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1.
Pathogens ; 13(2)2024 Jan 26.
Artículo en Inglés | MEDLINE | ID: mdl-38392846

RESUMEN

In this new methodology, plasmonic ELISA (pELISA) was used to detect Circovirus porcine2 (PCV2) in serum samples without the need for plate reading equipment. This process occurs by adapting the conventional ELISA test with gold nanoparticles (AuNPs) to promote a color change on the plate and quickly identify this difference with the naked eye, generating a dark purple-gray hue when the samples are positive and red when the samples are negative. The technique demonstrated high efficiency in detecting samples with a viral load ≥ 5 log10 copies/mL. Plasmonic ELISA offers user-friendly, cost-effective, and reliable characteristics, making it a valuable tool for PCV2 diagnosis and potentially adaptable for other pathogen detection applications.

2.
Microorganisms ; 12(1)2024 Jan 02.
Artículo en Inglés | MEDLINE | ID: mdl-38257919

RESUMEN

Brazilian descendants of former Black-slave (quilombola) communities have been predisposed to several zoonotic diseases due to social vulnerability, characterized by subsistence and close contact with livestock and companion animals. Accordingly, the present study has assessed anti-Coxiella burnetii antibodies in 200 individuals and 20 dogs from four quilombola communities located in Paraná State, southern Brazil. Serum samples were tested by indirect immunofluorescence assay (IFA) using in-house and commercial diagnostic protocols, with analysis of seropositive titers and antibody type. Fisher's exact test was used to compare seropositivity to C. burnetti with binary variables, with variables with three or more possible responses submitted to logistic regression. In total, 44/200 (22%; 95% CI 16.82-28.24) people tested positive, and 4.5% had titers higher than 128, indicating a recent onset of C. burnetii infection. Seropositive individuals were statistically associated with the Limitão community (p = 0.0013), urban workers as occupations (p = 0.0475), consumption of undercooked meat (p = 0.0159), and contact with animal abortion (p = 0.0276). No seropositivity association was found for age, sex, education, habit of entering forest areas, consumption of game meat, consumption of raw milk, flea and tick bites, dog contact, or history of female miscarriage. Only one of 20 dogs was seropositive with a titer of 128, probably related to an acute animal infection. Despite the prevalence here being higher than previous Brazilian reports, including with symptomatic populations, the results were within range for worldwide outbreaks and occupational risk populations. To the reader's knowledge, this is the first human survey of Q fever in southern Brazil and should be considered a warning for C. burnetii in vulnerable populations, particularly Quilombola communities.

3.
Food Res Int ; 173(Pt 2): 113422, 2023 11.
Artículo en Inglés | MEDLINE | ID: mdl-37803760

RESUMEN

This study aimed to evaluate the level of counting by indicator microorganisms, identify the microbial ecology, detect Listeria monocytogenes and Salmonella sp., and determine the presence of virulence genes and biofilm formation. A total of 480 samples were collected from the surfaces of the equipment and utensils using sterile swabs for the detection of L. monocytogenes and Salmonella sp. and counting mesophilic aerobes, Enterobacteriaceae, Escherichia coli, and Pseudomonas sp. The microbial ecology was evaluated by sequencing the 16S rRNA gene. Genes for virulence and biofilm formation were analyzed and adhesion capacity was evaluated for L. monocytogenes and Salmonella sp. The mesophilic aerobe count was the highest in the dairy processing facility, followed by the pork and poultry slaughterhouses. L. monocytogenes was detected in all facilities, with the highest detection in the pork slaughterhouse, followed by the poultry and dairy facilities. Salmonella sp. was only detected in the dairy. Isolates of L. monocytogenes and Salmonella sp. showed poor adhesion to polystyrene surfaces, virulence genes, and biofilm formation. The frequent contaminants in the slaughterhouses were Pseudomonas, Acinetobacter, and Aeromonas in poultry, Acinetobacter, Pseudomonas, and Brevundimonas in pork, and Pseudomonas, Kocuria, and Staphylococcus in dairy. Our results provide useful information to understand the microbiological risks associated with contamination.


