Bergeyella zoohelcum isolated from oral cavities of therapy dogs.

Bergeyella zoohelcum causes rare but severe human clinical diseases, which mostly arise from animal bites. Notably, Bergeyella infections can also occur in older people after prolonged exposure to dogs or cats without biting. We detected B. zoohelcum in oral cavities of therapy dogs in close contact with older people residing in nursing homes. Twenty-two bacterial isolates were identified as B. zoohelcum by using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) and 16S rRNA gene sequencing. Our results showed that MALDI-TOF MS is an effective tool for rapid identification of rarely isolated, difficult-to-identify microorganisms, such as B. zoohelcum, derived from not only human clinical samples but also animal samples. To our knowledge, this is the first report on detection of B. zoohelcum from therapy dogs. We have provided information on dog-assisted therapy to improve the relationship between humans and animals in ageing societies, particularly for preventive healthcare of older people living in nursing care facilities.

A cluster of Chryseobacterium indologenes cases related to drainage water in intensive care units.

In this outbreak, 12 patients in intensive care units acquired a Chryseobacterium indologenes infection. Cultures from sinkholes and air samples were positive for C. indologenes. After removing wash basins, no new cases appeared. Sinkholes, potentially contaminated, can act as a reservoir for C. indologenes and other microorganisms. Thus, patients and equipment should be protected from sink splashes to avoid contamination.

Pathogenic Elizabethkingia miricola Infection in Cultured Black-Spotted Frogs, China, 2016.

Multiregional outbreaks of meningitis-like disease caused by Elizabethkingia miricola were confirmed in black-spotted frog farms in China in 2016. Whole-genome sequencing revealed that this amphibian E. miricola strain is closely related to human clinical isolates. Our findings indicate that E. miricola can be epizootic and may pose a threat to humans.

Mini review: New pathogen profiles: Elizabethkingia anophelis.

Within a little more than 5 years since its taxonomic description in 2011, from the midgut of mosquitoes, Elizabethkingia anophelis has emerged as an important causes of sepsis in adults and children and in cases of neonatal meningitis. At least 3 moderate- to large-scale outbreaks of disease have been caused by this bacterium, the largest 2 occurring in the Midwest United States in 2015-2016. Several studies suggest that E. anophelis, and not E. meningoseptica, is the predominant human pathogen of this genus; identification to species is difficult. Little is presently known regarding its epidemiology, modes of transmission, and pathogenicity as it relates to virulence-associated factors.

Evidence for Elizabethkingia anophelis transmission from mother to infant, Hong Kong.

Elizabethkingia anophelis, recently discovered from mosquito gut, is an emerging bacterium associated with neonatal meningitis and nosocomial outbreaks. However, its transmission route remains unknown. We use rapid genome sequencing to investigate 3 cases of E. anophelis sepsis involving 2 neonates who had meningitis and 1 neonate's mother who had chorioamnionitis. Comparative genomics revealed evidence for perinatal vertical transmission from a mother to her neonate; the 2 isolates from these patients, HKU37 and HKU38, shared essentially identical genome sequences. In contrast, the strain from another neonate (HKU36) was genetically divergent, showing only 78.6% genome sequence identity to HKU37 and HKU38, thus excluding a clonal outbreak. Comparison to genomes from mosquito strains revealed potential metabolic adaptations in E. anophelis under different environments. Maternal infection, not mosquitoes, is most likely the source of neonatal E. anophelis infections. Our findings highlight the power of genome sequencing in gaining rapid insights on transmission and pathogenesis of emerging pathogens.

Investigation of the link between broodstock infection, vertical transmission, and prevalence of Flavobacterium psychrophilum in eggs and progeny of Rainbow Trout and Coho Salmon.

