Upper Respiratory Disease Research Articles

The pathogenicity of avian metapneumovirus subtype C wild bird isolates in domestic turkeys

BackgroundAvian metapneumovirus subtype C (aMPV/C) causes severe upper respiratory disease in turkeys. Previous report revealed the presence of aMPV/C in wild birds in the southeast regions of the U.S.MethodsIn this study, aMPV/C positive oral swabs from American coots (AC) and Canada geese (CG) were passaged three times in the respiratory tract of specific pathogen free (SPF) turkeys and used as aMPV/C P3 virus isolates in subsequent studies.ResultsWild bird P3 isolates showed similar growth characteristics when compared to virulent aMPV/C in chicken embryo fibroblast ( CEF) cell cultures and their glycoprotein G gene sequence was closely related to the G gene of aMPV/C Colorado reference virus. Three-day-old commercial or SPF turkeys were inoculated oculonasally with wild bird aMPV/C P3 isolates. At 5 and 7 days post-inoculation (DPI), severe clinical signs were observed in both of the AC and CG virus-exposed groups. Viral RNA was detected in tracheal swabs by reverse transcriptase polymerase chain reaction (RT-PCR). In addition, immunohistochemistry showed virus replication in the nasal turbinate and trachea. All virus-exposed turkeys developed positive antibody response by 14 DPI.ConclusionsOur data demonstrate that aMPV/C wild bird isolates induced typical aMPV/C disease in the domestic turkeys.

Beyin biyopsisi akut dissemine ensefalomiyelit tanısı için gerekli midir

Acute disseminated encephalomyelitis (ADEM) is an immunemediated inflammatory demyelinating disorder of the central nervous system. The white matter of the brain and spinal cord is predominantly affected although cortical gray matter, thalamus, and basal ganglia are often involved. A 24-year-old female presented with complaints of headache, instability and with a sixth cranial nerve paresis following an upper respiratory disease. Multiple ringlike contrast enhanced lesions were seen on magnetic resonance imaging (MRI) and the diagnosis was confirmed by a brain biopsy as ADEM. The patient’s complaints ceased and MRI lesions regressed after 1000mg methylprednisolone administration. Brain biopsy is rarely needed to diagnose ADEM and to exclude other infectious or malign processes. An experienced radiologist can usually make a diagnosis of ADEM correctly and during the follow-up period after medical treatment the resolution of MRI abnormalities in serial MRI scanning can also play a supportive role in diagnosing ADEM.

A clonal outbreak of upper respiratory disease in horses caused by Streptococcus equi subsp. zooepidemicus

reduces symptoms and limits virus propagation, but doesn’t completely protect against new infection. Therefore, equine influenza virus (EIV) surveillance is essential as an early warning system to veterinary practitioners, professionals within the equine industry and the concerned institutions. RESPE (French epidemiological network for equine diseases) is the first European network for epidemiosurveillance of major equine diseases based around sentry veterinarians in France. The aim of this present studywas to demonstrate how the RESPE had contributed to a more efficient surveillance of EI in France since 2006. 920 nasopharyngeal swabs sent by RESPE-associated veterinarians were analysed. 582 respiratory samples were sent by non RESPE-associated veterinarians. RNA was extracted from 140 mL of respiratory fluids with the QIAamp Viral RNA minikit. Detectionwas performed by rRT-PCR amplification targeting theM1gene. Phylogenetic analysiswasperformed after a RT-PCR amplification targeting the H1 fragment of hemagglutinin (H3) gene, directly on RNA extracted from nasal swab. Among 920 samples sent by RESPE-associated veterinarians, 121 (13.1%) were positive for EIV divided into 42 premises. Horses infected by EIV were mainly those participating toequestrianmanifestationswithamajorityof French trotters (44.6%), French saddle ponies (23.1%) and ponies (9.9%). Among the 582 samples received from non RESPE-associated veterinarians, 26 (5.1%) EIV positive horseswere detected. The repartition of EIV positive cases is clearly related to the localization of the RESPE veterinarians. The last important outbreak was observed between February and May 2009. 70 horses were found positive by rRT-PCR. Fifteen of the 23 premises, including the Grosbois training yard (index premise), were managed by RESPEassociated veterinarians. The HA1 nucleotide sequence was completely determined on 39 strains and partially on 8 others. All strains analyzed in this study belonged to the American lineage, Floridian sublineage, Clade 1 and Clade 2. Clade 1was identified only during theGrosbois episode. The findings of this study demonstrate that the analysis realized by the RESPE allowed detecting more EIV in France. RESPE also allow characterization of the strains and participate to epidemiological and vaccine efficacy survey. Implementation of this type of network in other countries should reduce the economic losses associated with outbreaks of EI.

