Sheep Lung Research Articles

Exploring transcriptomic mechanisms underlying pulmonary adaptation to diverse environments in Indian rams.

The Changthangi sheep thrive at high altitudes in the cold desert regions of Ladakh, India while Muzaffarnagri sheep are well-suited to the low altitude plains of northern India. This study investigates the molecular mechanisms of pulmonary adaptation to diverse environments by analyzing gene expression profiles of lung tissues through RNA sequencing. Four biological replicates of lung tissue from each breed were utilized to generate the transcriptomic data. Differences in gene expression analysis revealed discrete expression profiles in lungs of each breed. In Changthangi sheep, genes related to immune responses, particularly cytokine signaling, were significantly enriched. Pathway analysis highlighted the activation of NF-kB signaling, a key mediator of inflammation and immune response. Additionally, the gene network analysis indicated a strong association between cytokine signaling, hypoxia-inducible factor (HIF) and NF-kB activation, suggesting a coordinated response to hypoxic stress in lungs of Changthangi sheep. In Muzaffarnagri sheep, the gene expression profiles were enriched for pathways related to energy metabolism, homeostasis and lung physiology. Key pathways identified include collagen formation and carbohydrate metabolism, both of which are crucial for maintaining lung function and structural integrity. Gene network analysis further reinforced this by revealing a strong connection between genes associated with lung structure and function. Our findings shed light on the valuable insights into gene expression mechanisms that enable these sheep breeds to adapt to their respective environments and contribute to a better understanding of high altitude adaptation in livestock.

Multi-Channel Optimization Generative Model for Stable Ultra-Sparse-View CT Reconstruction.

Score-based generative model (SGM) has risen to prominence in sparse-view CT reconstruction due to its impressive generation capability. The consistency of data is crucial in guiding the reconstruction process in SGM-based reconstruction methods. However, the existing data consistency policy exhibits certain limitations. Firstly, it employs partial data from the reconstructed image of the iteration process for image updates, which leads to secondary artifacts with compromising image quality. Moreover, the updates to the SGM and data consistency are considered as distinct stages, disregarding their interdependent relationship. Additionally, the reference image used to compute gradients in the reconstruction process is derived from the intermediate result rather than ground truth. Motivated by the fact that a typical SGM yields distinct outcomes with different random noise inputs, we propose a Multi-channel Optimization Generative Model (MOGM) for stable ultra-sparse-view CT reconstruction by integrating a novel data consistency term into the stochastic differential equation model. Notably, the unique aspect of this data consistency component is its exclusive reliance on original data for effectively confining generation outcomes. Furthermore, we pioneer an inference strategy that traces back from the current iteration result to ground truth, enhancing reconstruction stability through foundational theoretical support. We also establish a multi-channel optimization reconstruction framework, where conventional iterative techniques are employed to seek the reconstruction solution. Quantitative and qualitative assessments on 23 views datasets from numerical simulation, clinical cardiac and sheep's lung underscore the superiority of MOGM over alternative methods. Reconstructing from just 10 and 7 views, our method consistently demonstrates exceptional performance.

Characteristics of lung resistance and elastance associated with tracheal stenosis and intrapulmonary airway narrowing in ex vivo sheep lungs

BackgroundUnderstanding the characteristics of pulmonary resistance and elastance in relation to the location of airway narrowing, e.g., tracheal stenosis vs. intrapulmonary airway obstruction, will help us understand lung function characteristics and mechanisms related to different airway diseases.MethodsIn this study, we used ex vivo sheep lungs as a model to measure lung resistance and elastance across a range of transpulmonary pressures (5–30 cmH2O) and ventilation frequencies (0.125–2 Hz). We established two tracheal stenosis models by inserting plastic tubes into the tracheas, representing mild (71.8% lumen area reduction) and severe (92.1%) obstructions. For intrapulmonary airway obstruction, we induced airway narrowing by challenging the lung with acetylcholine (ACh).ResultsWe found a pattern change in the lung resistance and apparent lung elastance as functions of ventilation frequency that depended on the transpulmonary pressure (or lung volume). At a transpulmonary pressure of 10 cmH2O, lung resistance increased with ventilation frequency in severe tracheal stenosis, whereas in ACh-induced airway narrowing the opposite occurred. Furthermore, apparent lung elastance at 10 cmH2O decreased with increasing ventilation frequency in severe tracheal stenosis whereas in ACh-induced airway narrowing the opposite occurred. Flow-volume analysis revealed that the flow amplitude was much sensitive to ventilation frequency in tracheal stenosis than it was in ACh induced airway constriction.ConclusionsResults from this study suggest that lung resistance and apparent elastance measured at 10 cmH2O over the frequency range of 0.125-2 Hz can differentiate tracheal stenosis vs. intrapulmonary airway narrowing in ex vivo sheep lungs.

