Phenotypic and genotypic antibiotic resistance patterns in Salmonella Typhimurium and its monophasic variant from pigs in southern Spain.

The high incidence of human salmonellosis and multi-drug resistant (MDR) strains of Salmonella Typhimurium (ST) is of concern to global public and animal health. Our research, by means of the broth microdilution method, evaluated the Minimum Inhibitory Concentration (MIC) distribution of 12 antimicrobials against a collection of 73 ST and mST and S. typhimurium monophasic variant 4,[5],12:i:- (mST) isolates from slaughtered pigs reared in extensive systems in southern Spain, and also 12 resistance-associated genes or antimicrobial resistance (AMR) determinants using qPCR. Our data revealed that 98.6% of strains were MDR, with resistance to cephalothin/tetracycline/sulfamethoxazole-trimethoprim/ampicillin/chloramphenicol being the most common pattern (55.6%). Regarding AMR determinants, the most significantly (p < 0.05) genes detected by qPCR were sul1 and aadA2 (89% of strains positive), aadA1 and dfrA12 (87.7%), and blaTEM and tet(B) (86.3% and 84.9%, respectively). Up to date information on ST antimicrobial resistance patterns is essential for epidemiological surveillance programs to support animal and public health. The high number of MDR isolates and variability regarding resistance determinants revealed in this study highlights the role of animals reared in extensive systems as a source of resistant Salmonella strains.

Time Series Transcriptomic Analysis of Bronchoalveolar Lavage Cells from Piglets Infected with Virulent or Low-Virulent Porcine Reproductive and Respiratory Syndrome Virus 1.

Porcine reproductive and respiratory syndrome virus (PRRSV) has evolved to escape the immune surveillance for a survival advantage leading to a strong modulation of host's immune responses and favoring secondary bacterial infections. However, limited data are available on how the immunological and transcriptional responses elicited by virulent and low-virulent PRRSV-1 strains are comparable and how they are conserved during the infection. To explore the kinetic transcriptional signature associated with the modulation of host immune response at lung level, a time-series transcriptomic analysis was performed in bronchoalveolar lavage cells upon experimental in vivo infection with two PRRSV-1 strains of different virulence, virulent subtype 3 Lena strain or the low-virulent subtype 1 3249 strain. The time-series analysis revealed overlapping patterns of dysregulated genes enriched in T-cell signaling pathways among both virulent and low-virulent strains, highlighting an upregulation of co-stimulatory and co-inhibitory immune checkpoints that were disclosed as Hub genes. On the other hand, virulent Lena infection induced an early and more marked "negative regulation of immune system process" with an overexpression of co-inhibitory receptors genes related to T-cell and NK cell functions, in association with more severe lung lesion, lung viral load, and BAL cell kinetics. These results underline a complex network of molecular mechanisms governing PRRSV-1 immunopathogenesis at lung level, revealing a pivotal role of co-inhibitory and co-stimulatory immune checkpoints in the pulmonary disease, which may have an impact on T-cell activation and related pathways. These immune checkpoints, together with the regulation of cytokine-signaling pathways, modulated in a virulence-dependent fashion, orchestrate an interplay among pro- and anti-inflammatory responses. IMPORTANCE Porcine reproductive and respiratory syndrome virus (PRRSV) is one of the major threats to swine health and global production, causing substantial economic losses. We explore the mechanisms involved in the modulation of host immune response at lung level performing a time-series transcriptomic analysis upon experimental infection with two PRRSV-1 strains of different virulence. A complex network of molecular mechanisms was revealed to control the immunopathogenesis of PRRSV-1 infection, highlighting an interplay among pro- and anti-inflammatory responses as a potential mechanism to restrict inflammation-induced lung injury. Moreover, a pivotal role of co-inhibitory and co-stimulatory immune checkpoints was evidenced, which may lead to progressive dysfunction of T cells, impairing viral clearance and leading to persistent infection, favoring as well secondary bacterial infections or viral rebound. However, further studies should be conducted to evaluate the functional role of immune checkpoints in advanced stages of PRRSV infection and explore a possible T-cell exhaustion state.

The jigsaw of PRRSV virulence.

Porcine reproductive and respiratory syndrome virus (PRRSV) is the causative agent of the, probably, most economically important disease for the pig industry worldwide. This disease, characterised by producing reproductive failure in sows and respiratory problems in growing pigs, appeared in the late 1980s in the United States and Canada. Since its appearance, strains capable of producing higher mortality rates as well as greater severity in clinical signs and lesions than classical strains have been identified. However, since the first reports of these "virulent" PRRSV outbreaks, no homogeneity and consensus in their description have been established. Moreover, to the authors' knowledge, there is no published information related to the criteria that a PRRSV strain should fulfil to be considered as a "virulent" strain. In this review, we revise the terminology used and gather the information related to the main characteristics and differences in clinical signs, lesions, viral replication and tropism as well as immunological parameters between virulent and classical PRRSV strains and propose a first approximation to the criteria to define a virulent PRRSV strain.

