Short palate, lung, and nasal epithelial clone 1 (SPLUNC1) level determines steroid-resistant airway inflammation in aging.

Asthma and its heterogeneity change with age. Increased airspace neutrophil numbers contribute to severe steroid-resistant asthma exacerbation in the elderly, which correlates with the changes seen in adults with asthma. However, whether that resembles the same disease mechanism and pathophysiology in aged and adults is poorly understood. Here, we sought to address the underlying molecular mechanism of steroid-resistant airway inflammation development and response to corticosteroid (Dex) therapy in aged mice. To study the changes in inflammatory mechanism, we used a clinically relevant treatment model of house-dust mite (HDM)-induced allergic asthma and investigated lung adaptive immune response in adult (20-22 wk old) and aged (80-82 wk old) mice. Our result indicates an age-dependent increase in airway hyperresponsiveness (AHR), mixed granulomatous airway inflammation comprising eosinophils and neutrophils, and Th1/Th17 immune response with progressive decrease in frequencies and numbers of HDM-bearing dendritic cells (DC) accumulation in the draining lymph node (DLn) of aged mice as compared with adult mice. RNA-Seq experiments of the aged lung revealed short palate, lung, and nasal epithelial clone 1 (SPLUNC1) as one of the steroid-responsive genes, which progressively declined with age and further by HDM-induced inflammation. Moreover, we found increased glycolytic reprogramming, maturation/activation of DCs, the proliferation of OT-II cells, and Th2 cytokine secretion with recombinant SPLUNC1 (rSPLUNC1) treatment. Our results indicate a novel immunomodulatory role of SPLUNC1 regulating metabolic adaptation/maturation of DC. An age-dependent decline in the SPLUNC1 level may be involved in developing steroid-resistant airway inflammation and asthma heterogeneity.

LasR-deficient Pseudomonas aeruginosa variants increase airway epithelial mICAM-1 expression and enhance neutrophilic lung inflammation.

Pseudomonas aeruginosa causes chronic airway infections, a major determinant of lung inflammation and damage in cystic fibrosis (CF). Loss-of-function lasR mutants commonly arise during chronic CF infections, are associated with accelerated lung function decline in CF patients and induce exaggerated neutrophilic inflammation in model systems. In this study, we investigated how lasR mutants modulate airway epithelial membrane bound ICAM-1 (mICAM-1), a surface adhesion molecule, and determined its impact on neutrophilic inflammation in vitro and in vivo. We demonstrated that LasR-deficient strains induce increased mICAM-1 levels in airway epithelial cells compared to wild-type strains, an effect attributable to the loss of mICAM-1 degradation by LasR-regulated proteases and associated with enhanced neutrophil adhesion. In a subacute airway infection model, we also observed that lasR mutant-infected mice displayed greater airway epithelial ICAM-1 expression and increased neutrophilic pulmonary inflammation. Our findings provide new insights into the intricate interplay between lasR mutants, LasR-regulated proteases and airway epithelial ICAM-1 expression, and reveal a new mechanism involved in the exaggerated inflammatory response induced by lasR mutants.

The Role of Lung and Gut Microbiota in the Pathology of Asthma.

Asthma is a common chronic respiratory disease affecting more than 300 million people worldwide. Clinical features of asthma and its immunological and molecular etiology vary significantly among patients. An understanding of the complexities of asthma has evolved to the point where precision medicine approaches, including microbiome analysis, are being increasingly recognized as an important part of disease management. Lung and gut microbiota play several important roles in the development, regulation, and maintenance of healthy immune responses. Dysbiosis and subsequent dysregulation of microbiota-related immunological processes affect the onset of the disease, its clinical characteristics, and responses to treatment. Bacteria and viruses are the most extensively studied microorganisms relating to asthma pathogenesis, but other microbes, including fungi and even archaea, can potently influence airway inflammation. This review focuses on recently discovered connections between lung and gut microbiota, including bacteria, fungi, viruses, and archaea, and their influence on asthma.

Bovine besnoitiosis in an endemically infected dairy cattle herd in Italy: serological and clinical observations, risk factors, and effects on reproductive and productive performances.