Impaired DNA Double-Strand Break Repair in Irradiated Sheep Lung Fibroblasts: Late Effects of Previous Irradiation of the Spinal Thecal Sac.

Children with cancer previously treated with radiotherapy face the likelihood of side effects that can be debilitating or fatal. This study aimed to assess the long-term effect of medulloblastoma radiotherapy on the DNA double-strand break (DSB) repair capability of primary fibroblasts derived from lung biopsies of previously irradiated young sheep. This study included biopsies from three control and five irradiated sheep. The treated sheep had previously received spinal radiotherapy at a total dose of 28 Gy, which is equivalent to pediatric medulloblastoma treatment. Lung biopsies were taken 4 years post-irradiation from high-dose (HD, >18 Gy) and low-dose (LD, <2 Gy) regions. Fifteen cell lines were extracted (six control, four LD and five HD). The cells were irradiated, and DNA DSB repair was analyzed by immunofluorescence. Clonogenic, trypan blue and micronuclei assays were performed. Both the HD and LD cell lines had a significantly higher number of residual γH2AX foci 24 h and a significant decrease in pATM activity post-irradiation compared to the control. There was no statistically significant difference in the clonogenic assay, trypan blue and micronuclei results. Our study showed that a previous irradiation can impair the DNA DSB repair mechanism of ovine lung fibroblasts.

Establishment of 3D cell culture systems with decellularized lung-derived extracellular matrix hydrogel scaffold

Decellularized tissue hydrogels, especially that mimic the native tissue, have a high potential for tissue engineering, three-dimensional (3D) cell culture, bioprinting, and therapeutic agent encapsulation due to their excellent biocompatibility and ability to facilitate the growth of cells. It is important to note that the decellularization process significantly affects the structural integrity and properties of the extracellular matrix, which in turn shapes the characteristics of the resulting hydrogels at the macromolecular level. Therefore, our study aims to identify an effective chemical decellularization method for sheep lung tissue, using a mixing/agitation technique with a range of detergents, including commonly [Sodium dodecyl sulfate (SDS), Triton X-100, and 3-((3-cholamidopropyl) dimethylammonio)-1-propanesulfonate] (CHAPS), and rarely used (sodium cholate hydrate, NP-40, and 3-[N,N-Dimethyl(3-myristoylaminopropyl)ammonio]propanesulfonate) (ASB-14). After the effectiveness of the used detergents on decellularization was determined by histological and biochemical methods, lung derived decellularized extracellular matrix was converted into hydrogel. We investigated the interactions between lung cells and decellularized extracellular matrix using proliferation assay, scanning electron microscopy, and immunofluorescence microscopy methods on BEAS-2B cells in air-liquid interface. Notably, this study emphasizes the effectiveness of ASB-14 in the decellularization process, showcasing its crucial role in removing cellular components while preserving vital extracellular matrix biological macromolecules, including glycosaminoglycans, collagen, and elastin. The resulting hydrogels demonstrated favorable mechanical properties and are compatible with both cell-cell and cell-extracellular matrix interactions.

