Polymerase chain reaction (PCR) for the detection of Salmonella in artificially inoculated chicken meat

The aim of this study was to develop a polymerase chain reaction (PCR) protocol for the detection of Salmonella in artificially contaminated chicken meat. Tests were performed with different dilutions of Salmonella Typhimurium or Salmonella Enteritidis cells (10-7, 10-8 or 10-9 CFU/mL) inoculated in chicken meat samples, in order to establish the limits of detection, incubation times (0, 6, 8 and 24 hours of pre-enrichment in PBW 1%) and three DNA extraction protocols (phenol-chloroform, thermal treatment and thermal treatment and Sephaglass). The assay was able to detect until 10-9 CFU/mL of initial dilution of Salmonella cells inoculated in chicken meat, which allows detection of Salmonella within 48 hours, including 24 hours of pre-enrichment and using the phenol-chloroform DNA extraction protocol. As the results are obtained in a shorter time period than that of microbiological culture, this procedure will be useful in the methodology for detection of Salmonella in chicken.

Milk flow, teat morphology and subclinical mastitis prevalence in Gir cows

The aim of this work was to evaluate the association between milk flow, teat morphological measurements and subclinical mastitis prevalence in Gir cows. Eighty cows in the 2nd and 3rd lactations, with 90 to 200 days of lactation, were divided according to milk flow during milking into fast or slow groups. Teat morphometry was assessed by ultrasound scanning of the right anterior teat and external measurements. Milk samples were collected for somatic cells count (SCC) and microbiological culture. The effect of milk flow during milking was evaluated by analysis of variance of milk yield, SCC, morphometry and external measurements. The association of morphometry and external measurements of the teats with the SCC and microorganisms found in milk were analysed. Milk flow was significantly correlated to milk production. Gir cows with slower milk flow had longer teat canal and greater milk yield, in comparison to cows with fast milk flow. Teat-end to floor distance influenced SCC of Gir cows. Prevalence of subclinical mastitis and the type of mastitis-causing pathogens were not affected by milk flow during milking

Etiology, antimicrobial susceptibility profile of Staphylococcus spp. and risk factors associated with bovine mastitis in the states of Bahia and Pernambuco

The purpose of this paper was to study the etiology of mastitis, determine the antimicrobial susceptibility profile of Staphylococcus spp. and to identify the risk factors associated with infection in dairy cows in the states of Bahia and Pernambuco, Brazil. From the 2,064 milk samples analyzed, 2.6% were associated with cases of clinical mastitis and 28.2% with subclinical mastitis. In the microbiological culture, Staphylococcus spp. (49.1%) and Corynebacterium spp. (35.3%) were the main agents found, followed by Prototheca spp. (4.6%) and Gram negative bacilli (3.6%). In the antimicrobial susceptibility testing, all 218 Staphylococcus spp. were susceptible to rifampicin and the least effective drug was amoxicillin (32.6%). Multidrug resistance to three or more drugs was observed in 65.6% of Staphylococcus spp. The risk factors identified for mastitis were the extensive production system, not providing feed supplements, teat drying process, not disinfecting the teats before and after milking, and inadequate hygiene habits of the milking workers. The presence of multiresistant isolates in bovine milk demonstrates the importance of the choice and appropriate use of antimicrobial agents. Prophylactic and control measures, including teat antisepsis and best practices for achieving hygienic milking should be established in order to prevent new cases of the disease in herds.

Suppurative intracranial processes in 15 domestic ruminants

In addition to listeriosis which is relatively common in ruminants, there are three other uncommon suppurative intracranial processes (SIP) identifiable in adult ungulates as brain abscess, basilar empyema and suppurative meningitis. The present paper reports the epidemiological, clinical, laboratorial, pathological and microbiological findings of 15 domestic ruminants with SIP. A total of 15 animals were selected (eight sheep, four cattle and three goats); with the definitive diagnoses of basilar empyema (n=3), brain abscess (n=1), listeriosis (n=5) and suppurative meningitis (n=6). Hematology revealed leukocytosis with inversion of the lymphocyte/ neutrophil ratio in 4 cases. In the majority of animals, cerebrospinal fluid (CSF) presented light yellow coloration and cloudy aspect due to neutrophilic pleocytosis (15 - 997 leukocytes/µL). Microbiological culture of CSF or central nervous system (CNS) fragments resulted on isolation of Trueperella (Arcanobacterium) pyogenes,Listeria monocytogenes,Escherichia coli and Stenotrophomonas sp. In a goat with thalamic abscess, microbiological assay was not performed, but Gram positive bacilli type bacteria were observed in histology. The diagnosis of these outbreaks was based on the association of epidemiological, clinical, pathological and bacteriological findings; reiterating that the infectious component remains an important cause of CNS disease in domestic ruminants and also shows the need for dissemination of information about the most effective preventive measures for the ranchers.