Prevalence of mycoplasma-like lung lesions in pigs from commercial farms from Spain and Portugal.

BACKGROUND: Mycoplasma hyopneumoniae causes a chronic respiratory disease that produces important economic losses due to poor productive performance, increased mortality and costs for several control strategies. The prevalence of mycoplasma-like lesions (MLL) at abattoir has been widely studied in different countries, making use of different scoring systems. However, most of them are difficult to apply in abattoirs with high number of pigs sacrificed per hour. For that reason, it is necessary to adapt the scoring system to the reality of the modern abattoir, even if there is a loss of accuracy. Our purpose was to investigate the prevalence and severity of MLL at abattoirs in Spain and Portugal using a 0 to 5 scoring system adapted to abattoirs with high number of sacrificed pigs per hour and to highlight the histopathological diagnosis as confirmatory method to identify patterns of pneumonia correlated to gross lesions. RESULTS: Cranioventral pulmonary consolidation, a typical MLL, was the most frequent lung lesion (30.97 %) detected at the abattoir, followed by dorsocaudal infarcts with pleurisy (12.51 %) and pleurisy alone (6.26 %). The average score for all examined lungs at abattoir was 1.99 out of 5 points. The histopathological study revealed that the 78.17 % of the randomly selected lungs with MLL presented microscopic lesions compatible with M. hyopneumoniae infection. Most bronchointerstitial and interstitial pneumonia lesions had a chronic course while most suppurative and fibrinous bronchopneumonia lesions had an acute course and a higher degree of severity. The combination of microscopic lesions more frequently observed was bronchointerstitial pneumonia + interstitial pneumonia + suppurative bronchopneumonia. CONCLUSIONS: The prevalence of MLL at abattoir was 30.97 %, however, after microscopic examination the real prevalence of lungs with lesions compatible with M. hyopneumoniae infection was reduced up to 24.21 %. The six more prevalent combinations of lesions in the microscopic study involved the 66.13 % of examined lungs, and in all of them, microscopic lesions characteristic of M. hyopneumoniae infection were found, what supports the importance of M. hyopneumoniae as a primary pathogen in cases of PRDC.

Activation of pro- and anti-inflammatory responses in lung tissue injury during the acute phase of PRRSV-1 infection with the virulent strain Lena.

Porcine reproductive and respiratory syndrome virus (PRRSV) plays a key role in porcine respiratory disease complex modulating the host immune response and favouring secondary bacterial infections. Pulmonary alveolar macrophages (PAMs) are the main cells supporting PRRSV replication, with CD163 as the essential receptor for viral infection. Although interstitial pneumonia is by far the representative lung lesion, suppurative bronchopneumonia is described for PRRSV virulent strains. This research explores the role of several immune markers potentially involved in the regulation of the inflammatory response and sensitisation of lung to secondary bacterial infections by PRRSV-1 strains of different virulence. Conventional pigs were intranasally inoculated with the virulent subtype 3 Lena strain or the low virulent subtype 1 3249 strain and euthanised at 1, 3, 6 and 8 dpi. Lena-infected pigs exhibited more severe clinical signs, macroscopic lung score and viraemia associated with an increase of IL-6 and IFN-γ in sera compared to 3249-infected pigs. Extensive areas of lung consolidation corresponding with suppurative bronchopneumonia were observed in Lena-infected pigs. Lung viral load and PRRSV-N-protein+ cells were always higher in Lena-infected animals. PRRSV-N-protein+ cells were linked to a marked drop of CD163+ macrophages. The number of CD14+ and iNOS+ cells gradually increased along PRRSV-1 infection, being more evident in Lena-infected pigs. The frequency of CD200R1+ and FoxP3+ cells peaked late in both PRRSV-1 strains, with a strong correlation between CD200R1+ cells and lung injury in Lena-infected pigs. These results highlight the role of molecules involved in the earlier and higher extent of lung lesions in piglets infected with the virulent Lena strain, pointing out the activation of routes potentially involved in the restraint of the local inflammatory response.

Perspectives on cervical cancer screening and prevention: challenges faced by providers and patients along the Texas-Mexico border.