Detection of Aerobic Bacterial Species Causing Pneumonia in Sheep and Goats and their Susceptibility to Antibiotics in Sudan

Aims: This study was conducted in Khartoum and Gezira States slaughterhouses to investigate the bacterial spp. associated with pneumonia in condemned sheep and goat lungs. Study Design: A total of 2085 animals were examined grossly for respiratory lesions (sheep= 1601 and goats= 484). Place and Duration of Study: Khartoum and Gezira States slaughterhouses during different seasons, from 2018-2020 and conducted in Central Veterinary Research Laboratory, Sudan. Methodology: Bacterial spp. were isolated and identified using analytical profile index strips then more characterization done using Polymerase chain reaction for representative 20 selected isolates and finally antimicrobial susceptibility test was performed. Results: Out of 320 pneumonic lung samples, 116 samples showed pure colonies and 41 were mixed culture while 163 revealed no bacterial growth. The main bacteria isolated were Staphylococcus aureus, Escherichia coli, Pasteurella multocida, Mannheimia haemolytica, Streptococcus spp. Corynebacterium spp. Pseudomonas spp. Staph spp. and Bacillus spp. Twenty representative bacterial isolates examined by PCR technique 10 were Staph aureus and 10 were E. coli to confirm identification. Seven of each were found positive. The Gram-positive bacteria resulted in 41 resistant isolates, 21 intermediate and 141 sensitive to different antibiotics. Ten bacterial species were reported resistant for more than two Antibiotics from different classes mainly tetracycline, methicillin, penicillin and erythromycin. The Gram-negative bacteria resulted in 59 were resistant, 13 intermediate and 108 sensitive to antibiotics. Seventeen of them were multidrug resistance to tetracycline, gentamicin, Imipenem and ceftriaxone. Conclusion: It was concluded that bacteria could be the main cause responsible for pneumonia in sheep and goats, some of these bacterial isolates were resistance to several Antibiotic which have an impact on public health.

Thermography and deep learning for detection of ground beef adulteration

The aim of this study was to evaluate the use of thermal imaging as a rapid and non-destructive method for the detection of adulteration in minced beef. The adulterant materials investigated included avian offal (chicken gizzard) and ovine offal (sheep lung), which have been observed to be used as adulterants in the market. The weight ratios (w/w) of these impurities ranged from 0 to 30 % at 5 % intervals along with the pure samples. Since induction of a temperature gradient helps in thermographic diagnosis, heating and cooling pretreatments were utilized and compared. Convolutional neural network models were developed to identify the adulteration levels in ground beef. The models trained using data of both thermal processes (heating and cooling) were able to detect 8 levels of sheep lung adulteration in ground beef with 99 % accuracy. It was also able to detect the adulteration level of chicken gizzards in the warming method with 99 % accuracy and in the cooling method with 84 % accuracy. Since the method was capable of detecting impurities as low as 5 %, it points to a promising technique with high speed and accuracy for the detection of various types of adulteration in minced meat.

Investigation of Mitochondrial Cytb Gene Region of Both Echinococcus granulosus Eggs from Dogs and Cystic Echinococcosis Isolates Obtained from Sheep and Cattle by Molecular Methods.

We aimed to determine the common Echinococcus granulosus genotypes in Ağrı, Türkiye and to obtain information on the transmission of this parasite. Cystic echinococcosis samples from 100 slaughtered cattle and 100 slaughtered sheep and faecal samples from 200 stray dogs were included in 2021. Collected cyst fluid samples and faces were examined microscopically. DNA was isolated from the germinal membrane of the cysts and from the parasite eggs in the stool samples. The mitochondrial cytb gene region of the parasite was amplified by PCR. Genotypes were determined using the Basic Local Alignment Search Tool (BLAST) after sequence analysis of PCR amplicons. The highest percentage of cysts was found in the lungs of sheep and the liver of cattle. In addition, 75% of sheep cysts and 25.6% of cattle cysts were fertile. Taenia spp./Echinococcus spp. eggs were found in 6% of the faeces of 200 dogs examined microscopically. E. granulosus eggs were detected in 4 out of 50 stool samples analysed by PCR. All samples analysed by sequence analysis were identified as E. granulosus s.s. G1 genotype. Sequence comparison revealed revealed one or more-point mutations in different regions of the five samples. E. granulosus s.s. G1 genotype, known as sheep strain, is common in the Ağrı, Türkiye. The controlled slaughter of livestock, especially sheep, and the avoidance of feeding hydatid cyst organs to dogs, together with public education, were necessary to prevent the spread of the disease.

