood agar. The obtained colonies were then submitted to MALDI-TOF for speciation. The colonies were also submitted to an antimicrobial susceptibility test, genotyping of virulence factors and resistance genes were also performed. The isolates showed antimicrobial multiresistance since they were resistant to seven out of 13 tested antibiotics. The isolates were also positive for two staphylococcal enterotoxigenic genes (sec and see) and fibronectin-binding protein B (fnbB).(AU)

A mastite é uma doença multifatorial e é considerada um dos problemas mais importantes na indústria de laticínios no mundo todo. A condição é caracterizada pela redução de leite e várias anormalidades na glândula mamária. O objetivo deste estudo foi relatar um surto de mastite gangrenosa causada por Staphylococcus haemolyticus multirresistente em um rebanho ovino Santa Inês. Dezoito ovelhas foram afetadas e cinco delas com quadro clínico severo foram examinadas. O quadro clínico-patológico era variável quanto a severidade e consistia em apatia, anorexia, magreza, pelos opacos e quebradiços e vasos episclerais aparentes e ingurgitados. As ovelhas apresentavam glândulas aumentadas, firmes e dolorosas. Macroscopicamente, as principais lesões consistiam em mastite gangrenosa e microscopicamente havia necrose do parênquima glandular, trombose e fibrose. Amostras de leite de uma das cinco ovelhas severamente afetadas foram coletadas e cultivadas sob incubação aeróbica ou microaerofílica a 37°C por 24 horas em ágar sangue de ovelha. As colônias obtidas foram então submetidas ao MALDI-TOF para especiação. Além disso, as colônias foram submetidas a um teste de suscetibilidade antimicrobiana e foi realizada a genotipagem de fatores de virulência e genes de resistência. Os isolados apresentaram multirresistência antimicrobiana por serem resistentes a sete dos 13 antibióticos testados. Os isolados também foram positivos para dois genes enterotoxigênicos estafilocócicos (sec e see) e proteína B de ligação à fibronectina (fnbB).(AU)

Detection of the mecA gene in Staphylococcus spp. isolated from the nasal cavity of pigs - Preliminary data / [Detecção do gene mecA em Staphylococcus spp. isolados da cavidade nasal de suínos - dados preliminares]

O objetivo deste estudo foi realizar a caracterização genética de Staphylococcus spp. resistentes a meticilina isolados de suínos. Foram coletadas 30 amostras de swab nasal de suínos, abatidos em um frigorífico com Serviço de Inspeção Federal. Os isolados foram submetidos a análises macro e microscópicas que, em seguida, para detectar a resistência bacteriana, foram submetidos a ensaios fenotípicos da sensibilidade aos antimicrobianos. Posteriormente, as amostras resistentes a oxacilina, foram submetidas à reação em cadeia pela polimerase (PCR) para verificar a presença do gene mecA. Das 30 amostras analisadas, foram isolados 12 (40%) Staphylococcus spp. coagulase positiva, e 18 (60%) coagulase negativa, e, dentre os isolados, 26 (86,66%) foram resistentes a oxacilina sendo possível detectar o gene mecA em seis (23%) amostras. Este estudo evidencia a presença de genes de resistência em microrganismos comuns a microbiota de animais de produção que podem ser transmitidos ao homem. Além de chamar a atenção para a frequência e quantidade de antimicrobianos aos quais estes animais são expostos durante toda sua vida, podendo ser considerado um problema para a saúde única.(AU)

Detection of icaA, icaD, and bap genes and biofilm production in Staphylococcus aureus and non-aureus staphylococci isolated from subclinical and clinical bovine mastitis / Detecção dos genes icaA, icaD e bap e produção de biofilme por Staphylococcus aureus e estafilococos não-aureus isolados de mastite clínica e subclínica bovina

Algumas espécies de Staphylococcus causam infecções crônicas intramamárias e podem levar à formação de biofilme. No presente estudo, levantou-se a hipótese de que as espécies de Staphylococcus isolados da mastite bovina são capazes de formar biofilme in vitro associado à presença dos genes icaA, icaD ou bap. Um total de 200 isolados de Staphylococcus, sendo 100 Staphylococcus aureus de casos de mastite subclínica e 100 estafilococos não aureus (ENA) de casos de mastite subclínica e clínica, obtidos em duas fazendas leiteiras, no estado de São Paulo, foram avaliados quanto à capacidade de produzir biofilmes in vitro. A presença de icaA, icaD e bap foi confirmada por PCR, e a produção de biofilme em ágar vermelho congo (Congo Red Agar - CRA) e em teste de microplaca (Microtiter Plate - MtP) foi avaliada nos isolados de S. aureus e ENA. Os resultados mostraram a presença dos genes icaA, icaD e bap em S. aureus, mas não em ENA. A produção de biofilme pode estar associada à presença de outros fatores ou genes que estimulam a produção de biofilme in vitro. O ensaio de MtP serve como um modelo quantitativo para o estudo da aderência de espécies de estafilococos associados à mastite bovina.(AU)

