Opportunistic Infections (Infectious Disease (Totowa, N.J.).)
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Garden bird feeding practice in Great Britain has altered over recent years with increased adoption of summer feeding and increased provision of sunflower and niger seed which might have led to increased concentration of birds at feeders. Trichomonas gallinae can be transmitted through direct contact between birds, for example courtship and feeding of young, and through indirect routes including shared food and water sources  ,  , however further studies are required to assess the relative importance of these transmission routes for finches.
Establishing the nature and frequency of disease transmission at garden feeders is thus clearly important to identify if mitigation measures are required and, if so, how they should be employed. The greenfinch and chaffinch are both common garden bird visitors in England and Wales across the year, ranking number 9 and 10 in the most frequent garden visitors in the GBW scheme . Both species are gregarious, visiting gardens in flocks, but other granivorous passerine species are reported in a comparable number of gardens around feeding stations, for example house sparrows, great tits and blue tits, and they also feed in groups.
More generally, T. Continued monitoring of diseases in wild bird populations is required to better quantify and understand their impact on population dynamics  ,  and to identify future changes in host-parasite relationships. No live animals were used for this research, however, the project was reviewed and approved by the Zoological Society of London's Ethics Committee.
The GBHi was established before the emergence of trichomonosis in British finches was identified and has always included wide-ranging investigations to identify pathogens responsible for garden bird disease. The GBHi surveillance of garden bird morbidity and mortality takes place via opportunistic reports obtained from the general public and weekly reports from identified volunteers who form a systematic reporting network.
As opportunistic reports are vulnerable to temporal or spatial observer bias, for example following regional media reports, the systematic surveillance network was established in April in order to quantify disease prevalence rates.
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A random sample of 1, volunteers stratified by the number of participants in each recording region were approached with a view to recording additional information relating to the incidence of diseased and dying birds in their gardens. Post mortem examinations using a standardised protocol were performed on carcasses submitted from a subset of reported garden bird mortality incidents. Cases were selected for post-mortem investigation based on fresh carcass availability.
Birds thought to have died as a result of trauma, predation or infectious disease were examined; our selection criteria did not specifically or solely target finch species or suspected cases of trichomonosis, but rather aimed to achieve a representative cross-section of species and aetiologies. Each submitted carcass was assigned a unique post mortem reference code and the species, age, sex and body weight were recorded for each bird examined. Systematic external and internal examinations of body systems were performed and any gross lesions described.
Where indicated, and where the state of carcass decomposition permitted, samples were taken for microbiological, parasitological and histopathological investigations. Liver and contents of the mid-small intestinal loop were sampled aseptically from the majority of cases, as were any lesions found, and were examined for the presence of pathogenic bacteria. The same bacteriology protocol was used for examination of necrotic ingluvitis lesions as for the intestinal contents with the exception of the modified CCDA-Preston media. Bacterial isolates were identified using colony and Gram's staining morphology, followed by biochemical properties which were determined using the API biochemical test strip method API-BioMerieux, Marcy l'Etoile, France.
Oxoid, UK and screened for motile trichomonads at 24, 48, 72 hrs and 5 days. Wet mount preparations of small intestinal contents were examined in a subset of cases for evidence of nematode, cestode and protozoan parasites. A combination of morphological and molecular techniques was used to identify the trichomonad species. Giemsa-stained preparations of trichomonad cultures were examined using light microscopy to assess parasite morphology. These were prepared by placing a drop of active trichomonas culture onto a standard glass microscope slide; this was then air dried, alcohol-fixed and stained using routine methods.
Transmission and scanning electron microscopy was performed on trichomonad cultures fixed in 2.
DNA was extracted from trichomonad cultures using the same technique. Oligonucleotide primers were supplied by Operon Biotechnologies, Germany. Each PCR run contained a negative control of water and a positive control of purified trichomonad DNA obtained from parasites cultured from an affected greenfinch found dead as part of this study.
Chromatograph profiles were inspected using Chromas 2 software www. A nested PCR protocol was designed to increase the sensitivity of detection of trichomonad parasite DNA in template DNA extracted from lesions sampled post mortem and to provide a diagnostic tool for cases where autolysed carcass condition precluded Trichomonas sp. The PCR product nucleotide sequence was obtained from a pure trichomonad culture obtained from an affected greenfinch.
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The reaction mixture for the second amplification was the same as for the first, except for the use of the nested primers. Each amplification contained a negative control, consisting of water and a positive control of purified DNA obtained from cultured trichomonad parasites from a greenfinch. Chromatograph profiles were inspected using Chromas 2 software. Reliable discrimination between necrotic ingluvitis due to salmonellosis or trichomonosis in finches is not possible based on gross examination alone.
In order to evaluate whether our nested PCR cross-reacted non-specifically with Salmonella Typhimurium Definitive Type DT 40 and DT56 variant v , lesions from 56 greenfinches and six chaffinches confirmed to have died due to salmonellosis by microbiological examination in and Lawson, unpublished data were screened. This indicates that 8 cases examined outside the study period might have had concurrent infection with Trichomonas sp.
Carcass condition precluded parasite culture in all but one of these cases, but this case yielded trichomonad parasites confirming the existence of dual infection. These findings suggest that the nested PCR does not cross react non-specifically with Salmonella Typhimurium DT40 and DT56 v and can be used within our case definition for the diagnosis of trichomonosis in finches. Salmonellosis also causes necrotic ingluvitis and this was diagnosed by lesions being positive for Salmonella sp. No cases of dual infection were identified during the study period.