Optimization of an in vitro assay to detect Streptococcus equi subsp. equi

Streptococcus equi is the etiologic agent of a highly infectious upper respiratory disease of horses known as strangles. Bacterial culture methods and polymerase chain reaction (PCR) of nasopharyngeal washes and guttural pouch lavages are used routinely to test clinical and carrier animals for the presence of S. equi but no definitive or gold standard test method has been shown to be optimal. We hypothesized that (i) a flocked swab submerged in ten-fold serial dilution suspensions of S. equi prepared in 0.9% NaCl would detect more colony forming units (CFU) than a rayon swab when used to inoculate a blood agar plate, (ii) centrifugation of a 1ml aliquot of each suspension would improve the limit of detection (LOD) by bacterial culture and PCR compared to the culture or PCR of submerged swab samples, (iii) PCR of the centrifuged samples from each suspension would be more sensitive than aerobic culture alone, and (iv) PCR of a 1ml aliquot directly from a sample would be more sensitive than PCR of a sample following submersion of a flocked swab in 1ml saline. Using 7 ten-fold serial dilutions of S. equi in 0.9% NaCl, the LOD for 4 bacterial culture methods and 3 PCR methods were compared. The LOD of direct PCR and flocked swab culture was determined at 1cfu/ml. All PCR methods were equivalent to each other and were more sensitive than any of the culture methods at the lower dilutions. At higher cell densities (>100cfu/ml) flocked swab culture was not statistically better than rayon swab culture, but it was superior to all other methods tested.

Sta c 3 epitopes and their application as biomarkers to detect specific IgE

Sta c 3 was determined to be one of the major allergens from Stachybotrys chartarum sensu lato. This is among the most common fungi on wet building materials. Various cognizant authorities have found that mold and dampness are associated with exacerbation of asthma and increased upper respiratory disease. The primary amino-acid sequence of this allergen was reported, however the IgE-binding epitopes were unknown. Using SPOT-synthesis techniques (SPOTs), one main linear epitope located between V91 and G105 was discovered by Western blot with atopic human sera. This was confirmed by inhibition ELISA assays with both synthesized epitopes and natural Sta c 3. Alanine scanning also revealed R100 and K101 are the critical amino acids for IgE binding. Using the binding-enhanced and alanine substituted epitopes as potential biomarkers, a reverse ELISA method was developed to detect the Sta c 3 sIgE in atopic human sera. This method employed streptavidin-biotinylated mutated epitopes as the capture matrix, probing the possibility of using small peptides for allergen/species specific IgE assays.

PERSISTENT UPPER RESPIRATORY DISEASES IN CHILDREN: ROLE OF GASTROESOPHAGEAL REFLUX STUDIED USING INTRALUMINAL IMPEDANCE AND PH