Asunto(s)
Listeria monocytogenes , Carne de Cerdo , Carne Roja , Animales , Porcinos , Aves de Corral , Microbiología de Alimentos , Industria Lechera , Brasil , ARN Ribosómico 16S , Escherichia coli , Salmonella/genética
4.
Braz J Microbiol ; 54(4): 3041-3049, 2023 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-37668830

RESUMEN

The objective of this study was to evaluate the potential antimicrobial and antibiofilm effect of ginger essential oil (GEO) and 6-gingerol on a multispecies biofilm formed by Listeria monocytogenes, Salmonella Typhimurium, and Pseudomonas aeruginosa on a polypropylene surface. The minimum inhibitory concentration concentrations obtained for GEO were 100 and 50 mg/mL and for 6-gingerol 1.25 mg/mL. Sessile cell counts ranged within 5.35-7.35 log CFU/cm2 in the control biofilm, with the highest sessile growth at 72 h. GEO treatments acted on the total population regardless of concentration at 1 and 48 h. L. monocytogenes behaved similarly to the total population, showing GEO action at 1 h and keeping the same pattern at 48, 72, and 96 h. Better action on S. Typhimurium was obtained at times of 1, 72, and 96 h. P. aeruginosa showed logarithmic reduction only when treated with GEO 50 mg at 24 h. As for 6-gingerol, in general, there was no significant action (p > 0.05) on the evaluated sessile cells. GEO showed antimicrobial activity against L. monocytogenes, S. Typhimurium, and P. aeruginosa, acting as an inhibitor of biofilm formation. As for 6-gingerol, it was considered a possible antimicrobial agent but without efficacy during biofilm formation.


Asunto(s)
Antiinfecciosos , Listeria monocytogenes , Aceites Volátiles , Zingiber officinale , Salmonella typhimurium , Aceites Volátiles/farmacología , Pseudomonas aeruginosa , Recuento de Colonia Microbiana , Biopelículas , Antiinfecciosos/farmacología
5.
Braz J Microbiol ; 54(3): 2173-2182, 2023 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-37582950

RESUMEN

Salmonella is present in the poultry production chain and is a major challenge in terms of food safety and animal health. The early Salmonella detection is one of the main tools to control and prevent the transmission of this pathogen. Microbiological isolation and serotyping to identify and differentiate Salmonella serovars are laborious processes, time-consuming, and expensive. Therefore, molecular diagnostic methods can be rapid and efficient alternatives to the detection of this pathogen. Thus, the aim herein was to standardize and evaluate the use of loop-mediated isothermal amplification (LAMP) in comparison with real-time PCR (qPCR) for detection of Salmonella associated with a multiplex qPCR for simultaneous identification and differentiation of S. Enteritidis, S. Typhimurium, S. Pullorum, and S. Gallinarum. The LAMP, qPCR, and multiplex qPCR assays were comparable in specificity. The three techniques were evaluated for specificity for 16 different serovars of Salmonella and for 37 strains of the serovars of interest. The limit of detection and the efficiency of the LAMP, qPCR, and multiplex qPCR reactions were determined. The techniques were applied to 33 samples of chicken carcasses and compared to the results of conventional microbiology for validation. As results, LAMP was specific in the detection of different Salmonella serovars but presented lower limit of detection ranging from 101 to 104 CFU/reaction. In comparison, qPCR could detect less cells (100 to 102 CFU/reaction), reaching equal specificity and better repeatability in the assays. The qPCR multiplexing for identification of the different serovars also showed good specificity, with the detection threshold between entre 101 and 102 CFU/reaction. The results obtained in the analyses on poultry carcasses suggested a correspondence between the results obtained in molecular methods and in conventional microbiology. Thus, the proposed assays are promising for the diagnosis of Salmonella in poultry carcasses, already proved to be faster and more efficient than conventional diagnostics techniques, being of great interest for poultry production, animal, and public health.


Asunto(s)
Aves de Corral , Salmonella , Animales , Aves de Corral/microbiología , Serogrupo , Inocuidad de los Alimentos/métodos , Pollos/microbiología , Sensibilidad y Especificidad
6.
Viruses ; 15(4)2023 03 30.
Artículo en Inglés | MEDLINE | ID: mdl-37112869

RESUMEN

Brazil currently ranks second in absolute deaths by COVID-19, even though most of its population has completed the vaccination protocol. With the introduction of Omicron in late 2021, the number of COVID-19 cases soared once again in the country. We investigated in this work how lineages BA.1 and BA.2 entered and spread in the country by sequencing 2173 new SARS-CoV-2 genomes collected between October 2021 and April 2022 and analyzing them in addition to more than 18,000 publicly available sequences with phylodynamic methods. We registered that Omicron was present in Brazil as early as 16 November 2021 and by January 2022 was already more than 99% of samples. More importantly, we detected that Omicron has been mostly imported through the state of São Paulo, which in turn dispersed the lineages to other states and regions of Brazil. This knowledge can be used to implement more efficient non-pharmaceutical interventions against the introduction of new SARS-CoV variants focused on surveillance of airports and ground transportation.