The etiological agent of bacterial coldwater disease (BCWD), Flavobacterium psychrophilum, can be transmitted both vertically and horizontally. Outbreaks of BCWD can result in significant losses in salmonid aquaculture. Reduction of outbreaks in fry may be possible through implementation of a management strategy in which progeny of heavily infected broodstock are culled from the general population. Diagnostic assays to quantify F. psychrophilum concentrations in tissue samples and confirm presence of the bacterium in ovarian fluid have been previously validated. In the current study, these assays were used to screen 60 female Rainbow Trout Oncorhynchus mykiss and 60 female Coho Salmon O. kisutch broodstock at two aquaculture facilities. Eyed eggs from 10 female broodstock (five fish from each facility) exhibiting graded levels of infection were transferred to the University of Idaho and monitored through early life stages for the presence of F. psychrophilum. Female Rainbow Trout broodstock were not positive for F. psychrophilum by enzyme-linked immunosorbent assay (ELISA) and prevalence was low in these progeny. However, ELISA optical density values for kidney correlated to F. psychrophilum prevalence in progeny (r = 0.938, P < 0.05) of Coho Salmon. Nested PCR on ovarian fluid was not a reliable indicator of vertical transmission in either species as broodstock ovarian fluid results did not correlate to F. psychrophilum prevalence in eyed eggs. Further research with these assays is necessary; however, results from this study indicate that broodstock screening may be a potential tool for evaluating F. psychrophilum infection levels, which could become an important component for disease management.

Improved broiler performance associated with Ornithobacterium rhinotracheale vaccination in breeders.

During a field study in 2010 the daily growth, feed conversion, first-week mortality, broiler loss due to mortality and slaughterhouse condemnation, and production index were monitored in 100 broiler flocks derived from four breeder farms vaccinated with Nobilis OR inac and four Ornithobacterium rhinotracheale-unvaccinated breeder farms of the same organization in Belgium. Other parameters related to the broiler flocks, such as flock size, season, age of the breeders, and corresponding breeder farms, were also noted. All gathered data were examined with ANOVA, linear correlation, and linear regression analyses. Results demonstrated a significant 22.3% lower broiler loss and a significant 3.9% higher production index in the broiler flocks derived from breeders vaccinated with Nobilis OR inac. These results confirm field observations obtained in 1999, thereby providing further evidence for an effect of O. rhinotracheale vaccination in breeders with regard to the improved performance of broilers.

Intensive fish farming and the evolution of pathogen virulence: the case of columnaris disease in Finland.

Ecological changes affect pathogen epidemiology and evolution and may trigger the emergence of novel diseases. Aquaculture radically alters the ecology of fish and their pathogens. Here we show an increase in the occurrence of the bacterial fish disease Flavobacterium columnare in salmon fingerlings at a fish farm in northern Finland over 23 years. We hypothesize that this emergence was owing to evolutionary changes in bacterial virulence. We base this argument on several observations. First, the emergence was associated with increased severity of symptoms. Second, F. columnare strains vary in virulence, with more lethal strains inducing more severe symptoms prior to death. Third, more virulent strains have greater infectivity, higher tissue-degrading capacity and higher growth rates. Fourth, pathogen strains co-occur, so that strains compete. Fifth, F. columnare can transmit efficiently from dead fish, and maintain infectivity in sterilized water for months, strongly reducing the fitness cost of host death likely experienced by the pathogen in nature. Moreover, this saprophytic infectiousness means that chemotherapy strongly select for strains that rapidly kill their hosts: dead fish remain infectious; treated fish do not. Finally, high stocking densities of homogeneous subsets of fish greatly enhance transmission opportunities. We suggest that fish farms provide an environment that promotes the circulation of more virulent strains of F. columnare. This effect is intensified by the recent increases in summer water temperature. More generally, we predict that intensive fish farming will lead to the evolution of more virulent pathogens.

Saprophytism of a fish pathogen as a transmission strategy.

