Human Respiratory Syncytial Virus and Streptococcus pneumoniae Infection in Wild Bonobos.

Despite being important conservation tools, tourism and research may cause transmission of pathogens to wild great apes. Investigating respiratory disease outbreaks in wild bonobos, we identified human respiratory syncytial virus and Streptococcus pneumoniae as causative agents. A One Health approach to disease control should become part of great ape programs.

Evidence for Human Streptococcus pneumoniae in wild and captive chimpanzees: A potential threat to wild populations.

Habituation of wild great apes for tourism and research has had a significant positive effect on the conservation of these species. However, risks associated with such activities have been identified, specifically the transmission of human respiratory viruses to wild great apes, causing high morbidity and, occasionally, mortality. Here, we investigate the source of bacterial-viral co-infections in wild and captive chimpanzee communities in the course of several respiratory disease outbreaks. Molecular analyses showed that human respiratory syncytial viruses (HRSV) and human metapneumoviruses (HMPV) were involved in the etiology of the disease. In addition our analysis provide evidence for coinfection with Streptococcus (S.) pneumoniae. Characterisation of isolates from wild chimpanzees point towards a human origin of these bacteria. Transmission of these bacteria is of concern because - in contrast to HRSV and HMPV - S. pneumoniae can become part of the nasopharyngeal flora, contributing to the severity of respiratory disease progression. Furthermore these bacteria have the potential to spread to other individuals in the community and ultimately into the population. Targeted vaccination programs could be used to vaccinate habituated great apes but also human populations around great ape habitats, bringing health benefits to both humans and wild great apes.

In situ pneumococcal vaccine production and delivery through a hybrid biological-biomaterial vector.

The type and potency of an immune response provoked during vaccination will determine ultimate success in disease prevention. The basis for this response will be the design and implementation of antigen presentation to the immune system. Whereas direct antigen administration will elicit some form of immunological response, a more sophisticated approach would couple the antigen of interest to a vector capable of broad delivery formats and designed for heightened response. New antigens associated with pneumococcal disease virulence were used to test the delivery and adjuvant capabilities of a hybrid biological-biomaterial vector consisting of a bacterial core electrostatically coated with a cationic polymer. The hybrid design provides (i) passive and active targeting of antigen-presenting cells, (ii) natural and multicomponent adjuvant properties, (iii) dual intracellular delivery mechanisms, and (iv) a simple formulation mechanism. In addition, the hybrid format enables device-specific, or in situ, antigen production and consolidation via localization within the bacterial component of the vector. This capability eliminates the need for dedicated antigen production and purification before vaccination efforts while leveraging the aforementioned features of the overall delivery device. We present the first disease-specific utilization of the vector toward pneumococcal disease highlighted by improved immune responses and protective capabilities when tested against traditional vaccine formulations and a range of clinically relevant Streptococcus pneumoniae strains. More broadly, the results point to similar levels of success with other diseases that would benefit from the production, delivery, and efficacy capabilities offered by the hybrid vector.

Pneumococcal colonization and invasive disease studied in a porcine model.

BACKGROUND: Streptococcus pneumoniae, a Gram-positive bacterium carried in the human nasopharynx, is an important human pathogen causing mild diseases such as otitis media and sinusitis as well as severe diseases including pneumonia, meningitis and sepsis. There is a strong resemblance between the anatomy, immunology and physiology of the pig and human species. Furthermore, there are striking similarities between S. suis pathogenesis in piglets and S. pneumoniae pathogenesis in humans. Therefore, we investigated the use of piglets as a model for pneumococcal colonization and invasive disease. RESULTS: Intravenous inoculation of piglets with an invasive pneumococcal isolate led to bacteraemia during 5 days, showing clear bacterial replication in the first two days. Bacteraemia was frequently associated with fever and septic arthritis. Moreover, intranasal inoculation of piglets with a nasopharyngeal isolate led to colonization for at least six consecutive days. CONCLUSIONS: This demonstrates that central aspects of human pneumococcal infections can be modelled in piglets enabling the use of this model for studies on colonization and transmission but also on development of vaccines and host-directed therapies. Moreover this is the first example of an animal model inducing high levels of pneumococcal septic arthritis.

Structure of the pneumococcal l,d-carboxypeptidase DacB and pathophysiological effects of disabled cell wall hydrolases DacA and DacB.