BACKGROUND: The Rio Grande Valley (RGV) and Laredo regions located along the Texas-Mexico border consist of seven counties with a population of approximately 1.5 million people and a high uninsured rate (33.5%). Cervical cancer mortality in these border counties is approximately 30% higher than the rest of Texas. The RGV and Laredo areas were studied to better understand the state of access to cervical cancer prevention services along the Texas-Mexico border. METHODS: Data on the population served and the services provided were analyzed to determine the gap between cervical cancer screenings recommended versus those received. Through interviews, we gathered the perspectives of 16 local stakeholders regarding cervical cancer screening for underserved individuals in the region. FINDINGS: It is estimated that 69,139 uninsured women aged 21-64 years in the RGV/Laredo per year are recommended to undergo cervical cancer screening with Papanicolaou (Pap) and/or human papillomavirus (HPV) testing, but only 8941 (12.9%) Pap tests are being performed by the Federally Qualified Health Center (FQHC) serving uninsured women in these regions. Systemic barriers identified include insufficient provider clinical capacity, the high cost of healthcare, and uncertainty about government funding sources. Patient barriers identified include inadequate knowledge on navigating the local healthcare system, low health literacy, lack of money and childcare, an inability to miss work, limited transportation, and fear of deportation. CONCLUSION: Decreasing the disparity between cervical cancer screening services provided and those recommended requires addressing the barriers, identified by local experts, which prevent uninsured women from accessing care. These challenges are being addressed through ongoing programs and collaborations.

Valor pronóstico del antígeno carcinoembrionario hallado en lavados pleurales de pacientes con carcinoma pulmonar / Prognostic value of the carcinoembryonic antigen found in pleural lavage fluid from patients with lung carcinoma

OBJETIVO: Detectar el marcador tumoral antígeno carcino-embrionario (CEA) en lavados pleurales de pacientes con carcinoma pulmonar sin derrame pleural asociado, en el momento de la cirugía, a fin de estudiar sus posibles implicaciones pronósticas. PACIENTES Y MÉTODOS: Se trata de un estudio observacional prospectivo que recoge de forma consecutiva a los pacientes intervenidos de carcinoma pulmonar a los que se realizó lavado pleural de la cavidad torácica al finalizar su apertura (grupo de estudio). Se practicaron la misma técnica y medición en pacientes toracotomizados por enfermedades benignas (grupo control). También se cuantificó el valor del CEA sanguíneo preoperatorio. RESULTADOS: El CEA (mediana) sanguíneo en el grupo de estudio fue de 2,90 ng/ml y en el lavado pleural de 0,40 ng/ml, cifras superiores a las correspondientes del grupo control. Se estableció como punto de corte del CEA en el lavado pleural el valor de 0,30 ng/ml, establecido según la curva de eficacia diagnóstica correspondiente, con una sensibilidad del 68,4% y una especificidad del 35,7%. La gráfica de supervivencia en función de este punto de corte mostró significación estadística (p < 0,05). CONCLUSIONES: Es posible detectar CEA en lavados pleurales de la cavidad torácica en pacientes con carcinoma pulmonar, con valores superiores a los encontrados en pacientes sin enfermedad neoplásica. La medición de dicho valor es un factor pronóstico independiente en la evolución de la enfermedad

OBJECTIVE: To detect the tumor marker carcinoembryonic antigen (CEA) in pleural lavage fluid taken during surgery from patients with pulmonary carcinoma with out associated pleural effusion and assess its possible prognostic implications. PATIENTS AND METHODS: A prospective, observational study was undertaken to include consecutive patients who underwent surgical treatment for lung carcinoma in which pleural lavage was performed prior to closure of the thoracic cavity (study group). The same techniques and measurements were used in patients undergoing thoracotomy for benign disease (control group). The preoperative blood level of CEA was also quantified. RESULTS: In the study group, the median CEA levels in blood and pleural lavage fluid were 2.90 ng/mL and 0.40 ng/mL respectively; these figures are higher than those corresponding to the control group. A CEA level of 0.30 ng/mL in pleural lavage fluid was established as a cutoff point, based on the corresponding receiver operating characteristic curve, with a sensitivity of 68.4% and a specificity of 35.7%. A graph of survival in relation to this cutoff point revealed a statistically significant effect (P<.05). CONCLUSIONS: It is possible to detect CEA in pleural lavage fluid from the thoracic cavity of patients with lung carcinoma. The values obtained are higher than those found in fluid from patients without neoplastic disease, and this parameter functions as an independent predictor of disease course. A substantial number of patients died of causes unrelated to the NSCLC for which they had been treated