Assessment of health problems of sheep and goats based on ante-mortem and post-mortem inspection at Addis Ababa Abattoir, Ethiopia.

Ethiopia has a rapidly growing small ruminant sector, which faces low productivity due to husbandry practices and poor health condition of the animals. A study was conducted in Ethiopia's largest municipal abattoir with the objective to assess the health problems of sheep and goats presented for slaughter using standard ante-mortem and post-mortem methodology. A cross-sectional study using systematic random sampling was conducted on 384 sheep and 384 goats from January to July 2014. Soiled skin (69.1%), poor body condition (24.3%), and nostril discharge (19.5%) were common among both species at ante-mortem examination. Gross lesions were frequent in livers (39.7%) and lungs (37.2%), while pneumonia (18.1%) and adhesions (13.8%) were frequent in the lungs of sheep and goats, indicating stress-related illness. Parasitic lesions, especially fasciolosis (19.3%) and hydatid cysts (8.1%) were significantly more common in sheep livers (p 0.05). The direct financial loss from lesions in both species was 1,077,015 ETB or 53,851 USD per year, most of which was estimated to occur from carcass bruising. The findings indicate that reducing parasite burden and preventing carcass bruising through improved handling could significantly increase the profitability of the small ruminant meat sector in Ethiopia.

Examination of adenoviruses with molecular and pathological methods in sheep pneumonia cases

Objective. Reveal adenoviruses (AdV) that cause pneumonia in sheep and examine pathologic changes in the pulmonary and mediastinal lymph nodes of naturally infected adenovirus-positive specimens. Material and method. For this purpose, 1459 lungs of sheep slaughtered in a slaughterhouse were macroscopically examined, and pneumonia lesions were detected in 88 (6.03%) of these. The paraffinized tissue sections of these specimens with pneumonia were examined with the immunohistochemical (IHC) and indirect immunofluorescence (IF) methods, whereas their tissue homogenates were examined using the Antigen ELISA and PCR methods for adenovirus positivity. Results. Accordingly, the prevalence of adenoviruses was determined as 19.3% for IHC, 22.7% for IF, 20.5% for ELISA and 13.6% for PCR. Hematoxylin-eosin (HE) staining was performed to examine histopathological changes in the specimens that were naturally infected with adenoviruses. The histopathological examinations of the naturally infected lung specimens revealed mainly interstitial pneumonia, as well as catarrhal and verminous pneumonia findings. Consequently, it was determined that the most effective methods in the detection of adenoviruses in sheep pneumonias were found respectively as IF, ELISA, IHC and PCR. The finding that adenoviruses were observed only in the mediastinal lymph nodes of some specimens in the immunopathological methods suggested that the latency. Conclusions. The presence of adenoviruses in sheep pneumonia cases was determined with the indirect immunofluorescence, antigen ELISA and PCR methods for the first time. The possibility of the latent nature of adenovirus infection in these species was also discussed for the first time.

Effect of nitric oxide supply on the morphofunctional state of the lungs during cardiopulmonary bypass modelling: an experimental study

Perioperative management of cardiac surgery leads to significant morphofunctional impairment of the lungs, cardiopulmonary bypass (CPB) being the principal contributor. The pathophysiological mechanisms associated with cardiopulmonary bypass include ischaemic-reperfusion injury, oxidative stress and systemic inflammation. Nitric oxide is able to limit the associated damage.Aim of the study: to investigate the effect of nitric oxide supply on morphofunctional state of sheep lungs under simulated cardiopulmonary bypass. Methods. 12 sheep of Altai breed were divided into 2 equal groups. Classical techniques of pulmonary ventilation and cardiopulmonary bypass were simulated in the CPB group. The sheep of the CPB + NO group were supplied with 80 ppm nitric oxide (via respiratory circuit or extracorporeal circulation circuit) during mechanical ventilation and cardiopulmonary bypass. The gas supply started immediately after tracheal intubation and continued until the end of the experiment. In both groups, the cardiopulmonary bypass time was 90 min, after which the sheep were switched to spontaneous circulation and observed for 60 min. Subsequently, blood was sampled, and the animals were withdrawn from the experiment with subsequent collection of histologic specimens.Results. Statistically significant intergroup differences in P / F-index level before the end of the experiment were found (p = 0.041). Nitric oxide supply was associated with decreased infiltrate density in the lung parenchyma (p = 0.006) and increased alveolar area (p &lt; 0.001).Conclusion. Supply of NO during modelling of cardiopulmonary bypass in experimental animals improves the morphological and functional state of the lungs by reducing inflammation, vascular changes and damage to the respiratory part of the lungs (structure of alveoli, alveolar passages and pneumocytes). Clinical studies are needed to investigate pulmonoprotective properties of NO in humans.