Risk factors associated with the antimicrobial resistance of Staphylococcus aureus isolated from bovine mastitis

The objective of this study was to evaluate herd management practices and mastitis treatment procedures as risk factors associated with Staphylococcus aureus antimicrobial resistance. For this study, 13 herds were selected to participate in the study to evaluate the association between their management practices and mastitis treatment procedures and in vitro antimicrobial susceptibility. A total of 1069 composite milk samples were collected aseptically from the selected cows in four different periods over two years. The samples were used for microbiological culturing of S. aureus isolates and evaluation of their antimicrobial susceptibility. A total of 756 samples (70.7%) were culture-positive, and S. aureus comprised 27.77% (n=210) of the isolates. The S. aureus isolates were tested using the disk-diffusion susceptibility assay with the following antimicrobials: ampicillin 10mg; clindamycin 2μg; penicillin 1mg; ceftiofur 30μg; gentamicin 10mg; sulfa-trimethoprim 25μg; enrofloxacin 5μg; sulfonamide 300μg; tetracycline 30μg; oxacillin 1mg; cephalothin 30μg and erythromycin 5μg. The variables that were significantly associated with S. aureus resistance were as follows: the treatment of clinical mastitis for ampicillin (OR=2.18), dry cow treatment for enrofloxacin (OR=2.11) and not sending milk samples for microbiological culture and susceptibility tests, for ampicillin (OR=2.57) and penicillin (OR=4.69). In conclusion, the identification of risk factors for S. aureus resistance against various mastitis antimicrobials is an important information that may help in practical recommendations for prudent use of antimicrobial in milk production.

Cutaneous infection caused by Corynebacterium pseudodiphtheriticum: a microbiological report

We report here a rare case of cutaneous infection due to Corynebacterium pseudodiphtheriticum. The patient presented to the clinical laboratory with a skin ulcer on his left leg. Gram-stained preparation of the purulent secretion revealed the presence of numerous rod-shaped Gram-positive organisms in the absence of any other species. The organism was grown in pure culture on sheep blood agar and was further identified as C. pseudodiphtheriticum using a commercial identification system (API-Coryne, BioMérieux, France). The infection was successfully treated with ciprofloxacin. This case emphasizes the importance of the clinical microbiology laboratory in correctly identifying Gram-positive organisms obtained in pure culture from skin ulcers.

Clinical and microbiological features of cryptococcal meningitis

Introduction In this study, the clinical features, underlying diseases and clinical outcomes of patients with cryptococcosis were investigated. In addition, a molecular analysis of the Cryptococcus neoformans species complex isolated from these patients was performed. Methods A prospective study of 62 cases of patients with cryptococcal infection was conducted at the Hospital de Doenças Tropicais de Goiás Dr. Anuar Auad from 2009-2010. Cryptococcal meningitis cases were diagnosed by direct examination and cerebrospinal fluid (CSF) sample culture. The profiling of these patients was assessed. The CSF samples were submitted to India ink preparation and cultured on Sabouraud dextrose agar, and C. neoformans was identified by the production of urease, a positive phenoloxidase test and assimilation of carbohydrates. C. neoformans and C. gattii isolates were distinguished by growth on L-canavanine-glycine-bromothymol blue medium, and molecular analysis was conducted via PCR fingerprinting reactions using M13 and (GACA)4 primers. Results From the 62 patients with cryptococcosis, 71 isolates of CSF were obtained; 67 (94.4%) isolates were identified as C. neoformans var. grubii/VNI, and 4 (5.6%) were identified as C. gattii/VGII. Of these patients, 53 had an HIV diagnosis. The incidence of cryptococcosis was higher among patients 20-40 years of age, with 74.2% of the cases reported in males. Cryptococcus-related mortality was noted in 48.4% of the patients, and the symptoms were altered sensorium, headache, fever and stiff neck. Conclusions The high morbidity and mortality observed among patients with cryptococcosis demonstrate the importance of obtaining information regarding the epidemiological profile and clinical course of the disease in the State of Goiás, Brazil.