Bacterial cell wall hydrolases are essential for peptidoglycan turnover and crucial to preserve cell shape. The d,d-carboxypeptidase DacA and l,d-carboxypeptidase DacB of Streptococcus pneumoniae function in a sequential manner. Here, we determined the structure of the surface-exposed lipoprotein DacB. The crystal structure of DacB, radically different to that of DacA, contains a mononuclear Zn(2+) catalytic centre located in the middle of a large and fully exposed groove. Two different conformations were found presenting a different arrangement of the active site topology. The critical residues for catalysis and substrate specificity were identified. Loss-of-function of DacA and DacB altered the cell shape and this was consistent with a modified peptidoglycan peptide composition in dac mutants. Contrary, an lgt mutant lacking lipoprotein diacylglyceryl transferase activity required for proper lipoprotein maturation retained l,d-carboxypeptidase activity and showed an intact murein sacculus. In addition we demonstrated pathophysiological effects of disabled DacA or DacB activities. Real-time bioimaging of intranasal infected mice indicated a substantial attenuation of ΔdacB and ΔdacAΔdacB pneumococci, while ΔdacA had no significant effect. In addition, uptake of these mutants by professional phagocytes was enhanced, while the adherence to lung epithelial cells was decreased. Thus, structural and functional studies suggest DacA and DacB as optimal drug targets.

Management of severe respiratory tract disease caused by human respiratory syncytial virus and Streptococcus pneumoniae in captive chimpanzees (Pan troglodytes).

Chimpanzees (Pan troglodytes) are susceptible to many viral and bacterial pathogens of human origin. This case series reports an acute outbreak of respiratory disease due to human respiratory syncytial virus and Streptococcus pneumoniae in a single group of 30 captive chimpanzees. Both pathogens are potentially zoonotic. The diagnosis was made antemortem and enabled a targeted response to the outbreak; but it more importantly, prompted improvements to the disease surveillance, biosecurity for risk mitigation and risk communication protocols within the zoo. A defined zoonotic disease risk communication pathway provides a model for management and compliance requirements for other collections.

Isolation and antibiotic sensitivity of Streptococcus pneumoniae infections with involvement of multiple organs in lambs.

Respiratory diseases particularly lamb pneumonia is a multifactorial disease involving the interaction between host, etiological agent and environment. The present study was carried out to determine the causative agent of an outbreak of pneumonia in a sheep flock and to establish its pathogenicity and public health importance. The incidence occurred in sheep unit at Madhurikund farm of University (DUVASU), Mathura, Uttar Pradesh, India. At the time of incidence, the population of sheep at the farm was 90. Affected animals were clinically examined and nasal swabs and blood samples were collected from live animals, while morbid materials were collected from dead animals after postmortem examination. The etiological agent was isolated and characterized with conventional microbiological and biochemical methods. Streptococcus pneumoniae was the bacteria isolated from blood, different organs and cerebrospinal fluid. The antibiotic sensitivity revealed resistant to multiple drugs viz., penicillin, tetracycline, erythromycin, chloramphenicol, enrofloxacin and ciprofloxacin. Pathological examination revealed multiple involvements of organs with different degrees of inflammation and haemorrhages of the lower respiratory tract, lungs, liver, heart and kidney. Further, its pathogenicity was established by histopathological examination. In conclusion, presence of multi drug resistant Streptococcus pneumoniae in weaning lambs with the involvement of multiple organs appears to be an emerging zoonotic threat to human particularly in shepherds. This seems to be the first report of isolation of multi drug resistant Streptococcus pneumoniae from outbreak in lambs with multiple organ involvement in India.

Susceptibility of coagulase-negative staphylococci to a kanamycin and cefalexin combination.

A combination of kanamycin and cefalexin was licensed in Europe in 2008 to treat bovine clinical mastitis. Preliminary broth and disk clinical breakpoints for this antibiotic combination have been proposed for Staphylococcus aureus, Streptococcus dysgalactiae, Streptococcus uberis, and Escherichia coli. This study indicates that these proposed breakpoints also hold for coagulase-negative staphylococci (CNS), a group of bacteria frequently isolated in milk samples from cows with clinical mastitis. The data show that clinical bovine mastitis isolates of CNS from Europe have a high degree of susceptibility to the kanamycin/cefalexin combination, with minimal resistance to either agent alone. The use of the available kanamycin and cefalexin combination disk for testing the susceptibility of bovine mastitis isolates of Staph. aureus, Strep. uberis, Strep. dysgalactiae, and E. coli is also reliable for use in the testing of CNS, as disk results correlated with broth minimum inhibitory concentrations. The study reports, for the first time, the approved Clinical Laboratory Standards Institute quality control ranges for the kanamycin/cefalexin combination and wild-type cutoff values for major bacterial pathogens implicated in bovine mastitis.