Expression and Variations in EPO Associated with Oxygen Metabolism in Tibetan Sheep.

After a long period of adaptive evolution, Tibetan sheep have adapted to the plateau environment in terms of genetics, physiology and biochemistry, but the mechanism of hypoxia adaptation has not been fully elucidated, and the functional genes and molecular mechanisms regulating the hypoxia adaptation of Tibetan sheep need to be further studied. In this study, Tibetan sheep were selected as the research object, and the mRNA expression levels of the hypoxa-related gene EPO in heart, lung, kidney, liver, spleen and longissimus dorsi muscle of Hu sheep (100 m) and Tibetan sheep at different altitudes (2500 m, 3500 m, 4500 m) were assessed by RT-qPCR. The SNPs loci were detected by sequencing and Kompetitive Allele-Specific PCR (KASP) technology, then the correlation between genetic polymorphism and blood gas was analyzed. The results show that the expression of the EPO gene was the highest in the kidney, indicating that the expression of EPO gene had tissue differences. The expression levels of the EPO gene in the heart, lung and liver of Tibetan sheep at a 4500 m altitude were significantly higher than those in Hu sheep (p < 0.05), and the levels in the hearts of Tibetan sheep increased with the increase in altitude. Three mutations were identified in the EPO gene, the SNPs (g.855 A > C) in exon 1 and the SNPs (g.1985 T > G and g.2115 G > C) in exon 4, which were named EPO-SNP1, EPO-SNP2 and EPO-SNP3, respectively, and all three SNPs showed three genotypes. Correlation analysis showed that g.2115 G > C sites were significantly correlated with pO2 (p < 0.05), and haplotype combinations were significantly correlated with pO2 (p < 0.05). Thesee results suggest that the expression of the EPO gene is altitude-differentiated and organ-differentiated, and the EPO gene variants have significant effects on pO2, which may be beneficial to the adaptation of Tibetan sheep to hypoxia stress.

Positive end-expiratory pressure and surfactant administration mode influence function in ex-vivo premature sheep lungs.

Respiratory distress syndrome often necessitates endotracheal surfactant administration in extremely preterm infants. Our study aimed to explore a multi-modal simulation tool for investigating treatment strategies in exvivo sheep lungs during spontaneous breathing. An electromechanical lung simulator (xPULM) mimicking spontaneous breathing was coupled with a non-aerated premature sheep lung, replicating a premature respiratory system. Changes in tidal volume for different positive end-expiratory pressure (PEEP) levels prior to and after either bolus or nebulised surfactant administration were compared. In two preterm sheep lungs, we observed a progressive decline in tidal volume with increasing PEEP levels prior to surfactant delivery from 0.30 ± 0.01 mL at zero PEEP to 0.04 ± 0.01 mL at 15 cmH2O PEEP. Our measurements showed that both bolus (p < 0.05) and nebulised (p < 0.05) surfactant administration resulted in a significant increase in tidal volume, with no significant difference (p = 0.71) between the two methods. The experimental setup demonstrated the feasibility of xPULM for investigating the effectiveness of different PEEP levels and modes of surfactant administration with respect to tidal volume in premature sheep lungs. The lack of adequate lung water resorption in our model warrants further investigations.