Asunto(s)
COVID-19 , SARS-CoV-2 , Humanos , SARS-CoV-2/genética , Brasil/epidemiología , Transportes , Vacunación
7.
Biomolecules ; 13(3)2023 03 17.
Artículo en Inglés | MEDLINE | ID: mdl-36979487

RESUMEN

Mosquito females of the genus Mansonia (Blanchard) can be a nuisance to humans and animals since they are voraciously hematophagous and feed on the blood of a variety of vertebrates. Despite their relevance, there is a lack of investigation into the blood-feeding patterns of the Mansonia species. Knowledge of the host preference is crucial in establishing the public health importance of a mosquito species and its potential to be involved in the transmission dynamics of pathogens. Species that are primarily anthropophilic can be more effective in spreading vector-borne pathogens to humans. In this study, we used an Illumina Nextera sequencing protocol and the QIIME2 workflow to assess the diversity of DNA sequences extracted in the ingested blood of mosquito species to evaluate the overall and local host choices for three species: Ma. titillans, Ma. Amazonensis, and Ma. humeralis, in rural areas alongside the Madeira River in the vicinities of the Santo Antonio Energia (SAE) reservoir in the municipality of Porto Velho, Rondônia, Western Brazil. By performing our analysis pipeline, we have found that host diversity per collection site showed a significant heterogeneity across the sample sites. In addition, in rural areas, Ma. amazonensis present a high affinity for B. taurus, Ma. humeralis shows an overall preference for C. familiaris and B. taurus, but also H. sapiens and E. caballus in urban areas, and Ma. titillans showed more opportunistic behavior in rural areas, feeding on wild animals and G. gallus, though with an overall preference for H. sapiens.


Asunto(s)
Culicidae , Humanos , Animales , Femenino , Brasil , Mosquitos Vectores , Conducta Alimentaria , Salud Pública
8.
Commun Med (Lond) ; 2: 41, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-35603276

RESUMEN

Background: The emergence of the Brazilian variant of concern, Gamma lineage (P.1), impacted the epidemiological profile of COVID-19 cases due to its higher transmissibility rate and immune evasion ability. Methods: We sequenced 305 SARS-CoV-2 whole-genomes and performed phylogenetic analyses to identify introduction events and the circulating lineages. Additionally, we use epidemiological data of COVID-19 cases, severe cases, and deaths to measure the impact of vaccination coverage and mortality risk. Results: Here we show that Gamma introduction in São José do Rio Preto, São Paulo, Brazil, was followed by the displacement of seven circulating SARS-CoV-2 variants and a rapid increase in prevalence two months after its first detection in January 2021. Moreover, Gamma variant is associated with increased mortality risk and severity of COVID-19 cases in younger age groups, which corresponds to the unvaccinated population at the time. Conclusions: Our findings highlight the beneficial effects of vaccination indicated by a pronounced reduction of severe cases and deaths in immunized individuals, reinforcing the need for rapid and massive vaccination.

9.
PLoS Negl Trop Dis ; 16(5): e0010392, 2022 05.
Artículo en Inglés | MEDLINE | ID: mdl-35536865

RESUMEN

Q fever and brucellosis are zoonoses that cause fever and other systemic clinical signs in humans; their occurrences are neglected and the differential diagnosis for some diseases is disregarded. This study aimed to investigate the seropositivity for Coxiella burnetii and Brucella spp. antibodies in patients suspected of dengue from 38 municipalities in the state of São Paulo, Brazil. The samples (n = 604) were obtained by convenience from the Adolfo Lutz Institute serum bank. Sera were subjected to an indirect immunofluorescence assay (IFA) using in-house and commercial diagnostic protocols to evaluate C. burnetii positivity. For Brucella spp., sera were subjected to rapid plate serum agglutination with buffered acidified antigen (AAT), slow tube serum agglutination (SAL), and 2-mercaptoethanol (2-ME) techniques. Associations and statistical inferences of the results were performed by logistic regression according to the clinical and demographic variables collected from the patients. Statistical analyses were performed using Statistical Analysis Software (SAS) and associations were considered when p value was <0.05. In all, 129 patients showed positive results for Q fever, indicating a seropositivity of 21.4% (95% CI 18.15-24.85). Patients with 14-20 days of symptoms had 2.12 (95% CI 1.34-3.35) times more chances of being seropositive for Q fever than patients with 7-13 days, and patients with 21-27 days of fever had 2.62 (95% CI 1.27-5.41) times more chances of being seropositive for Q fever than patients with 7-13 days. For the other variables analyzed, there were no significant associations between the groups. No positivity for brucellosis was observed. This is the most comprehensive study of people seropositive for Q fever in São Paulo state and provides additional data for the medical community in Brazil. It is suggested that Q fever may be an important differential diagnosis of febrile illnesses in the region, demanding the government's attention and investment in health.