Species distribution and clinical profiles of coagulasenegative staphylococci (CoNS) isolated from blood cultures among paediatric patients in Hospital Kuala Lumpur

@#Introduction: Coagulase-negative staphylococci (CoNS) is often considered as a culture contaminants but it can potentially be pathogenic to patients with risk factors. A combination of species identification and clinical criteria has been suggested in determining true CoNS bacteraemia. Objectives: To identify the species distribution, antibiotic susceptibility patterns and clinical profiles of CoNS isolated from blood cultures among paediatric patients in Hospital Kuala Lumpur (HKL). Methods: This study involved CoNS isolation from blood cultures of paediatric in-patients of the Paediatric Institute HKL. Isolates were identified to species level using Analytical Profile Index Staph identification strips and antimicrobial susceptibility pattern following Kirby-Bauer Disc Diffusion method. The clinical profiles of patients were obtained from their medical records. Results: Eleven CoNS species were identified from 148 isolates. Staphylococcus epidermidis was the most frequent species isolated (67.6%). The majority of the isolates showed resistance to penicillin (85.8%); while 70.3% were methicillin-resistant (MR) CoNS, which demonstrated a significant association with true infection (p=0.021). Predictors for significant CoNS infection included thrombocytopaenia, presence of predisposing factors, nosocomial infection, blood collected from peripheral vein, and CoNS isolated from two consecutive blood cultures. The most common predisposing factors for the isolation of CoNS were the presence of peripheral (54.1%) and central venous catheters (35.1%). Conclusion: CoNS can cause significant bloodstream infections. The isolation of CoNS from blood cultures should be carefully interpreted by considering the predictive factors. Local data regarding predictive factors of patients with culture-positive CoNS, species distribution and antimicrobial susceptibility pattern are useful to determine the significance of blood culture results and care management of patients

Medically Important Coagulase Negative Staphylococci

In recent years, coagulase negative Staphylococci(CONS) are increasingly recognized as one of the major causes of health care associated infections (HCAI). CONS infections are one of the major causes of morbidity and mortality in hospitalized patients. As CONS colonize various parts of the skin and mucous membranes, most of infections are en-dogenous in origin. However, they are also transmitted exogenously via various medical/ surgical procedures. The aim of present review article is to update and summarize the information available on medically important CONS with special reference to epidemiology, laboratory diagnosis and antibiotic resistance.

Neonatal sepsis due to coagulase negative Staphylococci: a study from Kashmir valley, India

Background: Neonatal sepsis is a leading cause of neonatal mortality and morbidity. Early diagnosis and treatment with appropriate antibiotics are important to improve the prognosis of neonatal sepsis. Coagulase-negative Staphylococci (CoNS) have emerged as prominent pathogens in the neonatal intensive care unit. These infections are rarely fatal, but they cause significant morbidity, especially among very low birth weight infants. This study was done to know the prevalence of Coagulase-negative Staphylococci in neonatal sepsis and to determine their antibiotic susceptibility pattern.Methods: A prospective study was conducted on blood samples of suspected neonatal septicaemia between August 2017 and May 2018 received at Department of Microbiology, Government Medical College, Srinagar. Blood culture was done by automated blood culture system, (BacT/Alert) and identification and antibiotic susceptibility was done by VITEK2 method.Results: Out of 356 neonates screened, there were 185 (53.4%) positive blood cultures. Among the culture positive cases, 107 (57.83%) were male and 78 (42.16%) were female. Early Onset Septicaemia cases (130 [70.27%]) were found to be three times higher than late onset Septicaemia (55 [29.72%]). Coagulase-negative Staphylococci (CoNs) (30.27%) were the most common organisms isolated followed by Acinetobacter sp (15.1%), Klebsiella sp (5.4%) S. aureus (4.8%) and E. coli (4.8%). All the isolates of CoNS were sensitive to linezolid and vancomycin and tigecycline. Methicillin resistance was seen in 84% isolates.Conclusions: Present study highlights the emergence of Coagulase-negative Staphylococci (CoNS) as predominant cause of neonatal septicaemia. Most of the isolates were resistant to methicillin which is alarming and a cause for concern.

