ABSTRACT
Brachyspira (B.) hyodysenteriae is a causative agent of swine dysentery that is responsible for death and economic losses in the pig industry. It is imperative that clinical samples be delivered fresh for accurate diagnosis. The viability and DNA detection of B. hyodysenteriae using lab-made (phosphate buffered saline and modified tryptic soy broth) or commercial transport media (C, D, and E) were compared by culturing and real-time PCR at 4degrees C or room temperature (RT), respectively. B. hyodysenteriae grown in D (Anaerobe Systems, USA) and E (Starplex Scientific, Canada) media was viable for 4 days at 4degrees C and RT. However, B. hyodysenteriae in A, B, and C (culture swab; BD Biosciences, USA) media were not recovered after 2 days at RT. Ct values for real-time PCR at 4degrees C and RT ranged from 27.2 +/- 2.1 (C) to 29.6 +/- 0.5 (B), and 28.0 +/- 0.9 (E) to 30.2 +/- 1.5 (B), respectively. Considering the field conditions, it is important that transport media is used for specimen isolation and PCR to obtain an accurate diagnosis of swine dysentery.
Subject(s)
Brachyspira , Brachyspira hyodysenteriae , Diagnosis , DNA , Dysentery , Polymerase Chain Reaction , Real-Time Polymerase Chain Reaction , SwineABSTRACT
Background: H1N1 infection is a viral infection and it is communicable from an infected person to the susceptible contacts. A large number of H1N1 cases and deaths had been reported in districts of western Rajasthan during the pandemic of H1N1 influenza in the year 2009-10. In 2012-2013 also, Influenza ‘A’ H1N1 virus once again has reappeared in western Rajasthan. Aims & Objective: To review deaths due to H1N1 Positive patients in Hospitals attached with Medical College. Material and Methods: The present study was a hospital based study. The study was conducted by taking information from H1N1positive patients and their relatives from Umed, M.G., M.D.M., and Chest & T. B. Hospitals of Jodhpur from 1st January, 2012 to 20th March, 2013. During our study period, 1877 samples were collected from the suspected Outdoor and Indoor patients of hospitals. The collected samples were transported in VTM (viral Transport Media) to the testing Laboratories of Medical College/ Desert Medicine Research Centre (DMRC), Jodhpur and subjected to RT-PCR technique for the detection of H1N1 Influenza virus. Results: Out of 1877 samples collected during our study period, 375 samples were found positive. The study shows an H1N1 case positivity rate of 19.97%. During the study period, 68 patients died of H1 N1 infection with a Case fatality rate of 18.13%. Out of 68 H1N1 positive patients’ deaths, deaths were more (45; 66.16%) in female than in male (23; 33.82%). Out of 45 women who died of H1N1 infection, 33.82% (23) of the women were pregnant. The study also reveals a higher incidence of H1N1 infection and mortality in younger age group. Conclusion: The Influenza A (H1N1) virus is still present two years after year 2009-10 pandemic. It has become a ubiquitous virus in the districts of Western Rajasthan region of India. In the event of new influenza outbreaks, hygienic and containment measures must be quickly and correctly implemented in order to avoid an epidemic.
ABSTRACT
Three comparative assays were performed seeking to improve the sensitivity of the diagnosis of Bordetella bronchiseptica infection analyzing swine nasal swabs. An initial assay compared the recovery of B. bronchiseptica from swabs simultaneously inoculated with B. bronchiseptica and some interfering bacteria, immersed into three transport formulations (Amies with charcoal, trypticase soy broth and phosphate buffer according to Soerensen supplemented with 5 percent of bovine fetal serum) and submitted to different temperatures (10ºC and 27ºC) and periods of incubation (24, 72 and 120 hours). A subsequent assay compared three selective media (MacConkey agar, modified selective medium G20G and a ceftiofur medium) for their recovery capabilities from clinical specimens. One last assay compared the polymerase chain reaction to the three selective media. In the first assay, the recovery of B. bronchiseptica from transport systems was better at 27ºC and the three formulations had good performances at this temperature, but the collection of qualitative and quantitative analysis indicated the advantage of Amies medium for nasal swabs transportation. The second assay indicated that MacConkey agar and modified G20G had similar results and were superior to the ceftiofur medium. In the final assay, polymerase chain reaction presented superior capability of B. bronchiseptica detection to culture procedures.
Três ensaios comparativos foram feitos com o objetivo de aperfeiçoar a sensibilidade do diagnóstico da infecção pela Bordetella bronchiseptica a partir de suabes nasais de leitões. O experimento inicial comparou a recuperação de B. bronchiseptica a partir de suabes, simultaneamente inoculados com B. bronchiseptica e algumas bactérias interferentes, imersos em três formulações para transporte (meio Amies com carvão, caldo tripticaseína de soja e tampão de fosfatos segundo Soerensen suplementado com 5 por cento de soro fetal bovino) e submetidos a diferentes temperaturas (10ºC e 27ºC) e períodos de incubação (24, 72 e 120 horas). O experimento subseqüente comparou três meios seletivos (ágar MacConkey, meio seletivo G20G modificado e o meio de ceftiofur) em relação às suas capacidades de recuperação a partir de amostras clínicas. O último experimento comparou a reação em cadeia pela polimerase aos três meios seletivos. No primeiro experimento, a recuperação de B. bronchiseptica nos sistemas de transporte foi melhor a 27ºC e as três formulações tiveram boas performances nesta temperatura, mas o conjunto das análises qualitativa e quantitativa apontou para o uso preferencial do meio Amies para o transporte de suabes nasais. O segundo experimento indicou que o ágar MacConkey e o G20G modificado mostraram resultados similares e foram superiores ao meio de ceftiofur. No experimento final, a técnica de reação em cadeia pela polimerase apresentou uma capacidade de detecção da B. bronchiseptica superior a do cultivo.