Asunto(s)
Brucelosis , Coxiella burnetii , Dengue , Fiebre Q , Animales , Anticuerpos Antibacterianos , Brasil/epidemiología , Brucelosis/complicaciones , Dengue/complicaciones , Dengue/diagnóstico , Dengue/epidemiología , Fiebre/etiología , Humanos , Fiebre Q/complicaciones , Fiebre Q/diagnóstico , Fiebre Q/epidemiología , Estudios Seroepidemiológicos
10.
J Virol Methods ; 300: 114414, 2022 02.
Artículo en Inglés | MEDLINE | ID: mdl-34896456

RESUMEN

COVID-19 has posed a worldwide public health challenge affecting millions of people in different countries. Rapid and efficient detection of SARS-CoV-2 is essential for pandemic control. Reverse Transcription quantitative PCR (RT-qPCR) of nasopharyngeal swabs is the gold standard method for the virus detection, but the high demand for tests has substantially increased the costs and reduced the availability of reagents, including genetic material purification kits. Thus, the present study aimed to compare two bead-based RNA extraction methods (an in-house and a commercial kit) from nasopharyngeal swabs and RT-qPCR detection of SARS-CoV-2. Twenty-five positive and five negative nasopharyngeal swab samples were subjected to extraction of nucleic acids using both methods in an automated platform. Both protocols revealed a high correlation between Cycle Quantifications (Cqs) (r = 0.99, p < 0.0001). In addition, the in-house kit was 89.5 % cheaper when compared to the mean cost of commercial RNA extraction kits. The results show that the in-house protocol is an affordable and reliable option for RNA extraction for SARS-CoV-2 detection from nasopharyngeal swabs.


Asunto(s)
COVID-19 , Prueba de COVID-19 , Humanos , Fenómenos Magnéticos , Nasofaringe , ARN Viral/genética , SARS-CoV-2 , Sensibilidad y Especificidad
11.
Front Public Health ; 9: 745310, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34660520

RESUMEN

The emergence of several SARS-CoV-2 lineages presenting adaptive mutations is a matter of concern worldwide due to their potential ability to increase transmission and/or evade the immune response. While performing epidemiological and genomic surveillance of SARS-CoV-2 in samples from Porto Ferreira-São Paulo-Brazil, we identified sequences classified by pangolin as B.1.1.28 harboring Spike L452R mutation, in the RBD region. Phylogenetic analysis revealed that these sequences grouped into a monophyletic branch, with others from Brazil, mainly from the state of São Paulo. The sequences had a set of 15 clade defining amino acid mutations, of which six were in the Spike protein. A new lineage was proposed to Pango and it was accepted and designated P.4. In samples from the city of Porto Ferreira, P.4 lineage has been increasing in frequency since it was first detected in March 2021, corresponding to 34.7% of the samples sequenced in June, the second in prevalence after P.1. Also, it is circulating in 30 cities from the state of São Paulo, and it was also detected in one sample from the state of Sergipe and two from the state of Rio de Janeiro. Further studies are needed to understand whether P.4 should be considered a new threat.


Asunto(s)
COVID-19 , SARS-CoV-2 , Brasil , Humanos , Mutación , Filogenia , Glicoproteína de la Espiga del Coronavirus/genética
12.
Microb Drug Resist ; 27(4): 562-570, 2021 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-32865485

RESUMEN

Salmonella spp. is a foodborne pathogen present in the pork production chain, leading to potential contamination of end products and causing salmonellosis cases and outbreaks worldwide. The emergence of multidrug-resistant (MDR) Salmonella spp., especially isolates obtained from animal origin food, is a global concern. This study aimed to isolate Salmonella from swine mesenteric lymph nodes (MLN) and to characterize the virulence and antibiotic resistance profiles. MLN samples were obtained from a swine slaughterhouse and subjected to Salmonella spp. isolation. Ten MLN samples were positive and 29 isolates were identified based on PCR (invA and ompC) and serotyping: Derby, Cerro, and Give. Pulsed-field gel electrophoresis allowed to group the isolates based on their serotypes, resulting in three major clusters. All isolates presented the virulence-related genes pefA, sipA, sopB, spaN, and pagC. Relatively high numbers of Salmonella spp. were resistant to neomycin, polymyxin B, ciprofloxacin, tetracycline, and nalidixic acid. Furthermore, 25 isolates presented simultaneous resistance to three or more antibiotic classes, being characterized as MDR. The obtained results confirmed the relevance of swine as reservoirs of Salmonella spp. in the pork production chain and demonstrated the MDR profiles of isolates. Proper control and surveillance are required to avoid the contamination of end products.