First Molecular Characterization of Small Ruminant Lentiviruses Detected in Romania

Small ruminant lentiviruses (SRLVs) are a group of retroviruses that cause multisystem chronic diseases in goats and sheep and lead to production losses in these animals, negatively affecting animal health and welfare. Although molecular characterization of SRLV field isolates has been performed in many countries, there is currently no information on SRLV genotypes circulating in sheep and goats in Romania. Therefore, the main objective of this study was to conduct a molecular and phylogenetic analysis of SRLVs from Romania and determine the degree of genetic relatedness of the obtained sequences to other known SRLV reference strains. A total of 81 sheep lung tissue samples and 41 sheep lung lymph node samples were tested using nested real-time PCR, and samples positive for real-time PCR were used to amplify an 800 bp gag-pol fragment and an overlapping 625 bp fragment of the gag gene. Pairwise DNA distance and phylogenetic analysis showed that the Romanian SRLV strains were closely related to the A2 and A3 strains based on gag-pol sequences and to the A3 and A17 subtypes based on gag sequences. No recombination events were found. Our results revealed that the Romanian sequences have similar epitope patterns to other existing subtypes, although E/K and R/K mutations in epitope 3 were found only in the Romanian sequences, which may have potential value in serological diagnosis. This study is the first report on the genetic characterization of SRLV strains circulating in Romania and provides new information on SRLV heterogeneity. Further detailed studies should be conducted to better understand the divergence of SRLV Romanian strains.

Ecological, epizootic, sanitary and hygienic problems of echinococcosis in domestic goats and dogs in the North Caucasus

Echinococcal invasion, included by the World Health Organization in the list of especially socially dangerous parasitic zoonoses, is an unresolved epidemic, epizootic and sanitary-hygienic problem for the population and animals in more than 250 countries of the world. The purpose of the research was to study the epizootic and sanitary-hygienic problems of echinococcosis of domestic goats in the North Caucasus region. In 2019-2022, the research was carried out in 3 climatic zones of Kabardino-Balkaria. The objects were liver and lungs of 120 slaughter goats. The intestines were dissected from 15 dog corpses slaughtered in accordance with the International Rules for the Humane Treatment of Animals (2012). Monitoring of echinococcosis in dogs and goats was performed by the method of autopsy of organs and tissues according to K.I. Scriabin (1928). The number of eggs of taeniid type in faeces of dogs and in 520 samples of water from natural reservoirs of the Terek river basin was determined according to the guidelines of Russian Research Institute of Parasitology (1986). In plain climatic zone, index of occurrence of cystic Echinococcus granulosus invasion in liver and lungs of goats was 18 %, in foothill zone - 24 %, in mountain zone - 28 % (on average, 25 %). Extracted from liver and lungs of sheep, transparent cysts of echinococcus contained protoscolexes in 100 % of the samples, which indicates the likelihood of foci involving dogs and wild predators in the region. Natural reservoirs of Kabardino-Balkaria are also large biotopes of teniosis. Contamination of river water samples with taeniid eggs is 11.037.0 %. The region belongs to the group of subjects with high rates of zoonosis in animals due to violation of the terms of deworming for priotary dogs, lack of sanitary and preventive work with the population. According to the epizootic situation for animal echinococcosis, the Kabardino-Balkarian Republic can be attributed to disadvantaged regions of the Russian Federation, as there is an increase in the number of dogs with echinococcosis and goats infected with fertile cysts. Sanitary and hygienic pollution of the Terek river and its tributaries, Malka, Baksan, Cherek and Chegem, with eggs of cestodes is a threat to the spread of echinococcosis among animals and humans.

Transcriptome analysis reveals molecular regulation mechanism of Tibet sheep tolerance to high altitude oxygen environment

As one of the most important livestock breeds on the Qinghai-Tibet Plateau, Tibetan sheep are of great importance to the local economy, agriculture and culture. Its adaptive mechanism in low temperature and low oxygen at highland altitudes has not been reported. In this study, transcriptome sequencing was used to analyze the heart, liver, spleen, lung, kidney, and muscle tissue of sheep at low and highland altitudes. LOC101112291, SELENOW, COL3A1, GPX1, TMSB4X and HSF4 were selected as candidate genes for adapting to plateau characteristics in Tibet Sheep. Besides, glutathione metabolism, arachidonic acid metabolism, nucleotide excision repair, regulation of actin cytoskeleton, protein digestion and absorption, thyroid hormone synthesis, relaxation signaling pathways may play important roles in the adaptation to plateau hypoxia, and cold tolerance. Structural analysis also showed that sequencing genes related to the adaptation mechanism of Tibet sheep to highland altitude. This study will lay a certain foundation for Tibet sheep research.