Fish farming creates conditions where disease transmission is enhanced and antibiotic treatments are commonly used to cure bacterial diseases to prevent severe losses due to infections. Ability to persist in such an environment has been suggested to lead to the evolution of high virulence. Columnaris disease caused by Flavobacterium columnare is a growing problem in freshwater fish farming. Transmission of the disease is poorly known, and survival of F. columnare in the rearing environment has not been studied. This paper addresses both transmission of columnaris disease and survival strategy of F. columnare. Saprophytic activity of F. columnare was studied by infecting rainbow trout fingerlings before and immediately after death and by following bacterial shedding from the fish carcasses. From fish killed immediately after infection, bacteria were shed at high rates for 5 days, and from fish exposed to F. columnare post mortem for 8 days. In another experiment, rainbow trout fingerlings were experimentally infected with F. columnare and monitored for transmission of the bacteria post infection until and after the death of the fish. The transmission of columnaris disease to living rainbow trout was the most efficient from dead fish, from which bacteria were shed into water at higher rates than from living fish. We also found that F. columnare can survive at least for 5 months in both sterilized distilled and lake water. These results show that death of the host causes no cost for F. columnare; it thrives in alive and dead fish, and in water. Saprophytism may have been a transition stage to pathogenicity of this originally harmless water bacterium, and maintained as an effective transmission and survival strategy of F. columnare. Our findings also suggest that F. columnare may be able to persist in the rearing environment during antibiotic treatments of the living fish.

Susceptibility of zebra fish Danio rerio to infection by Flavobacterium columnare and F. johnsoniae.

Flavobacterium columnare is a serious pathogen in a wide range of fish species. F. johnsoniae is an opportunistic pathogen of certain fish. Both are gliding bacteria. These species were tested for their ability to infect the zebra fish Danio rerio. Both injection and bath infection methods were tested. The results indicate that F. johnsoniae is not an effective pathogen in D. rerio, but that F. columnare is an effective pathogen. F. johnsoniae did not cause increased death rates following bath infection, but did cause increased death rates following injection, with an LD50 (mean lethal dose) of approximately 3 x 10(10) cfu (colony-forming units). Non-motile mutants of F. johnsoniae produced a similar LD50. F. columnare caused increased death rates following both injection and bath infections. There was considerable strain variation in LD50, with the most lethal strain tested producing an LD50 of 3.2 x 10(6) cfu injected and 1.1 x 10(6) cfu ml(-1) in bath experiments, including skin damage. The LD50 of F. columnare in zebra fish without skin damage was > 1 x 10(8), indicating an important effect of skin damage.

Influence of rearing conditions on Flavobacterium columnare infection of rainbow trout, Oncorhynchus mykiss (Walbaum).

The influence of rearing conditions on Flavobacterium columnare infection of rainbow trout, Oncorhynchus mykiss (Walbaum), was studied experimentally in the laboratory and at a fish farm. In experiment I, the effect of parasitic infection on columnaris disease was studied using F. columnare carrier fish. The fish were exposed to Diplostomum spathaceum cercariae and a set of other stressors in order to induce clinical columnaris infection. Parasitic infection and other stressors failed to induce the disease. Disease occurred when the fish were challenged with F. columnare, but D. spathaceum infection did not enhance the severity of the infection. In experiment II, the influence of rearing density and water temperature was studied. Overall mortality was highest in fish at normal rearing density with high temperature (+23 degrees C). At low temperature (+18 degrees C) mortality was not affected by rearing density, but the transmission of columnaris disease was faster at normal rearing density at both temperatures. This supports the view that reduction of fish density could be used in prevention of columnaris disease especially if water temperature is high. Because the lower rearing density can also decrease the transmission of ectoparasites and penetrating endoparasites, it could be an efficient tool in ecological disease management.

Transmission and detection of Flavobacterium columnare in channel catfish Ictalurus punctatus.