Asunto(s)
Antibacterianos/farmacología , Farmacorresistencia Bacteriana Múltiple/genética , Salmonella/efectos de los fármacos , Porcinos/microbiología , Animales , Brasil , Ganglios Linfáticos/microbiología , Pruebas de Sensibilidad Microbiana , Virulencia
13.
Artículo en Inglés | LILACS, VETINDEX | ID: biblio-1347965

RESUMEN

Pattern minas cheese is a product developed with pasteurized milk, fermented with mesophilic cultures, and with the final addition of rennet. This cheese undergoes an artisanal maturation process and possesses a firm shell of yellowish color and striking and acidic flavor. Our study objective was to evaluate the microbiological quality of pattern minas cheese. We collected 40 samples from two micro regions (Uberlândia and Patos de Minas) of the Triângulo Mineiro and Alto Paranaíba mesor regions of the State of Minas Gerais, Brazil. The microbiological test results were recorded as counts of enterobacteria, Escherichia coli, coliforms at 35°C, coagulase-positive Staphylococcus and Salmonella spp. In the Patos de Minas micro region, the results were 45%, 35%, 20%, and 20% higher than 103 CFU/g for the counts of enterobacteria, Escherichia coli, coliforms at 35°C, and Staphylococcus coagulase-positive, respectively. Five percent of the analyzed samples were positive for Salmonella spp. in the Uberlândia micro region. Based on the findings of the microbiota in the cheese analyzed from the micro regions (Uberlândia and Patos de Minas), we concluded that the hygiene conditions in the manufacturing, handling, transport, and storage stages were precarious, requiring the implementation of Good Manufacturing Practices (GMP) systems, including Hazard Analysis and Critical Control Points (HACCP).(AU)


O queijo minas padrão é um produto elaborado com leite pasteurizado, fermentado com culturas mesófilas e adição de coalho. Esse queijo passa por um processo de maturação artesanal, possui uma casca firme de cor amarelada e sabor ácido. O presente trabalho avaliou a qualidade microbiológica de queijo minas padrão comercializado em duas microrregiões (Uberlândia e Patos de Minas) da mesorregião do Triângulo Mineiro e Alto Paranaíba do estado de Minas Gerais, Brasil. Foram examinadas 40 amostras de queijo. Os ensaios microbiológicos foram contagens de enterobactérias, Escherichia coli, coliformes a 35 oC, Staphylococcus coagulase positiva e pesquisa de Salmonella spp. Na microrregião de Patos de Minas, os resultados foram de 45%, 35%, 20% e 20% superiores a 103 CFU/g para as contagens de enterobactérias, Escherichia coli, coliformes a 35oC e Staphylococcus coagulase positiva, respectivamente. Cinco por cento das amostras analisadas foram positivas à pesquisa de Salmonella spp. Considerando a microrregião analisada (Uberlândia e Patos de Minas), a conclusão obtida foi que na região estudada, as condições de higiene nas etapas de fabricação, manuseio, transporte e armazenamento do queijo minas padrão são precárias, sendo necessária a implementação de sistemas de Boas Práticas de Fabricação (GMP), incluindo Análise de Perigos e Pontos Críticos de Controle (HACCP).(AU)


Asunto(s)
Queso/análisis , Queso/microbiología , Higiene , Staphylococcus , Escherichia coli
14.
Prev Vet Med ; 179: 105024, 2020 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-32417637

RESUMEN

This study evaluated 250 animals from 25 different processing lots, processed in four slaughterhouses in São Paulo state, Brazil for the presence of Salmonella in the mesenteric lymph nodes (10 g sample of each animal) and characterized the antibiotics resistance profile, the Pulsed Field Gel Electrophoresis - PFGE and Multi Locus Sequence Typing - MLST profiles of selected strains. The pathogen was present in 36.4% (n = 91, CL 95% 30.4-43.4) of samples and 72% (n = 18, CL 95% 50.6-87.9%) of the analyzed lots. The main serovars were S. Typhimurium (n = 23), Salmonella enterica subsp. enterica 1.4,5,12:i:- (n = 17), followed by S. Infantis (n = 12) and S. Havana (n = 11). Twenty-eight strains (30%) were classified as other serovars. Sixty-eight percent of the strains were resistant to Streptomycin and tetracycline, followed by ampicillin and sulphonamides (62.6%), chloramphenicol (56.0%), trimethoprim-sulfamethoxazole (41.8%) and nalidixic acid (40.7%). The antibiotics with lower resistance rates were cephalothin and aztreonam (both with 3.3% resistant), and ceftriaxone and cefepime (both with 7.7%). Multidrug-resistant strains (MDR) accounted for 70.3% of the isolates. Eight strains were submitted to MLST: four S. Typhimurium and one S.1.4,5,12:i:-, all belonging to the ST 19, two Salmonella Infantis, belonging to the ST 32 and one S. Derby, belonging to ST 40. Twenty-one isolates with different antibiotics resistance profiles from the most prevalent serovars were selected for PFGE analysis. Serovar S. Typhimurium (n = 11) revealed 4 pulsotypes and 1 cluster and S. 1.4,5,12:i:- (n = 10) revealed 5 pulsotypes and 4 clusters. The high prevalence of the pathogen, with its high rates of antibiotics resistance and belonging to genetic groups that are often associated with disease in humans, shows that the production chain of pork is a potential source of infection in salmonellosis cases. Therefore, effective preventive measures for pathogen control are needed to reduce the risk of foodborne diseases.