Effect of some antiseptics on elastase production from Pseudomonas aeruginosa.

The present study was carried out to investigate the effect of clinically used antiseptics on elastase production from Pseudomonas aeruginosa. 39 clinical isolates were collected from wounds 10 (25.64%) and burns 29 (74.35%) from hospitalized patients in Baghdad city. Elastin preparation by the autoclaving method yielded 10.5 gm of elastin powder from (250 gm) of sheep's lungs, 3.6 gm from (50 gm) of sheep's bladder, and sheep's ligamentum nuchae (65 gm) yielded 15 gm. All P. aeruginosa isolates were tested for their ability to produce elastase by being cultivated on elastin nutrient agar and observed for the enzyme's activity. The bacteria that make elastase grew, and a clear border emerged surrounding the growth after 24 hours. 32 (82.05%) of P. aeruginosa isolates produced the enzyme on the elastin nutrient agar. Elastase-producing P. aeruginosa was tested quantitatively using the ELISA reader and spectrophotometer at (A495) to detect the released amount of Congo red dye from the degradation of the elastin Congo red. P. aeruginosa (P41) showed the highest elastolytic activity; thus, it was selected to determine the effect of the sub-MIC of the antiseptics on elastase production. The results showed that acetic acid was the best agent to inhibit elastase production, followed by silver nitrate, hydrogen peroxide and ethanol in descending order. Keywords: Elastase; Elastin; Pseudolysin; Antiseptics; Acetic acid; Pseudomonas aeroginosa

Characterization of Lung Microbiomes in Pneumonic Hu Sheep Using Culture Technique and 16S rRNA Gene Sequencing.

Hu sheep, a locally bred species in China known for its high productivity, is currently suffering from pneumonia. Here, we combine high-throughput 16SrRNA gene sequencing and bacterial culturing to examine the bacterial community in pneumonic Hu Sheep lungs (p < 0.05). The results showed that the abundance and diversity of lung bacteria in healthy sheep were significantly higher than those in pneumonia sheep (p = 0.139), while there was no significant difference between moderate and severe pneumonia. Furthermore, the composition of the lung microbiota community underwent significant alterations between different levels of pneumonia severity. The application of LEfSe analysis revealed a notable enrichment of Mannheimiae within the lungs of sheep afflicted with moderate pneumonia (p < 0.01), surpassing the levels observed in their healthy counterparts. Additionally, Fusobacterium emerged as the prevailing bacterial group within the lungs of sheep suffering from severe pneumonia. Integrating the results of bacterial isolation and identification, we conclusively determined that Mannheimia haemolytica was the primary pathogenic bacterium within the lungs of sheep afflicted with moderate pneumonia. Furthermore, the exacerbation of pneumonia may be attributed to the synergistic interplay between Fusobacterium spp. and other bacterial species. Our results provide new insights for guiding preventive and therapeutic measures for pneumonia of different severities in sheep.

Pharmacological blockade of the interleukin-1 receptor suppressed Escherichia coli lipopolysaccharide-induced neuroinflammation in preterm fetal sheep