Comparison of antibiotic resistance spectrum between methicillin-resistant and methicillin-susceptible coagulase-negative staphylococci nasal isolates among 1 001 HIV infectors / 中华疾病控制杂志

Objective We aimed to elucidate the prevalence and the antibiotic resistance spectrum of nasal coagulase-negative staphylococci (CoNS) colonization among HIV infectors in Guangzhou. Method After isolation and identification, all CoNS isolates were tested for the antibiotic susceptibility, and the antibiotic resistance genes. Result Among the 1 001 HIV infectors, the prevalence of CoNS and MRCoNS were 57.44% and 48.15%, respectively. The three predominant resistant antibiotics of MRCoNS isolates were penicillin, erythromycin and trimethoprim-sulfame thoxazole, while predominant detection rates of genes were Aac(6’)-aph(2’)、ermC and linA genes. The multidrug resistance rate of MRCoNS isolates were significantly higher than methicillin-susceptible coagulase-negative staphylococci (MSCoNS) isolates (80.69% versus 39.66%, P<0.001, OR=6.36). Conclusions The prevalence and multidrug resistant rates of nasal colonization CoNS and MRCoNS are high among HIV infectors in Guangzhou. MRCoNS isolates were 6.36 times more likely to be of multidrug resistance than MSCoNS isolates.

Isolation of Salmonella Typhimurium, Listeria monocytogenes and coagulase-positive Staphylococcus from salami sold at street fairs in Porto Alegre, Brazil / Isolamento de Salmonella Typhimurium, Listeria monocytogenes e Staphylococcus coagulase positiva de salames comercializados em feiras livres de Porto Alegre, Brasil

Salami is a ready-to-eat (RTE) product frequently purchased at street fairs in Porto Alegre. Salmonella enterica, Listeria monocytogenes, and coagulase-positive Staphylococcus (CPS) are important causes of foodborne disease and can be transmitted through the consumption of RTE foods. The aim of this study was to evaluate the presence of these pathogens in salami sold at street fairs. Ninety salami samples from three commercial brands available at street fairs were analyzed by routine bacteriological methods for Salmonella spp. and Listeria spp., as well as enumeration of CPS. In addition, two samples from each commercial brand were analyzed for water activity (aw). Samples of brand A showed aw values (0.938 and 0.942) above those set by the legislation, while brand B (0.849 and 0.860) and brand C (0.826 and 0.854) were compliant. Microbiological analyses showed that 67.7% were negative to all investigated bacteria. Salmonella Typhimurium was isolated from 4.4% (4/90) of salami samples, all from commercial brand A. ­Listeria monocytogenes was detected in 3.3% (3/90) of samples, from commercial brands B and C. Moreover, 7.7% (7/90) of samples contained CPS populations non-compliant with legislation. Although the great majority of salami sold at street fairs of Porto Alegre was compliant with standards, S. enterica, L. monocytogenes, and CPS ≥ 5 × 103 cfu.g-1 could be found in this RTE product. Therefore, control measures in the processing industry and consumer's education about foodborne illness prevention should be maintained.(AU)

Salame é um alimento pronto para o consumo frequentemente adquirido pela população em feiras livres de Porto Alegre. Salmonella enterica, Listeria monocytogenes e Staphylococcus coagulase positiva são importantes causas de doenças transmitidas por alimentos e podem ser veiculadas por alimentos prontos para o consumo. O objetivo desse estudo foi avaliar a presença desses patógenos em salames vendidos em feiras livres. Noventa amostras de salame pertencentes a três marcas comerciais foram analisados por métodos bacteriológicos de rotina quanto à presença de Salmonella spp. e Listeria spp., bem como enumeração de Staphylococcus coagulase positiva (SCP). Além disso, foi determinada a Atividade de Água (aw) de duas amostras de cada marca comercial. Amostras da marca A apresentaram valores de aw (0,938 e 0,942) acima do permitido na legislação, enquanto as amostras da marca B (0,849 e 0,860) e C (0,826 e 0,854) não violaram esse parâmetro. A análise microbiológica demonstrou que 67,7% das amostras foram negativas para todas as bactérias investigadas. Salmonella Typhimurium foi isolada de 4,4% (4/90) das amostras de salame, todas da marca comercial A. Listeria monocytogenes foi detectada em 3,3% (3/90) das amostras das marcas B e C. Além disso, 7,7% (7/90) das amostras apresentaram SCP acima do número permitido pela legislação. Apesar da grande maioria dos salames comercializados em feiras livres estarem de acordo com a legislação, S. enterica, L. monocytogenes e SCP ≥ 5 × 103 cfu.g-1 podem estar presentes nesse alimento pronto para o consumo. Dessa forma, o controle nas indústrias e a educação dos consumidores sobre a prevenção de doenças transmitidas por alimentos devem ser mantidos.(AU)

Occurrence of II and V staphylococcal cassette chromosome mec types among coagulase-negative staphylococci from Northeastern part of India