BackgroundA variety of signs and symptoms of otolaryngological complaints have been attributed to gastroesophageal reflux disease (GERD). However, proof of such an association is spare. Recent studies suggest that in some cases GERD may be connected with upper respiratory diseases in children, and this paper explores the connection. The aim of the study was to estimate the incidence of GERD among children with persistent upper respiratory disease by inserting an electrode into the esophagus. Reflux can be detected as changes in intraluminal resistance brought on by liquid inside the esophagus, and pH measurements can be used to classify the reflux as acidic or nonacidic.Material and MethodsThis was a multicenter, prospective study of children (4–11.5 years old) with persistent upper respiratory disease. Chronic diseases such as allergy, asthma, cystic fibrosis, or congenital anomalies were excluded. The diagnosis of chronic upper tract disease was made by a single laryngologist. All children underwent a 24-hour multichannel intraluminal impedance and pH (MII–pH) study. The diagnosis of GERD was made with BioVIEW analysis software after manual review by a single investigator.ResultsExactly 31 consecutive patients (17 boys, 14 girls, mean age 6.8 years) were enrolled in the study. A diagnosis of GERD was made in 11/31 (35%) patients. A total of 1172 reflux episodes were detected by MII–pH, and 759 (65%) of them were acid, 413 (35%) were nonacid. The most common symptoms were mouth breathing (28/31), snoring (22/31), fetor ex ore (13/31), hawking (4/11), and apnea (2/11). None of the patients declared gastroenterological symptoms of GERD.ConclusionsThe incidence of GERD in children with persistent upper respiratory diseases was high (35%) in our group, but not as high as has been previously assessed. More than 1/3 of reflux episodes were non-acid. Reflux should be taken into consideration as a probable etiology of persistent upper respiratory diseases in children. 24-h pH and impedance monitoring is a more precise way of diagnosing GERD than classic pH-metry

English

The authors mentioned an epizootic  infection for the chronic upper respiratory diseases caused by béta – haemolytic Streptoccocus equi which have affected horses in two localities in Dakar and Rufisque areas (Dakar’s region) in Senegal, during rainy season in 2009. These studies has been done according to clinical aspects and bacteriological and hematological analysis of the samples. The disease has been observed at the end of the rainy season and looks like an aigue form which can evolute near guerison or death. That’s the first case observed and mentioned in Senegal.   Keys words: Bacteriology, chronic upper respiratory disease, dakar, dry season, hematology, infection, rainy season, Rufisque, Streptococcus, Senegal.

Inhibitors of SARS-3CLpro: virtual screening, biological evaluation, and molecular dynamics simulation studies.

SARS-CoV from the coronaviridae family has been identified as the etiological agent of Severe Acute Respiratory Syndrome (SARS), a highly contagious upper respiratory disease that reached epidemic status in 2002. SARS-3CL(pro), a cysteine protease indispensible to the viral life cycle, has been identified as one of the key therapeutic targets against SARS. A combined ligand and structure-based virtual screening was carried out against the Asinex Platinum collection. Multiple low micromolar inhibitors of the enzyme were identified through this search, one of which also showed activity against SARS-CoV in a whole cell CPE assay. Furthermore, multinanosecond explicit solvent simulations were carried out using the docking poses of the identified hits to study the overall stability of the binding site interactions as well as identify important changes in the interaction profile that were not apparent from the docking study. Cumulative analysis of the evaluated compounds and the simulation studies led to the identification of certain protein-ligand interaction patterns which would be useful in further structure based design efforts.

Bordetella avium causes induction of apoptosis and nitric oxide synthase in turkey tracheal explant cultures

Bordetellosis is an upper respiratory disease of turkeys caused by Bordetella avium in which the bacteria attach specifically to ciliated respiratory epithelial cells. Little is known about the mechanisms of pathogenesis of this disease, which has a negative impact in the commercial turkey industry. In this study, we produced a novel explant organ culture system that was able to successfully reproduce pathogenesis of B. avium in vitro, using tracheal tissue derived from 26 day-old turkey embryos. Treatment of the explants with whole cells of B. avium virulent strain 197N and culture supernatant, but not lipopolysaccharide (LPS) or tracheal cytotoxin (TCT), specifically induced apoptosis in ciliated cells, as shown by annexin V and TUNEL staining. LPS and TCT are known virulence factors of Bordetella pertussis, the causative agent of whooping cough. Treatment with whole cells of B. avium and LPS specifically induced NO response in ciliated cells, shown by uNOS staining and diaphorase activity. The explant system is being used as a model to elucidate specific molecules responsible for the symptoms of bordetellosis.