A specific and rapid PCR detection method for Flavobacterium columnare based on the 16S-23S rDNA intergenic spacer region (ISR) of the ribosomal RNA operon has been developed. The ISR of 30 F. columnare strains and other Flavobacterium species was amplified using universal primers and sequenced. Once F. columnare specific sequences within the ISR were recognized, specific PCR primers were designed against them (FCISRFL and FCISRR1). The primers were sensitive and able to detect as low as 7 colony forming units from pure culture by PCR. The new PCR detection method was applied to experimentally infected channel catfish. Two different experiments in which channel catfish fingerlings were infected by intramuscular injection or by immersion bath showed the advantage of the PCR method over standard culture techniques. F. columnare was detected by PCR in both tank water and catfish tissue samples with a higher frequency and in less time than standard microbiological methods. Furthermore, PCR detection confirmed that F. columnare can be transmitted horizontally indirectly through the water column without fish-to-fish contact. The newly developed PCR detection method for F. columnare was more sensitive and rapid than standard culture on bacteriological media for detection of F. columnare in channel catfish tissues and in tank water.

The effect of route of inoculation and challenge dosage on Riemerella anatipestifer infection in Pekin ducks (Anas platyrhynchos).

Riemerella anatipestifer is a gram-negative bacteria that can cause disease in a wide variety of wild and domesticated birds, especially waterfowl. The infection can be peracute, acute, or chronic. Although various routes of transmission have been proposed, to date, there is little information on the effects of route of transmission and challenge dosage on R. anatipestifer infection. Hence, the objective of this study was to determine the effect of route of inoculation and challenge dosage on R. anatipestifer infection and pathology. To achieve this objective, one hundred forty-seven 14-day-old white Pekin ducks (Anas platyrhynchos) were equally divided into 13 experimental groups (12 challenge and 1 control group). Each challenge group had 11 ducks. The control group had 15 ducks. Four routes of inoculation were evaluated (intranasal, oral, subcutaneous, and intravenous). Three dosage levels were evaluated for each inoculation route (10(2), 10(4), and 106 colony forming units [CFU]/ml). At the 106 CFU/ml dosage level, mortality was most associated with the subcutaneous (91%) and intravenous (82%) routes, followed by the nasal (18%) and oral (9%) routes. A unique pathologic lesion was found in the bursa of Fabricius and spleen of affected birds. Within the spleen and bursa of Fabricius, there were varying degrees of lymphoid depletion and necrosis within the cortical and medullary regions. These pathologic lesions have not been previously reported in ducks with R. anatipestifer infection.

Studies of the transmission routes of Ornithobacterium rhinotracheale and immunoprophylaxis to prevent infection in young meat turkeys.

The importance and prevention of the horizontal as well as the vertical transmission of Ornithobacterium rhinotracheale were investigated. In our first experiment we observed that specific-pathogen-free broiler chickens that were placed in hatching incubators at a commercial turkey hatchery during hatch showed respiratory tract lesions at postmortem examination that were positive for O. rhinotracheale by bacteriology and immunohistology. It appeared that vertical transmission occurred and that horizontal transmission of O. rhinotracheale is possible. In a second experiment, the turkeys derived from vaccinated parents showed significantly fewer respiratory tract lesions at postmortem examination at 16 days of age than the birds derived from nonvaccinated parents. In a third experiment, all vaccinated young birds, regardless of the vaccination state of their parents, showed significantly fewer respiratory tract lesions at 6 wk of age. We concluded that vaccination of the breeders reduces vertical transmission and that vaccination of the progeny is needed to resist challenge at 6 wk of age.

Flavobacterium psychrophilum infections in salmonid fish.

Flavobacterium psychrophilum is the causative agent of bacterial cold water disease and rainbow trout fry syndrome, disease entities responsible for substantial economic losses in salmonid aquaculture. Problems associated with epizootics include high mortality rate, increased susceptibility to other diseases, high labour costs of treatment and the enormous expenditure on chemotherapy. Despite the increasing significance of the disease, the pathogenesis of F. psychrophilum infections has only been partially elucidated, hampering the development of preventive measures to efficiently combat this disease condition. This literature review discusses the agent and the disease it causes, with emphasis on the bacterium-host interactions.