Introduction: Coagulase Negative Staphylococci, the most commonly isolated pathogen are becoming emerging threats to the community as well as to the nosocomial environment. The present study underscores the distribution of Staphylococcal cassette chromosome mec (SCCmec) types among Methicillin resistant Coagulase Negative Staphylococci from the environmental origin. Methods and Materials: Environmental and food sample (n = 460) from different location of northeastern region of India were collected for a period of one year and were phenotypically and genotypically screened using cefoxitin disc and PCR techniques for mecA and mecC gene detection. All the MR-CoNS isolates possessing mecA gene were subjected to 16srDNA sequencing for species identification. SCCmec typing was determined by evaluating using primer sets from type I to type V. Antibiotic susceptibility testing was performed for all the isolates. Statistical analysis with chi-square test using SPSS-21 statistical software. Results: Methicillin resistance shown by one hundred forty three isolates were carried out for molecular analysis, among them 53.84% serves as mecA carrier. Distribution of Staphylococcus haemolyticus was more frequent and was found that SCCmec types II and V were predominant among the study isolates. Linezolid was the drug of choice for the CoNS isolates. Statistical analysis showed an insignificant result for the tested antibiotics and SCCmec types. Conclusion: This study therefore interprets the relative importance of SCCmec types among MR-CoNS isolates.

Assessment of different methods for the detection of biofilm production in coagulase-negative staphylococci isolated from blood cultures of newborns

Abstract INTRODUCTION: Coagulase-negative staphylococci (CoNS) are a frequent cause of bacteremia, especially in neonates. The major virulence determinant in CoNS is the ability to produce biofilms, which is conferred by the icaADBC genes. This study aimed to assess different methods for the detection of biofilm formation in 176 CoNS isolates from blood cultures of newborns. METHODS: The presence of the icaACD genes was assessed by polymerase chain reaction (PCR), and biofilm formation was assessed on congo red agar (CRA), by the tube method (TM), and on tissue culture plates (TCP). RESULTS: Of the 176 CoNS isolates, 30.1% expressed icaACD and 11.4% expressed icaAD. The CRA assay and TM showed that 42% and 38.6% of the isolates were biofilm producing, respectively. On TCP, 40.9% of the isolates produced biofilms; 21% were weakly adherent and 19.9% were strongly adherent. When compared to the gold standard technique (PCR), the CRAassay showed 79% sensitivity and 84% specificity (kappa = 0.64), TM showed 78% sensitivity and 89% specificity (kappa = 0.68), and TCP showed 99% sensitivity and 100% specificity (kappa = 0.99). CONCLUSIONS: In this study, ~42% of CoNS isolates produced biofilms, and the presence of icaACD was associated with a greater capacity to form biofilms. Compared to the other phenotypic methodologies, TCP is an ideal procedure for routine laboratory use.

Catarrhal mastitis by Staphylococcus simulans in a nulliparous goat / Mastite catarral causada por Staphylococcus simulans em uma cabra nulipara

The present paper is a case report of a one-year old nulliparous Alpine Goat belonging to a dairy goat farm in semi-arid region of Brazil. Both glands were naturally infected by α-hemolytic Staphylococcus simulans and evolved similar clinical signs. The mammary glands presented an acute catarrhal mastitis with systemic clinical signs that responded positively to treatment with gentamicin associated with amoxicillin. The present report suggests the importance of the pathogenic potential of non-aureus Staphylococci strains (NAS) as a cause of clinical mastitis also in nulliparous animals. The isolate showed resistance to tetracycline and contained staphylococcal toxin production genes (sec, sec and TSST-1). Moreover, it has been reported that Staphylococcus simulans is an emerging pathogen in humans causing cutaneous and osteoarticular infections, mainly in those in close contact with farm animals. To the best of our knowledge, this is the first report of a clinical mastitis in a nulliparous goat caused by Staphylococcus simulans(AU)

O presente trabalho é o relato do caso de uma cabra nulípara da raça Parda Alpina, de um ano de idade, pertencente ao Setor de Caprinocultura da Universidade Federal da Paraíba ­ Bananeiras - Brasil. Ambas as glândulas foram naturalmente infectadas por Staphylococcus simulans α-hemolítico. As glândulas mamárias apresentaram mastite aguda catarral com envolvimento sistêmico, respondendo positivamente ao tratamento sistêmico com gentamicina associada a amoxicilina. O presente relato sugere a importância do potencial patogênico de Staphylococcus não-aureus (SNA) como causador de mastite clínica também em animais nulíparos. O isolado mostrou resistência a tetraciclina e continha genes de produção de toxinas estafilocócicas (sec, seg e TSST-1). Além disso, tem sido relatado que Staphylococcus simulans é um patógeno emergente em seres humanos causando infecções cutâneas e osteoarticulares, principalmente naqueles que têm contato íntimo com animais de fazenda. Até onde sabemos, este é o primeiro relato de uma mastite clínica em uma cabra nulípara causada por Staphylococcus simulans.(AU)