Nuclear localization of influenza B polymerase proteins and their binary complexes

The viral RNA-dependent RNA polymerases of influenza A and B are trimeric complexes of PA, PB1, and PB2 subunits that are crucial for both transcription and replication of the viral genome. Unlike the significant progress made recently in understanding nuclear transport and molecular assembly of influenza A polymerase, little is known about the influenza B polymerase, although influenza B viruses cause severe upper respiratory disease in humans. The aim of this study was to characterize nuclear localization of the influenza B RNA polymerase proteins and binary complexes. We demonstrated that each polymerase protein has a nuclear localization function, and among them, the PB2 protein exclusively locates to the nucleus while PA and PB1 proteins are associated with the cytoplasm and the nucleus. Furthermore, we show that pairwise binary complexes are formed among the influenza B subunits (PA–PB1, PA–PB2, and PB1–PB2) and both PB1–PB2 and PA–PB2 complexes are predominantly associated with the nucleus while the PA–PB1 complex exhibits both nuclear and cytoplasmic fluorescence signals. Results of our studies represent the first step toward the understanding of nuclear transport and molecular assembly within the influenza B polymerase complex.

Kidney transplant Medicare payments and length of stay: associations with comorbidities and organ quality

IntroductionWe investigated associations between pre-transplant comorbidities, length of stay (LOS) and Medicare payments for transplant hospitalization.Material and methodsWe examined United States Renal Data System for 24,963 recipients of first deceased-donor kidney transplants in 1995-2002 for whom Medicare was the primary payer for at least a year pre-transplant. Pre-transplant ICD-9-CM codes from claims were classified with the Charlson and Elixhauser algorithms. Regression models for payments and LOS included: 1) baseline recipient, donor and transplant factors from the Organ Procurement and Transplant Network (OPTN), 2) OPTN variables and individual comorbidities and 3) OPTN variables and counts of Charlson or Elixhauser comorbidities.ResultsFactors most strongly associated with LOS were type I diabetes, cold ischemia time > 36 h, expanded criteria donor (ECD) and donation after cardiac death (DCD). Except for ECD, each was associated with increased payments. Upper respiratory disease, liver disease, peptic ulcer disease, diabetes, cancer and other diseases were also associated with increased LOS and payments. Each additional Charlson comorbidity increased LOS by 2.94% and payments by $471 (Elixhauser results: 1.71% for LOS, $277 for payments). Use of ECD or DCD organs were associated with 10-15% higher LOS and 5% increased Medicare payments for DCD.ConclusionsThis methodology could be used to explore if Medicare reimbursement for transplantation of higher-risk recipients and using non-standard organs is financially adequate and to analyze related questions in other healthcare systems.

Etiologic and epidemiologic analysis of bacterial infectious upper respiratory disease in Thoroughbred horses at the Seoul Race Park

Infectious upper respiratory disease (IURD) of Thoroughbred racehorses has been a frequent problem (29.6% of incidence) at the Seoul Race Park (Korea). Risk factors for IURD include the season with a high transfer rate (summer and fall), the stabling period (≤ 3 months), and age (2 to 3 years old), suggesting that the movement and new environment may have depressed the immune system of the horses and decreased their ability to respond properly to pathogens. The bacterial strains (n = 98) isolated from IURD horses included Pseudomonas spp., Escherichia coli, Staphylococcus spp., Streptococcus equi subsp. equi and zooepidemicus.

A needs-assessment and demographic survey of infection-control and disease awareness in western US animal shelters

A cross-sectional needs-assessment survey was used to characterize animal shelters in a 6-state region in the western US and describe infection-control practices and disease awareness. Survey questions focused on shelter demographics, infection-control practices and policies, awareness and concern over infectious and zoonotic diseases, staff and volunteer training relating to infection-control and disease awareness, use of diagnostic tools, and isolation procedures and protocols. Fifty percent of shelters responded to the survey and represented a wide variety of shelter types, sizes and locations. The top-three diseases of concern to shelters were feline upper respiratory disease (FURD), canine parvovirus and ringworm. Concern over these diseases was greater in open-admission shelters (compared to limited admission or no-kill/sanctuary) (OR 3.7, 95% CI 1.1–12.5) and in shelters with a desire for more zoonotic-disease training (OR = 6.1, 95% CI 1.5–24.8) (compared to shelters desiring infectious-disease training, training on cleaning and disinfection or those who have no need for further training). In 45% of responding shelters many to most animals arrive with infectious diseases. Written protocols for preventive medicine exist in 88% of shelters, cleaning and disinfection protocols in 75%, specific disease protocols for outbreak situations in 36% and infection-control manuals in 15%. Veterinarians are in charge of infection-control in 6% of shelters. Approximately 45% of shelters vaccinate dogs and cats for rabies. Infectious-disease training is provided to 30% of staff and 35% of volunteers upon hire. Overall, volunteers received less training in infectious- and zoonotic-disease identification, prevention and control than staff members. Ninety percent of shelters said they would benefit from training in infectious and zoonotic disease. Results from this study can be used to assess and address needs in animal shelters relating to infection-control, infectious and zoonotic-disease awareness and can help guide development of shelter staff and volunteer training.