Intraamniotic inflammation is associated with preterm birth, especially in cases occurring before 32 weeks' gestation, and is causally linked with an increased risk for neonatal mortality and morbidity. Targeted anti-inflammatory interventions may assist in improving the outcomes for pregnancies impacted by intrauterine inflammation. Interleukin-1 is a central upstream mediator of inflammation. Accordingly, interleukin-1 is a promising candidate target for intervention therapies and has been targeted previously using the interleukin-1 receptor antagonist, anakinra. Recent studies have shown that the novel, noncompetitive, allosteric interleukin-1 receptor inhibitor, rytvela, partially resolved inflammation associated with preterm birth and fetal injury. In this study, we used a preterm sheep model of chorioamnionitis to investigate the anti-inflammatory efficacy of rytvela and anakinra, administered in the amniotic fluid in the setting of intraamniotic Escherichia coli lipopolysaccharide exposure. We hypothesized that both rytvela and anakinra would reduce lipopolysaccharide-induced intrauterine inflammation and protect the fetal brain. Ewes with a singleton fetus at 105 days of gestation (term is ∼150 days) were randomized to one of the following groups: (1) intraamniotic injections of 2 mL saline at time=0 and time=24 hours as a negative control group (saline group, n=12); (2) intraamniotic injection of 10 mg Escherichia coli lipopolysaccharide in 2 mL saline and intraamniotic injections of 2 mL saline at time=0 hours and time=24 hours as an inflammation positive control group (lipopolysaccharide group, n=11); (3) intraamniotic injection of Escherichia coli lipopolysaccharide in 2 mL saline and intraamniotic injections of 2.5 mg rytvela at time=0 hours and time=24 hours to test the anti-inflammatory efficacy of rytvela (lipopolysaccharide + rytvela group, n=10); or (4) intraamniotic injection of Escherichia coli lipopolysaccharide in 2 mL saline and intraamniotic injections of 100 mg anakinra at time=0 hours and time=24 hours to test the anti-inflammatory efficacy of anakinra (lipopolysaccharide + anakinra group, n=12). Amniotic fluid was sampled at time 0, 24, and 48 hours (ie, at each intervention and at delivery). Fetal umbilical cord blood was collected at delivery for differential blood counts and chemical studies. Inflammation was characterized by the analysis of fetal tissue cytokine and chemokine levels using quantitative polymerase chain reaction, enzyme-linked inmmunosorbent assay, and histology. The primary study outcome of interest was the assessment of anakinra and rytvela brain-protective effects in the setting of Escherichia coli lipopolysaccharide-induced intrauterine inflammation. Secondary outcomes of interest were to assess protection from fetal and intrauterine (ie, amniotic fluid, chorioamnion) inflammation. Intraamniotic administration of lipopolysaccharide caused inflammation of the fetal lung, brain, and chorioamnionitis in preterm fetal sheep. Relative to treatment with saline only in the setting of lipopolysaccharide exposure, intraamniotic administration of both rytvela and anakinra both significantly prevented periventricular white matter injury, microglial activation, and histologic chorioamnionitis. Anakinra showed additional efficacy in inhibiting fetal lung myeloperoxidase activity, but its use was associated with metabolic acidaemia and reduced fetal plasma insulin-like growth factor-1 levels at delivery. Intraamniotic administration of rytvela or anakinra significantly inhibited fetal brain inflammation and chorioamnionitis in preterm fetal sheep exposed to intraamniotic lipopolysaccharide. In addition, anakinra treatment was associated with potential negative impacts on the developing fetus.

Utilization of endogenous albumin trafficking pathways in the lungs has potential to modestly increase the lung interstitial access and absorption of drug delivery systems after inhaled administration

ABSTRACT Objectives Drug delivery systems typically show limited access to the lung interstitium and absorption after pulmonary delivery. The aim of this work was to undertake a proof-of-concept investigation into the potential of employing endogenous albumin and albumin absorption mechanisms in the lungs to improve lung interstitial access and absorption of inhaled drug delivery systems that bind albumin. Methods The permeability of human albumin (HSA) through monolayers of primary human alveolar epithelia, small airway epithelia, and microvascular endothelium were investigated. The pulmonary pharmacokinetics of bovine serum albumin (BSA) was also investigated in efferent caudal mediastinal lymph duct-cannulated sheep after inhaled aerosol administration. Results Membrane permeability coefficient values (Papp) of HSA increased in the order alveolar epithelia<small airway epithelia<microvascular endothelium, where the permeability of HSA through small airway and microvascular endothelia were approximately 4- and 28-fold higher than alveolar epithelia, respectively. Only 6.5% of the delivered BSA aerosol dose was absorbed from the lungs of sheep over 5 days, although half of the absorbed dose was absorbed via the lung lymph. Conclusion Drug delivery systems that bind endogenous albumin may show a modest increase in lung permeability and absorption after inhaled delivery compared to systems that do not efficiently bind albumin.