The microbiological signature of human cutaneous leishmaniasis lesions exhibits restricted bacterial diversity compared to healthy skin

Localised cutaneous leishmaniasis (LCL) is the most common form of cutaneous leishmaniasis characterised by single or multiple painless chronic ulcers, which commonly presents with secondary bacterial infection. Previous culture-based studies have found staphylococci, streptococci, and opportunistic pathogenic bacteria in LCL lesions, but there have been no comparisons to normal skin. In addition, this approach has strong bias for determining bacterial composition. The present study tested the hypothesis that bacterial communities in LCL lesions differ from those found on healthy skin (HS). Using a high throughput amplicon sequencing approach, which allows for better populational evaluation due to greater depth coverage and the Quantitative Insights Into Microbial Ecology pipeline, we compared the microbiological signature of LCL lesions with that of contralateral HS from the same individuals.Streptococcus, Staphylococcus,Fusobacterium and other strict or facultative anaerobic bacteria composed the LCL microbiome. Aerobic and facultative anaerobic bacteria found in HS, including environmental bacteria, were significantly decreased in LCL lesions (p < 0.01). This paper presents the first comprehensive microbiome identification from LCL lesions with next generation sequence methodology and shows a marked reduction of bacterial diversity in the lesions.

Collection of tracheal aspirate: safety and microbiological concordance between two techniques

Objective: To evaluate the safety of the performance of the traditional and protected collection techniques of tracheal aspirate and to identify qualitative and quantitative agreement of the results of microbiological cultures between the techniques. Method: Clinical, prospective, comparative, single-blind research. The sample was composed of 54 patients of >18 years of age, undergoing invasive mechanical ventilation for a period of ≥48 hours and with suspected Ventilator Associated Pneumonia. The two techniques were implemented in the same patient, one immediately after the other, with an order of random execution, according to randomization by specialized software. Results: No significant events occurred oxygen desaturation, hemodynamic instability or tracheobronchial hemorrhage (p<0.05) and, although there were differences in some strains, there was qualitative and quantitative agreement between the techniques (p<0.001). Conclusion: Utilization of the protected technique provided no advantage over the traditional and execution of both techniques was safe for the patient.


Development of an indirect ELISA to detect Corynebacterium pseudotuberculosis specific antibodies in sheep employing T1 strain culture supernatant as antigen

Corynebacterium pseudotuberculosis is the etiologic agent of caseous lymphadenitis (CLA), a chronic disease that affects goats and sheep, characterized by granuloma formation in subcutaneous and internal lymph nodes. CLA causes significant economic losses to commercial goat herds. In this study, we aimed to test secreted antigens secreted from T1 strain bacteria grown in brain heart infusion (BHI) broth in an indirect ELISA system to determine the presence of specific immunoglobulins against C. pseudotuberculosis. We analyzed the BHI antigen electrophoretic profile and the recognition pattern by infected sheep sera samples. The ELISA results were compared with multiplex PCR assay and IFN-gamma production. The ELISA was able to discriminate between negative and positive animals, with a sensitivity of 89% and a specificity of 99%, using microbiological isolation as gold standard. When this assay was compared with multiplex PCR and specific IFN-gamma quantification, six discrepant results were found among thirty-two samples. We concluded that the ELISA using antigens secreted from C. pseudotuberculosis T1 strain growth in BHI broth culture can be used for the serodiagnosis of CLA in sheep.

Production of probiotic fresh white cheese using co-culture with Streptococcus thermophilus

In this research, the probiotic Streptococcus thermophilus was inoculated into milk as co-culture to produce probiotic cheese. The effects of using Streptococcus thermophilus with other probiotic bacteria on cheese composition, and microbiological viability during 28 days of storage were investigated. Sensorial properties were determined only at 1st and 28th days of storage. The results showed that the use of Streptococcus thermophilus as co-culture in probiotic cheese production did not affect negatively the cheese components. Fat and dry matter properties of cheese weren't influenced by added probiotic bacteria. However, different level of pH, salt and lactic acid were detected. All probiotic bacteria were present in high levels throughout storage of cheeses, above 7 Log cfu.g- 1, threshold required for probiotic activity. Sensory panel showed that the highest average sensory evaluation points were recorded in cheeses made with Streptococcus thermophilus plus Lactobacillus casei, whereas other probiotic bacteria combinations had been affected less in regard to taste or appearance.

Consumer Culture Theory (CCT) já é uma escola de pensamento em marketing?

A década de 1980 trouxe uma visão alternativa à corrente positivista predominante no campo de pesquisa do consumidor: a Consumer Culture Theory (CCT), que assume uma orientação epistemológica baseada no interpretativismo e na pesquisa qualitativa. Diante do destaque alcançado pela CCT, levantou-se a seguinte questão: a CCT já pode ser considerada uma escola de pensamento em marketing autônoma? Pautados em três critérios fundamentais para a qualificação de uma escola de pensamento (reconhecimento acadêmico, corpo de conhecimento e contribuições), foi realizada uma desk research, baseada em periódicos e artigos da área e na construção de um corpus de pesquisa construído com base nas referências contidas no texto seminal Consumer Culture Theory (CCT): twenty years of research. A conclusão é de que a CCT atende aos critérios adotados na presente pesquisa, podendo ser considerada uma escola de pensamento a utônoma dentro do campo de pesquisa do consumo.