An investigation on the presence of Chlamydiaceae in Swedish dogs.

BackgroundBacteria belonging to the family Chlamydiaceae cause a broad spectrum of diseases in a wide range of hosts, including man, other mammals, and birds. Upper respiratory and genital diseases are common clinical problems caused by Chlamydiaceae. Very little is known about chlamydial infections in dogs. Few clinical reports on natural disease in dogs describe mainly conjunctival and upper respiratory signs, and the role of Chlamydiaceae in genital disease is unclear. The present study aimed at studying the prevalence of Chlamydiaceae in healthy dogs and in dogs with genital or upper respiratory disease, including conjunctivitis.MethodsA real-time polymerase chain reaction (PCR) for Chlamydiaceae was used to detect any chlamydial species within this family. Swab samples from the conjunctiva and the mucosal membranes of the oropharynx, rectum and genital tract were taken from 79 dogs: 27 clinically healthy dogs, 25 dogs with clinical signs from the genital tract and 28 dogs with conjunctivitis. There were 52 female and 27 male dogs. From 7 of the male dogs, additional semen samples were analysed.ResultsNo Chlamydiaceae were detected from any dog.ConclusionsAlthough the number of dogs that was included is limited, the results suggest that cases of Chlamydiaceae in dogs probably are related to infection from other species, and that dogs in general do not harbour Chlamydiaceae. Bacteria belonging to the family Chlamydiaceae do not seem to be of major importance for genital or ocular disease in Swedish dogs.

Multilocus Sequence Typing Identifies Evidence for Recombination and Two Distinct Lineages of Corynebacterium diphtheriae

We describe the development of a multilocus sequence typing (MLST) scheme for Corynebacterium diphtheriae, the causative agent of the potentially fatal upper respiratory disease diphtheria. Global changes in diphtheria epidemiology are highlighted by the recent epidemic in the former Soviet Union (FSU) and also by the emergence of nontoxigenic strains causing atypical disease. Although numerous techniques have been developed to characterize C. diphtheriae, their use is hindered by limited portability and, in some instances, poor reproducibility. One hundred fifty isolates from 18 countries and encompassing a period of 50 years were analyzed by multilocus sequence typing (MLST). Strain discrimination was in accordance with previous ribotyping data, and clonal complexes associated with disease outbreaks were clearly identified by MLST. The data produced are portable, reproducible, and unambiguous. The MLST scheme described provides a valuable tool for monitoring and characterizing endemic and epidemic C. diphtheriae strains. Furthermore, multilocus sequence analysis of the nucleotide data reveals two distinct lineages within the population of C. diphtheriae examined, one of which is composed exclusively of biotype belfanti isolates and the other of multiple biotypes.

Snots and snuffles: rational approach to chronic feline upper respiratory syndromes.

While the most common cause of chronic upper respiratory disease signs in cats is viral disease, with subsequent, self-perpetuating inflammation, other, more discrete causes need to be ruled out. These include foreign bodies, bacterial or fungal infections, oral-dental diseases and neoplasia. Any factors contributing to alterations in the structure or function of the upper airways, including inflammation of any cause, will compromise normal function and predispose to chronic damage if the cat is unable to resolve the underlying factors. The chronic feline snuffler is a frustrating patient to treat. The longer the course of disease, the more severe the consequences to affected tissues, and the more debilitated the patient becomes. A logical diagnostic plan to differentiate probable etiologies and to rule out non-viral causes results in appropriate therapeutic choices. Even with a viral etiology, therapies to reduce the pathological consequences of infection may modulate and help control the clinical signs. Some novel choices and drug combinations are discussed in this review. Cats of all ages may be affected. Cats with conformational (breed or malformation) characteristics, such as short or convoluted nasal passages or very small nostrils, are predisposed to unresolving inflammation. Fungal disease is more relevant in specific geographic regions, making inclusion of a travel history important in history collection. Older cats are more likely to have neoplasia-induced signs. This review is directed at all veterinarians who see cats with chronic, recurrent upper respiratory disease.