Asunto(s)
Farmacorresistencia Microbiana , Ganglios Linfáticos/microbiología , Salmonelosis Animal/epidemiología , Salmonella/fisiología , Enfermedades de los Porcinos/epidemiología , Animales , Brasil/epidemiología , Electroforesis en Gel de Campo Pulsado/veterinaria , Tipificación de Secuencias Multilocus/veterinaria , Prevalencia , Salmonella/efectos de los fármacos , Salmonelosis Animal/microbiología , Sus scrofa , Porcinos , Enfermedades de los Porcinos/microbiología
15.
Cell Transplant ; 28(1): 47-54, 2019 01.
Artículo en Inglés | MEDLINE | ID: mdl-30369261

RESUMEN

Crush injuries in peripheral nerves are frequent and induce long-term disability with motor and sensory deficits. Due to axonal and myelin sheath disruptions, strategies for optimized axonal regeneration are needed. Multipotent mesenchymal stromal cells (MSC) are promising because of their anti-inflammatory properties and secretion of neurotrophins. The present study investigated the effect of canine adipose tissue MSC (Ad-MSC) transplantation in an experimental sciatic nerve crush injury. Wistar rats were divided into three groups: sham ( n = 8); Crush+PBS ( n = 8); Crush+MSC ( n = 8). Measurements of sciatic nerve functional index (SFI), muscle mass, and electromyography (EMG) were performed. Canine Ad-MSC showed mesodermal characteristics (CD34-, CD45-, CD44+, CD90+ and CD105+) and multipotentiality due to chondrogenic, adipogenic, and osteogenic differentiation. SFI during weeks 3 and 4 was significantly higher in the Crush+MSC group ( p < 0.001). During week 4, the EMG latency in the Crush+MSC groups had better near normality ( p < 0.05). The EMG amplitude showed results close to normality during week 4 in the Crush+MSC group ( p < 0.04). There were no statistical differences in muscle weight between the groups ( p > 0.05), but there was a tendency toward weight gain in the Crush+MSC groups. Better motor functional recovery after crush and perineural canine Ad-MSC transplantation was observed during week 2. This was maintained till week 4. In conclusion, the canine Ad-MSC transplantation showed early pro-regenerative effects between 2-4 weeks in the rat model of sciatic nerve crush injury.


Asunto(s)
Tejido Adiposo/citología , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/fisiología , Regeneración Nerviosa/fisiología , Traumatismos de los Nervios Periféricos/fisiopatología , Traumatismos de los Nervios Periféricos/terapia , Neuropatía Ciática/fisiopatología , Neuropatía Ciática/terapia , Tejido Adiposo/metabolismo , Animales , Tratamiento Basado en Trasplante de Células y Tejidos/métodos , Perros , Electromiografía , Vaina de Mielina/fisiología , Ratas
16.
Cryobiology ; 81: 94-100, 2018 04.
Artículo en Inglés | MEDLINE | ID: mdl-29470953

RESUMEN

The aim of this study was to investigate the impact of prolonged storage at 4 °C on survival of cat preantral follicles (PAFs) pre- and post-vitrification. Ovaries were obtained from 12 queens and transported at 4 ºC within 2-6 h. Parts of the ovaries were stored for an additional 24 h or 72 h. The ovarian cortex was dissected, analyzed for viability (neutral red - NR) and morphology (histology - HE and ultrastructural analysis by TEM) and vitrified. We used 2 mm biopsy punches to obtain equal size pieces as the experimental units. After NR assessment, each sample was fixed and embedded in paraffin for HE staining to determine the number of morphologically intact follicles. Another 2 mm piece of ovary was subjected to TEM. NR viability assessment and HE results showed a similar tendency with PAF survival postvitrification even after prolonged cooling at 24 h and 72 h. With TEM, integrity of mitochondria, plasma and basal membranes as well as the presence of pre-granulose cells of PAFs were documented postvitrification for the control group and 24 h prolonged storage group, but not after 72 h storage. Our results showed that cat PAFs can survive prolonged storage followed by vitrification. The described set of techniques are applicable towards creating a gamete bank for endangered feline species.