Bovine besnoitiosis (Besnoitia besnoiti) is an emerging parasitic disease of cattle in Europe. This study reports a case of bovine besnoitiosis in a dairy farm housing 217 cattle in Italy. A serological screening was performed on the whole herd using the recommended approach of ELISA and confirmatory Western Blot. Seropositive animals were clinically examined to reveal symptoms and lesions of besnoitiosis. Risk factors and the effects of the parasite infection on reproductive and productive performances were evaluated. Histopathology and molecular analyses on tissues from a slaughtered cow affected by the chronic phase of the disease were carried out. An overall seroprevalence of 23.5%, which increased up to 43.5% considering only cows, was recorded. Clinical examination of 33 of the seropositive cows evidenced the presence of tissue cysts in at least one of the typical localizations (sclera, vulva, or skin) in 25 animals. Statistical analysis did not evidence any significative impact of the parasite infection on herd efficiency; however, a decrease of productive parameters was recorded in cows showing cutaneous cysts. Concerning the chronically affected cow, histopathology revealed B. besnoiti tissue cysts in the skin of the neck, rump, hind legs, eyelid and vulva, in the muzzle, in mucosal membranes of the upper respiratory tract, and in the lungs. Parasite DNA was detected also in masseter muscles, tonsils, mediastinal lymph nodes, liver, cardiac muscle, aorta wall, ovaries, uterus, and vulva. Bovine besnoitiosis continues to spread in the Italian cattle population. Breeders and veterinarians should be aware of this parasitic disease, and control programs should be developed based on surveillance through a diagnostic procedure including both clinical examination and laboratory tests.

First morphological and molecular analysis of Eucoleus boehmi like eggs in dogs from Argentina.

The canid parasites Eucoleus aerophilus (syn. Capillaria aerophila) and Eucoleus boehmi (syn. Capillaria boehmi) parasitize the lower and the upper respiratory tract, respectively. Reports and descriptions of these nematodes are scarce in Argentina, possibly due to misdiagnosis of morphologically similar trichuroids eggs, and the lack of knowledge about the species of Eucoleus in this geographical area. Scanning electron microscopy is a useful tool for identification of E. boehmi eggs based on the characteristics of the shell structure which differentiate between species. Molecular analysis complements morphological identification. Until now, there are no studies based on the analysis of E. boehmi eggs in Argentina. The aim of the present work was to study by morphological, morphometric, and molecular analysis, eggs attributable to E. boehmi isolated from dogs naturally infected in Mar del Plata city, Argentina. Eggs isolated from two dog fecal samples were analyzed by light and scanning electron microscopy. A fragment of mitochondrial DNA (mtDNA) of the cytochrome c oxidase subunit I gene (cox1) from eggs was sequenced, and phylogenetic analysis was performed in this study. According to morphological results based on the wall surface ultrastructure, the eggs studied were assigned to E. boehmi. Molecular analysis supported the morphological identification. The divergence of 9-12% with the European isolated could suggest a new geographical genetic variation of E. boehmi, but also question the possible existence of cryptic species. This is the first characterization of E. boehmi eggs in dogs from Argentina.

Taxonomic status of Syngamus nematodes parasitizing passeriform hosts from Central Europe: Morphological, morphometric and molecular identification.

The systematic position and validity of species within genus Syngamus have always been controversial. In this present work, we evaluated the taxonomic status and phylogenetic relationships between three species of Syngamus nematodes (Syngamus trachea, Syngamus taiga and Syngamus merulae) and one taxa, determined only to the generic level, collected from respiratory tracts of passeriform hosts from Central Europe using newly obtained sequences of 2 nuclear markers (internal transcribed spacers 1 and 2 [ITS1, ITS2]) and a fragment of mitochondrial cytochrome oxidase I [COI] gene. Our results clearly showed that S. trachea, S. taiga and S. merulae are valid, molecularly and morphologically distinct species. Moreover, molecular analysis of adult female of Syngamus sp. collected from tracheae of the European robin Erithacus rubecula clearly indicate that these comprise separate species. In the derived phylogeny, the Syngamus clade is divided into two sub-clades: one comprised Syngamus species with a characteristic, well-developed cuticular collar around the oral opening (S. trachea and S. taiga) and a second that groups taxa without or with rudimentary collar (S. merulae and currently sequenced Syngamus sp. from Erithacus rubecula). These results clearly suggest that the degree of collar development (well-developed vs. rudimentary/absent) may be an important phylogenetic feature for determining the structure of the genus Syngamus on subgeneric level. Additionally, our results support historical division of the genus Syngamus into two subgenera S. (Syngamus) and S. (Ornithogamus).