Health and Reproductive Assessment of a Free-Ranging Gopher Tortoise (Gopherus polyphemus) Population Following Translocation

ABSTRACT In 2004, a subset of a translocated population of gopher tortoises (n = 14; Gopherus polyphemus) were captured to determine the overall health of individuals and to investigate the prevalence of selected pathogens. Additionally, ultrasonography and endoscopic visualization of the reproductive tract were compared as methodologies to accurately determine sex and reproductive status. Tortoises were premedicated with butorphanol and induced with propofol. The hematology and biochemical profile values obtained were within the reference ranges previously reported for gopher tortoises. Tortoises were parasitized with Alaeuris spp. of pinworms and trichostrongyles. Ten of the tortoises (71%) were also parasitized by hemogregarines. None of the animals had clinical signs indicative of acute upper respiratory disease (URTD); however, six animals had signs consistent with previous upper respiratory infections. One tortoise had an antibody titer against Mycoplasma agassizii, considered to be positive, while ...

Identification of Adenoviruses in Fecal Specimens from Wild Chimpanzees (Pan trogylodytes schweinfurthii) in Western Tanzania

DNA of two distinctive adenoviruses was detected in wild chimpanzees in western Tanzania that showed clinical signs of acute, upper respiratory disease, notably coughing. The amplified sequences from part of the capsid hexon gene suggests that one virus is a novel adenovirus serotype candidate and the other virus is a species C adenovirus most closely related to recent isolates from captive chimpanzees in the United States, Simian AdV 37 with 86% nucleic acid identity and Simian AdV 40 with 95% nucleic acid identity, respectively. The species C adenovirus sequences suggest possible recombination with a human adenovirus. The source of these viruses and disease association is not known.

Effects of dietary lysine supplementation on upper respiratory and ocular disease and detection of infectious organisms in cats within an animal shelter

To determine within a cat shelter effects of dietary lysine supplementation on nasal and ocular disease and detection of nucleic acids of Chlamydophila felis, feline calicivirus (FCV), and feline herpesvirus (FHV-1). 261 adult cats. Cats were fed a diet containing 1.7% (basal diet; control cats) or 5.7% (supplemented diet; treated cats) lysine for 4 weeks. Plasma concentrations of lysine and arginine were assessed at the beginning (baseline) and end of the study. Three times a week, cats were assigned a clinical score based on evidence of nasal and ocular disease. Conjunctival and oropharyngeal swab specimens were tested for FHV-1, FCV, and C felis nucleic acids once a week. Data were collected from 123, 74, 59, and 47 cats during study weeks 1, 2, 3, and 4, respectively. By study end, plasma lysine concentration in treated cats was greater than that in control cats and had increased from baseline. There was no difference between dietary groups in the proportion of cats developing mild disease. However, more treated cats than control cats developed moderate to severe disease during week 4. During week 2, FHV-1 DNA was detected more commonly in swab specimens from treated versus control cats. Dietary lysine supplementation in the amount used in our study was not a successful means of controlling infectious upper respiratory disease within a cat shelter. Rather, it led to increases in disease severity and the incidence of detection of FHV-1 DNA in oropharyngeal or conjunctival mucosal swab specimens at certain time points.

Ventral Bulla Osteotomy for Treatment of Otitis Media in a Rabbit

This article describes the management of otitis media in a domestic rabbit ( Oryctolagus cuniculus) that presented with a history of chronic upper respiratory disease. Deep nasal culture yielded a pure growth of Bordetella bronchiseptica susceptible to chloramphenicol. To further evaluate recurring clinical signs after treatment with chloramphenicol, skull radiographs were obtained and showed an increased density in the right tympanic bulla. A ventral bulla osteotomy was performed, and the success of treatment was determined by the resolution of respiratory signs and the absence of increased radiographic density in the right tympanic bulla 5 months postoperatively.