Asunto(s)
Criopreservación/veterinaria , Folículo Ovárico , Vitrificación , Animales , Gatos , Frío , Femenino , Bancos de Tejidos
17.
Ciênc. rural ; 47(2): 20151218, 2017. tab
Artículo en Inglés | LILACS | ID: biblio-828439

RESUMEN

ABSTRACT: Equine sport modalities influence the prevalence and predisposition of musculoskeletal injuries in horses. This study aimed to evaluate the prevalence of location and type of orthopedic injuries of horses undergoing various physical activities. Data from 116 horses of different breeds and ages was analyzed. Physical activities included dressage, racing, polo pony, jumping, work and western performance. All horses had history of orthopedic lameness diagnosed by radiographs and/or ultrasound scans. The effect of sport on the affected anatomical site and type of lesion was performed using Fisher's exact test. Desmitis was more prevalent in animals that performed western sports than in the working ones. The number of fractures was greater in racing and polo pony animals than in working horses. Stifle lesions were more prevalent in dressage horses than working horses and had lower occurrence of shoulder injuries than jumping horses. Hind limb tendon injuries were lower in jumping than in dressage and western horses. We conclude that there is a relationship between location and type of injury and physical activities. In racing horses there is a predominance of young animals and higher prevalence of orthopedic injuries from traumatic events such as tendonitis, desmitis and fractures. In physical activities that require longer training and that animals were used for longer periods, eg. jumping, polo pony, dressage and work, age-related degenerative, such as joint disease, were predominant. In western sport animals the most common lesion was desmitis. Regarding limbs, forelimb injures were more often observed in racing horses, polo pony, jumping and working animals; whereas, dressage and Western sports horses presented more injuries in hind limbs.


RESUMO: As modalidades esportivas influenciam a prevalência e predisposição de lesões musculoesqueléticas em cavalos. Objetivou-se avaliar a prevalência do local e tipo de lesões ortopédicas de cavalos submetidos a diversas atividades físicas. Foram avaliados 116 prontuários de equinos de adestramento, corrida, polo, salto, trabalho e Western, de raças e idades variadas, com histórico de claudicação e confirmação do diagnóstico por radiografia ou ultrassonografia da lesão ortopédica. O efeito da atividade física sobre o sítio anatômico afetado e tipo de lesão foi avaliado pelo teste Exato de Fisher. A desmite foi mais prevalente em animais de Western quando comparada com animais de trabalho. Fratura foi maior em Corrida e Polo comparado com Salto; e maior em Corrida comparado com Trabalho. As alterações fêmoro-tíbio-patelares foram mais prevalentes em adestramento do que trabalho e estes apresentaram menor ocorrência de lesões de articulação escápulo-umeral que os de salto. As lesões tendíneas de membros pélvicos foram menos prevalentes nos animais de salto do que nos de adestramento e Western. Conclui-se que há uma relação entre local e tipo de lesão com as atividades físicas. No turfe, há um predomínio de animais jovens e maior prevalência de lesões ortopédicas provenientes de eventos traumáticos, como tendinite, desmites e fraturas. Em atividades físicas que requerem um treinamento mais prolongado e que os animais são usados por mais tempo, como os de salto, polo, adestramento e trabalho, predominam lesões degenerativas relacionadas à idade, como doença articular. Em animais de Western, predominam as desmites. Quanto à porcentagem das injúrias ortopédicas distribuídas pelos membros, preponderam alterações em membros torácicos nos animais de turfe, polo, salto e trabalho e nos membros pélvicos nos animais de adestramento e de Western.

18.
Arq. Inst. Biol ; 84: e0792015, 2017. tab, graf
Artículo en Inglés | LILACS, VETINDEX | ID: biblio-887848

RESUMEN

The objective of this work was to verify the microbiological profile of different types of salads from hospital kitchens. During the period from 2010 to 2014, the Public Food Guidance Service (SOAP) received 641 samples of salads from two public hospitals in the Central West region of the São Paulo state, where they were submitted to microbiological analysis in order to determine the most probable number (MPN) of coliforms at 35 and 45ºC, carry out Salmonella spp. study and coagulase-positive staphylococci count. The results showed that in 30.56% of samples the coliform count at 35ºC was above 1,100 MPN/g and 12.17% of samples presented coliforms at 45ºC above 100 MPN/g, which is the maximum limit established by Brazilian law. The prevalence of contaminated samples among those without heat treatment was at 97.44%, and for samples with heat treatment the prevalence was at 2.56% for both cooked and braised foods. All samples were negative for Salmonella spp. presence and showed coagulase-positive staphylococci count at < 1.0 × 102 UFC/g. Although no pathogenic agents were found, the high count for indicator microorganisms in a large number of samples suggests that the practices of obtaining and manipulating these foods are inadequate, facilitating the risk of contamination with pathogens, including other agents not included in this research. Thus, food and nutrition units must pay attention to food preparation procedures, especially since these meals are served to indoor patients.(AU)