Aspiration of an interesting foreign body: Myiasis.

Myiasis is a rare condition caused by the invasion of tissues by the larvae of flies. Many cases of myiasis involving various human organs have been reported. Tracheopulmonary or intratracheal myiasis is a very unusual and aberrant form of the disease in humans. We present a case of respiratory myiasis after aspiration of larvae by a healthy 8-month-old girl, which cannot be found in the English literature.

Host protective roles of type 2 immunity: parasite killing and tissue repair, flip sides of the same coin.

Metazoan parasites typically induce a type 2 immune response, characterized by T helper 2 (Th2) cells that produce the cytokines IL-4, IL-5 and IL-13 among others. The type 2 response is host protective, reducing the number of parasites either through direct killing in the tissues, or expulsion from the intestine. Type 2 immunity also protects the host against damage mediated by these large extracellular parasites as they migrate through the body. At the center of both the innate and adaptive type 2 immune response, is the IL-4Rα that mediates many of the key effector functions. Here we highlight the striking overlap between the molecules, cells and pathways that mediate both parasite control and tissue repair. We have proposed that adaptive Th2 immunity evolved out of our innate repair pathways to mediate both accelerated repair and parasite control in the face of continual assault from multicellular pathogens. Type 2 cytokines are involved in many aspects of mammalian physiology independent of helminth infection. Therefore understanding the evolutionary relationship between helminth killing and tissue repair should provide new insight into immune mechanisms of tissue protection in the face of physical injury.

Regulation of macrophage and dendritic cell function by pathogens and through immunomodulation in the avian mucosa.

Macrophages (MPh) and dendritic cells (DC) are members of the mononuclear phagocyte system. In chickens, markers to distinguish MPh from DC are lacking, but whether MPh and DC can be distinguished in humans and mice is under debate, despite the availability of numerous markers. Mucosal MPh and DC are strategically located to ingest foreign antigens, suggesting they can rapidly respond to invading pathogens. This review addresses our current understanding of DC and MPh function, the receptors expressed by MPh and DC involved in pathogen recognition, and the responses of DC and MPh against respiratory and intestinal pathogens in the chicken. Furthermore, potential opportunities are described to modulate MPh and DC responses to enhance disease resistance, highlighting modulation through nutraceuticals and vaccination.

New host records of Brazilian pentastomid species.

Pentastomida is a taxon of parasitic organisms infecting generally the respiratory system of vertebrates, mainly reptiles. Although this taxon is very important for understanding the phylogeny of the Metazoa, it has received little attention. In Brazil, there are few collections that include species of pentastomids, that is, only the Helminthological Collection of the Oswaldo Cruz Institute (CHIOC), the Invertebrate Collection of the Zoology Laboratory of the Regional University of Cariri (LAZ-URCA) and the Helminthological Collection of the Botucatu Institute of Biosciences (CHIBB). The present study describes the species of pentastomids deposited in CHIBB. When found, the pentastomids were mounted on slides in Hoyer's medium and identified. Four species of pentastomids were recognised and three others were identified at the genus level, while four new hosts for the species of pentastomids were recorded.

New host records of Brazilian pentastomid species / Novos registros de hospedeiros para espécies de pentastomídeos do Brasil

Pentastomida is a taxon of parasitic organisms infecting generally the respiratory system of vertebrates, mainly reptiles. Although this taxon is very important for understanding the phylogeny of the Metazoa, it has received little attention. In Brazil, there are few collections that include species of pentastomids, that is, only the Helminthological Collection of the Oswaldo Cruz Institute (CHIOC), the Invertebrate Collection of the Zoology Laboratory of the Regional University of Cariri (LAZ-URCA) and the Helminthological Collection of the Botucatu Institute of Biosciences (CHIBB). The present study describes the species of pentastomids deposited in CHIBB. When found, the pentastomids were mounted on slides in Hoyer's medium and identified. Four species of pentastomids were recognised and three others were identified at the genus level, while four new hosts for the species of pentastomids were recorded.