A rhesus macaque model of Streptococcus pneumoniae carriage.

BACKGROUND: Nasopharyngeal colonization by Streptococcus pneumoniae precedes pneumococcal disease. Elucidation of procedures to prevent or eradicate nasopharyngeal carriage in a model akin to the human would help to diminish the incidence of both pneumonia and invasive pneumococcal disease. METHODS: We conducted a survey of the nasopharynx of infant rhesus macaques from our breeding colony, in search of natural carriers of S. pneumoniae. We also attempted experimental induction of colonization, by nasopharyngeal instillation of a human S. pneumoniae strain (19F). RESULTS: None of 158 colony animals surveyed carried S. pneumoniae in the nasopharynx. Colonization was induced in eight of eight infant rhesus by nasopharyngeal instillation and lasted 2weeks in 100% of the animals and 7weeks in more than 60%. CONCLUSION: Rhesus macaques are probably not natural carriers of S. pneumoniae. The high rate and duration of colonization obtained in our experiments indicates that the rhesus macaque will serve as a human-like carriage model.

Isolation, identification of Streptococcus pneumoniae from infected rhesus monkeys and control efficacy.

BACKGROUND: Streptococcus pneumoniae can cause a wide variety of illnesses. Primate animals can be infected by the pneumococcus. A disease occurred among rhesus monkeys in winter 2006. METHODS: Routine clinical observation, necropsies, bacteriological examinations were conducted, and PCR, pathogenicity to BALB/c mice and antibiotic susceptibility test were examined additionally. RESULTS: We conclude that the agent is S. pneumoniae. Based on the antibiotic susceptibility test, a dose of 20 mg/kg body weight daily of Erythromycin was given intramuscular injection for 5 days, resulting in the disappearance of clinical signs, and no newly case reappear be observed till today. CONCLUSIONS: Therefore, it is suggested that the outbreak of respiratory disease in the rhesus monkeys was because of transmission of S. pneumoniae among rhesus monkeys. The antibiotic therapy finding underscores the utility of Erythromycin to cure the infected rhesus monkeys without causing side effects and without contributing to the further development of antibiotic resistance.

New Streptococcus pneumoniae clones in deceased wild chimpanzees.

In wild chimpanzees in the Taï National Park, Côte d'Ivoire, sudden deaths which were preceded by respiratory problems had been observed since 1999. Two new clones of Streptococcus pneumoniae were identified in deceased apes on the basis of multilocus sequence typing analysis and ply, lytA, and pbp2x sequences. The findings suggest that virulent S. pneumoniae occurs in populations of wild chimpanzees with the potential to cause infections similar to those observed in humans.

Streptococcus pneumoniae-associated cellulitis in a two-month-old Domestic Shorthair kitten.

An approximately 2-month-old, reproductively intact female Domestic Shorthair kitten was presented to the Mississippi Veterinary Research and Diagnostic Laboratory with a history of possible trauma to the left shoulder region while playing with children, and was found dead the following day. Marked swelling, with subcutaneous edema and hemorrhages, was observed in the left forelimb. Severe pleocellular, but largely suppurative cellulitis, fasciitis, and interstitial myositis with edema were observed microscopically in sections from the affected limb. Massive numbers of gram-positive diplococci also were observed. Other pathologic changes included moderate interstitial pneumonia, mild cholangitis, lymph node hemorrhage, gastrointestinal nematodiasis, mild enteritis, and mild interstitial nephritis. Bacteriologic culture identified Streptococcus pneumoniae as the causative agent, which was confirmed by polymerase chain reaction amplification of the pneumolysin gene from chromosomal DNA of the isolate.

Streptococcal meningoencephalitis in a dog.

A 5.5-year-old French bulldog was presented with acute neck pain and a short history of central vestibular syndrome. A marked neutrophilic pleocytosis and numerous gram-positive cocci were evident on cerebrospinal fluid (CSF) cytology. Streptococcus pneumoniae, a pathogen of humans, was isolated upon CSF microbiological culture. Treatment consisted of intravenous antibiotics, supportive care, and anticonvulsants for the generalized seizures which developed shortly after admission. The dog responded to therapy and two years later exhibited only a mild, residual head tilt. The pathogenesis and treatment of bacterial meningoencephalitis in dogs are reviewed.