O objetivo deste trabalho foi verificar o perfil microbiológico de diferentes tipos de saladas provenientes de cozinhas hospitalares. No período de 2010 a 2014, o Serviço de Orientação à Alimentação Pública (SOAP) recebeu 641 amostras de saladas provenientes de dois hospitais públicos da região centro-oeste do estado de São Paulo, onde foram submetidas às análises microbiológicas para a determinação do número mais provável (NMP) de coliformes a 35 e 45ºC, pesquisa de Salmonella spp. e contagem de estafilococos coagulase positiva. Os resultados revelaram que em 30,56% das amostras a contagem de coliformes a 35ºC foi maior que 1.100 NMP/g, e 12,17% apresentaram coliformes a 45ºC acima de 100 NMP/g, limite máximo estabelecido pela legislação brasileira. A prevalência de amostras contaminadas sem tratamento térmico foi de 97,44% e de 2,56% para amostras com tratamento térmico, cozidas ou refogadas. Todas as amostras foram negativas para presença de Salmonella spp. e apresentaram contagem de estafilococos coagulase positiva < 1,0 × 102 UFC/g. Embora não tenham sido encontrados agentes patogênicos, as altas contagens de micro-organismos indicadores em grande parte das amostras sugerem que as práticas de obtenção e manipulação desses alimentos estão inadequadas, possibilitando risco de contaminação por patógenos, inclusive outros agentes não contemplados nesta pesquisa. Portanto, as unidades de alimentação e nutrição hospitalares devem se atentar quanto aos procedimentos de preparo de alimentos, sobretudo porque essas refeições são servidas a indivíduos hospitalizados.(AU)


Asunto(s)
Humanos , Plantas , Salmonella , Coliformes , Microbiología de Alimentos , Hospitales , Pacientes , Contaminación Ambiental
19.
Ciênc. rural ; 42(1): 131-135, 2012. tab
Artículo en Portugués | LILACS | ID: lil-612729

RESUMEN

A pesquisa de Salmonella em carcaças de aves tem mostrado resultados discrepantes, dependendo se as amostras foram colhidas ainda na indústria, imediatamente após o chiller ou no comércio varejista, quando se encontram submetidas à refrigeração por vários dias. Técnicas mais sensíveis, tais como a Reação em Cadeia pela Polimerase (PCR), podem fornecer dados importantes sobre o efeito do resfriamento sobre as células do patógeno, comparando seus resultados com os da metodologia microbiológica convencional (MC). Foram colhidas 130 carcaças de frango, sendo que 65 foram obtidas ainda na indústria (pós-chiller) e imediatamente analisadas e 65 carcaças embaladas e estocadas a 5°C por 72 horas (simulando o varejo), sendo então realizada a pesquisa do patógeno por ambas as técnicas. Do total analisado (130 amostras), a PCR foi capaz de detectar 58 positivas (44,6 por cento) e a MC 50 (38,5 por cento). Ambas as técnicas detectaram um número superior de amostras positivas para Salmonella em carcaças colhidas ainda na indústria, quando comparadas às do varejo. A PCR detectou 50,77 por cento de positividade em amostras da indústria e 38,46 por cento em amostras do varejo. Para a MC, esses valores foram de 46,15 por cento (indústria) e 30,77 por cento (varejo). Concluímos que o resfriamento das carcaças a 5°C por 72 horas pode ser um fator limitante na detecção de Salmonella quando a pesquisa do patógeno se faz pela metodologia microbiológica convencional.


The survey of Salmonella in poultry carcasses has shown conflicting results, depending on whether the samples were taken yet in the factory, immediately after the chiller or retail market, when they are subjected to refrigeration for several days. More sensitive techniques such as Polymerase Chain Reaction (PCR) can provide important data on the effect of cooling on the cells of the pathogen by comparing their results with those of conventional microbiological methods (CM). It was collected 130 chicken carcasses, wich 65 were obtained yet in the industry (post-chiller) and immediately analyzed and 65 carcasses packaged and stored at 5°C for 72 hours (simulating retail), and then research of the pathogen was performed by both techniques. Of the analyzed total (130 samples), PCR was able to detect 58 positive (44.6 percent) and CM, 50 (38.5 percent). Both techniques detected a higher number of samples positive for Salmonella on carcasses collected yet in the factory when compared to those of retail. The PCR detected 50.77 percent of positive industry samples and 38.46 percent of retail samples. CM for these values was 46.15 percent (factory) and 30.77 percent (retail). We conclude that the cooling of the carcasses for at 5°C for 72 hours can be a limiting factor in the detection of Salmonella when the pathogen research is done by conventional microbiological methods.

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