Pentastomida é um táxon de organismos parasitas obrigatórios de sistema respiratório de vertebrados, principalmente répteis. Embora esse táxon seja muito importante para a compreensão da filogenia dos Metazoa, tem recebido pouca atenção. No Brasil, existem poucas coleções que abrigam espécies de pentastomídeos, quais sejam: a Coleção Helmintológica do Instituto Oswaldo Cruz (CHIOC), a Coleção de Invertebrados do Laboratório de Zoologia da Universidade Regional do Cariri (LAZ-URCA) e a Coleção Helmintológica do Laboratório de Parasitologia de Animais Silvestres (LAPAS). O presente trabalho descreve as espécies de pentastomídeos depositados na Coleção Helmintológia do LAPAS. O trato respiratório e as cavidades do corpo dos répteis foram removidos e analisados sob Microscópio Esteroscópico; quando encontrados os pentastomídeos, foram montados slides em meio Hoyer e identificados. Foram identificadas quatro espécies e outras três ficaram identificadas no nível de gênero, tendo sido registrados quatro novos hospedeiros para as espécies de pentastomídeos.

Network model of immune responses reveals key effectors to single and co-infection dynamics by a respiratory bacterium and a gastrointestinal helminth.

Co-infections alter the host immune response but how the systemic and local processes at the site of infection interact is still unclear. The majority of studies on co-infections concentrate on one of the infecting species, an immune function or group of cells and often focus on the initial phase of the infection. Here, we used a combination of experiments and mathematical modelling to investigate the network of immune responses against single and co-infections with the respiratory bacterium Bordetella bronchiseptica and the gastrointestinal helminth Trichostrongylus retortaeformis. Our goal was to identify representative mediators and functions that could capture the essence of the host immune response as a whole, and to assess how their relative contribution dynamically changed over time and between single and co-infected individuals. Network-based discrete dynamic models of single infections were built using current knowledge of bacterial and helminth immunology; the two single infection models were combined into a co-infection model that was then verified by our empirical findings. Simulations showed that a T helper cell mediated antibody and neutrophil response led to phagocytosis and clearance of B. bronchiseptica from the lungs. This was consistent in single and co-infection with no significant delay induced by the helminth. In contrast, T. retortaeformis intensity decreased faster when co-infected with the bacterium. Simulations suggested that the robust recruitment of neutrophils in the co-infection, added to the activation of IgG and eosinophil driven reduction of larvae, which also played an important role in single infection, contributed to this fast clearance. Perturbation analysis of the models, through the knockout of individual nodes (immune cells), identified the cells critical to parasite persistence and clearance both in single and co-infections. Our integrated approach captured the within-host immuno-dynamics of bacteria-helminth infection and identified key components that can be crucial for explaining individual variability between single and co-infections in natural populations.

Corrected misidentification of first instars of the bot flies Cephenemyia apicata and C. jellisoni, with a review of the biology and life history of both species.

First instars obtained from the uteri of host-seeking females, and compared with previously published descriptions of larvae collected from Columbian black-tailed deer (Odocoileus hemionus columbianus), revealed that larvae described as Cephenemyia apicata Bennett & Sabrosky (Diptera: Oestridae) actually are C. jellisoni Townsend. Several new characters are added to supplement the original descriptions of the first instars of these species. Collections of all larvae from deer autopsied during different months established that the first instars of C. apicata undergo a long obligate period of development and growth in the bronchi before molting and migrating to the retropharyngeal pouches of deer. This is the first study to document an obligatory lung phase in the life history of a species of Cephenemyia. The lung phase of C. apicata is compared with that described for Pharyngomyia picta (Meigen) (Diptera: Oestridae) in European red deer. A literature review found that for all Cephenemyia species studied, as well as for P. picta, a 6-8 month period of development as first instars during the fall/winter months ensures that pupal development and eclosion of adults will occur during periods of favourable external environmental conditions.

Isolation of Staphylococcus epidermidis from inflamed upper respiratory tract of an orange-spined hairy dwarf porcupine (Sphiggurus villosus)

The orange-spined hairy dwarf porcupine (Sphiggurus villosus) is a rodent species common in most parts of South America, and little is known about the pathologies that can afflict it. A specimen was delivered at the Wildlife Research and Medical Center (CEMPAS), School of Veterinary Medicine and Animal Husbandry, UNESP, Botucatu, SP, Brazil. The animal showed intense apathy, with purulent secretion in the nasal cavity and fracture of the lumbar spine. Due to the unfavorable prognosis, the porcupine was euthanized and microbiological culture of nasal discharge showed Staphylococcus epidermidis. The antimicrobial resistance test revealed sensitivity to all tested antimicrobials (ampicillin, oxacillin, tetracycline, penicillin G, neomycin, cephalexin, gentamicin, enrofloxacin, ciprofloxacin, cotrimoxazol, cefoxitin and cephalothin). This bacterium is part of the nasal flora of humans and other animals, and may cause infection under certain conditions. In the present study, the infection and colonization by S. epidermidis was the probable cause of the inflammatory process. The sensitivity to all tested antimicrobials suggests that this strain has not been previously exposed to such drugs.