Molecular characterization of equine isolates of Streptococcus pneumoniae: natural disruption of genes encoding the virulence factors pneumolysin and autolysin.

Although often considered a strict human pathogen, Streptococcus pneumoniae has been reported to infect and cause pneumonia in horses, although the pathology appears restricted compared to that of human infections. Here we report on the molecular characterization of a group of S. pneumoniae isolates obtained from horses in England and Ireland. Despite being obtained from geographically distinct locations, the isolates were found to represent a tight clonal group, virtually identical to each other but genetically distinguishable from more than 120 divergent isolates of human S. pneumoniae. A comprehensive analysis of known pneumococcal virulence determinants was undertaken in an attempt to understand the pathogenicity of equine pneumococci. Surprisingly, equine isolates appear to lack activities associated with both the hemolytic cytotoxin pneumolysin, often considered a major virulence factor of pneumococci, and the major autolysin gene lytA, also considered an important virulence factor. In support of phenotypic data, molecular studies demonstrated a deletion of parts of the coding sequences of both lytA and ply genes in equine pneumococci. The implications of these findings for the evolution and pathogenicity of equine S. pneumoniae are discussed.

Serodiagnosis of Streptococcus pneumoniae infection in guinea pigs by an enzyme-linked immunosorbent assay.

Guineapig antibodies to Streptococcus pneumoniae (SPN) serotype 19F were detected by an enzyme-linked immunosorbent assay using a simple procedure. In experimentally infected hosts, antibody was detectable as early as 2 to 3 weeks after infection, and high titres were maintained for a long period. Antibodies higher than 1:64 were regarded as specific. In a field study, high antibody titres were shown in SPN enzootic colonies in contrast to negative or low antibody titres in a majority of the animals from non-enzootic and SPF colonies.

Inapparent Streptococcus pneumoniae type 35 infections in commercial rats and mice.

Streptococcus pneumoniae was isolated from specific-pathogen-free rodents in two rooms at a commercial breeding facility during vendor surveillance testing. In a survey of 274 animals from the two rooms over a period of 7 months, capsular serotype 35 S. pneumoniae was isolated from the upper respiratory tracts of 11% (9 of 82) of C57BL/6 mice in room A and 14% (10 of 72) of F344 rats in room B, but not from WKY rats, BALB/c mice or DBA/2 mice from room A. In both C57BL/6 mice and F344 rats, older rodents had higher colonization frequencies. Nasal lavage cultures gave the best results in identifying colonized rodents. No clinical illness or microscopic lesions were associated with pneumococcal colonization in rats or mice, and no other evidence of potential pathogen infection was found except for positive serologic tests for mouse rotavirus in mice. This is the first report of natural pneumococcal infection in mice, and the first report of type 35 S. pneumoniae infection in rodents. The findings support an earlier observation that pneumococcal infections in rat colonies tend to be monotypic and suggest that the same may be true in mice.

Septicemia and septic arthritis caused by Streptococcus pneumoniae in a cat: possible transmission from a child.

Streptococcus pneumoniae serotype 23F was isolated from the blood and synovial fluid of an acutely ill, 15-year-old castrated male cat and from the nasopharynx of that regularly played with it, an infant child. Information presented supports the hypothesis that the infection was transmitted from child to cat.

Isolation of Streptococcus pneumoniae from the respiratory tract of horses.

Streptococcus pneumoniae was isolated from nasopharyngeal swabs and tracheal washings taken from Thoroughbred horses in training at three of four separate stables that were sampled during investigations into respiratory disease. The growth of Strep pneumoniae in culture was enhanced by an environment enriched with carbon dioxide. In one stable, five of 15 horses that were sampled repeatedly were found to carry the organism for at least four months. There was an apparent association between lower respiratory tract inflammatory disease and heavy growths (10(6) to 10(8) colony forming units/ml) predominantly of Strep pneumoniae or of that organism together with large numbers of Strep zooepidemicus obtained from tracheal washings. Twelve strains of Strep pneumoniae isolated from three stables were all of capsule Type 3. Only one strain, which was of capsule Type 9, was isolated from nose and throat swabs taken from 32 staff working in one of the stables and suggested an absence of cross infection between horses and their handlers in this instance.