Helminth infestation in birds of prey (Accipitriformes and Falconiformes) in Southern Italy.

Helminth infestation was identified at post mortem examination in 110/116 (95%) raptors belonging to six species in Southern Italy. Pathological changes associated with helminths were observed in 81/110 (74%) of birds. Lesions in the respiratory system were associated with the nematode Serratospiculum tendo only in Falco peregrinus. Lesions in the digestive tract in a range of species of raptors were associated with nematodes (Cheilospirura falconis, Dispharynx falconis, Dispharynx mathewossianae, Physaloptera spp., Procyrnea spp., Procyrnea leptoptera, Synhimantus spp., Synhimantus laticeps, Eucoleus dispar, Porrocaecum spp. and Porrocaecum angusticolle), acanthocephalans (Centrorhynchus buteonis and Centrorhynchus globocaudatus), digeneans (Neodiplostomum spp., Neodiplostomum perlatum, Parastrigea intermedia and Strigea falconis) and a single cestode (Cladotaenia spp.).

Helminth parasites of the wolf Canis lupus from Latvia.

Thirty-four wolves were collected between 2003 and 2008 from throughout Latvia and examined for helminths. A total of 17 helminth species were recorded: the trematode Alaria alata (85.3%); the cestodes Diphyllobothrium latum (2.9%), Echinococcus granulosus (2.9%), Echinococcus multilocularis (5.9%), Mesocestoides lineatus (5.9%), Taenia crassiceps (8.8%), Taenia hydatigena (41.2%), Taenia (ovis) krabbei (8.8%), Taenia multiceps (47.1%), Taenia pisiformis (20.6%), Taenia polyacantha (11.8%), Taenia spp. (8.8%); and the nematodes Ancylostoma caninum (2.9%), Crenosoma vulpis (9.1%), Eucoleus aerophilus (36.4%), Pearsonema plica (41.4%), Trichinella spp. (69.7%), Toxocara canis (5.8%), and Uncinaria stenocephala (41.2%). Alaria alata presented the highest mean intensity (403.8). All animals were infected with at least one species of parasite, while the maximum recorded in one specimen was eight. No differences in the intensity or prevalence of any helminth species were found among the host based on age and gender, except for T. multiceps which was more prevalent in adults than in juveniles.

NMDA receptor-mediated processes in the Parabrachial/Kölliker fuse complex influence respiratory responses directly and indirectly via changes in cortical activation state.

We tested the hypothesis that glutamate, acting via NMDA-type receptors (NMDAr) in the Parabrachial/Kölliker fuse (PBrKF) nucleus of the pons, is involved both directly and indirectly (via changes in cortical activation state) in modulating breathing and ventilatory responses to hypoxia. To this end we examined the effects of MK-801, injected either systemically or directly into the PBrKF, on the breathing patterns of urethane-anaesthetized rats breathing air or an hypoxic gas mixture as electroencephalographic (EEG) activity alternated between State I (awake-like) and State III (NREM sleep-like) EEG patterns. Regardless of EEG state, systemic MK-801 reduced ventilation primarily by reducing tidal volume while microinjection of MK-801 into the PBrKF reduced ventilation by reducing breathing frequency. With both injections, EEG pattern changed from State I to III mimicking the change from wakefulness to NREM sleep that occurs in unanaesthetized rats given MK-801 systemically. Systemic injection of MK-801 delayed and reduced the response to hypoxia while microinjection of MK-801 into the PBrKF did not reduce the HVR but sustained the hypoxic increase in tidal volume well into the post-hypoxic recovery period. Thus, while NMDAr in the PBrKF complex of the pons play a role in modulating sleep/wake-like states as well as changes in breathing pattern associated with changes in cortical activation state, they are neither involved in the hypoxic ventilatory response nor in the change in hypoxic sensitivity associated with the changes in cortical activation state.