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Garden Bird Watch participants c. Participants maintain a consistent level of observational effort from one week to the next; data from weeks that are under- or over-observed are discarded. Variation in observer effort and competence is inevitable, but this can be controlled for by introducing a site effect into the models used to examine the data . Almost all the participants provide food of some kind the range of food provided for garden birds is also recorded on a weekly basis for wild birds and feeding stations are generally the focal point of the study areas.
The subset of participants that took part in GBH i surveyed all or part of their garden systematically in a consistent manner each week to record the number, and putative cause, of dead or sick birds found. The clinical signs of ill health in birds affected by trichomonosis typically included non-specific malaise, although dysphagia was noted in a large proportion of reported incidents. This contrasts with the clearly recognisable external signs of conjunctivitis caused by Mycoplasma gallisepticum in the house finch and which were used to monitor spatial spread of mycoplasmosis .
In order to evaluate the geographical distribution of the trichomonosis epidemic, data from the opportunistic and systematic reporting schemes were examined for the period between 1 April and 30 September This interval was selected to minimise the likelihood of confounding these data with mortality due to salmonellosis, outbreaks of which occur during the winter months and which also result in non-specific signs of malaise in finches.
Previous studies of salmonellosis in Great Britain  , continental Europe  and North America  along with our own observations on salmonellosis in passerines over the period — Lawson and Cunningham, unpublished data , have shown the disease to be seasonal, occurring almost exclusively within the period 1 st October to 31 st March.
As resources and logistics did not allow all dead birds found to be submitted for post mortem examination, we established specific criteria for determining if a mortality incident should be classified as likely being due to trichomonosis. For the purposes of this analysis, a trichomonosis incident was defined if, within the six month period of the study between 1 April and 30 September , mortality included two or more dead finches greenfinch or chaffinch , one or more sick finch es with typical signs of disease, or if trichomonosis was confirmed post mortem.
To give a measure of incidence for the opportunistic reports, we expressed the total number of trichomonosis incidents reported in each county per thousand households according to the UK National Census . Each week, GBW participants recorded the presence of each bird species in their garden and, optionally, the peak number of birds counted each week . For this analysis we used the gardens for which data were submitted in every week from January to June in England south of We modelled reporting rates using generalised additive or linear mixed models, fitted using the gamm function in package mgcv for R 2.
To exclude the possibility that changes in other environmental factors, such as climate, between years might have caused changes in greenfinch numbers, we also modelled reporting rates of chaffinch, which has a similar body size and ecology to the greenfinch, and was the second most frequently affected species in which trichomonosis was recorded, and of dunnock, which also feeds around garden feeders  but in which trichomonosis was rarely recorded only three cases diagnosed in and combined.
Both species show similar patterns of spatial and temporal abundance in gardens to the greenfinch . We predicted that, if trichomonosis was responsible for changes in numbers, chaffinch should show an identifiable, but less marked, response and dunnock should show little response; if other factors, such as climatic factors or a change in resource availability, were involved then the response among the three species should be similar.
We used the default level of smoothing, as increasing the potential for smoothing by changing the basis, k did not alter the results materially. To account for variation among gardens in the probability of birds being present and observer ability we included garden identifier as a random-effect term. Preliminary models including garden identifier as a fixed effect showed the distribution of these effects, as for other large-scale schemes run by BTO, were indeed approximately normally distributed.
The weekly records of bird presence are likely to exhibit serial auto-correlation, both temporally and spatially. This may reflect the fact that distance between gardens was generally much greater than the ambit of individual foraging flocks and it is unlikely the provisioning behaviour in any one sample garden influenced the behaviour in neighbouring sample gardens. We therefore included an AR1 auto-regressive correlation structure with weeks numbered consecutively through the entire period to account for the correlation between records in consecutive weeks; the degree of auto-correlation was assumed to be the same for each garden.
To quantify impacts on the breeding season following the epidemic of trichomonosis in autumn , we constructed a model to compare reporting rate at the start of the breeding season weeks 13—21, 26 March—28 May with the average reporting rate for the same period in the previous two years which, as far as could be determined, were typical.
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We restricted the analysis to the previous two years to avoid potential confounding effects of long-term trends in bird numbers and reporting rates. The estimate of this latter term then gives the change in reporting rate in relative to that in the previous two years. As before, we included garden identifier as a random effect and fitted an auto-correlated binomial error structure.
Mortality due to trichomonosis varied spatially throughout the country, so we defined three regions representing areas of High, Intermediate and Low incidence of trichomonosis, based on the results of opportunistic sampling. We restricted these analyses to England south of a line from the Mersey to the Humber approx. To test for differences in the change in reporting rate in spring among areas, we included an interaction term between area and the dummy year variable described above.
Numbers of birds present across Britain during the breeding season are monitored using line-transect counts in a sample of c. BBS transects are undertaken in all habitats, rather than being restricted to gardens as with the GBW scheme. An index of relative abundance based on a generalized linear Poisson model with categorical site and year fixed-effects is produced annually; we obtained indices for the three county groupings to measure the relative change in breeding population between and in each region.
Ball and the referees for their helpful comments on the manuscript. Analyzed the data: RR. Modelling of data: RR.
new.urbanreef.com/fuma-top-cell-phone.php Development of new molecular analytic tools: OCH. Technical assistance: MWP. Supervision of molecular analyses: KT. Browse Subject Areas?
Click through the PLOS taxonomy to find articles in your field. Abstract Emerging infectious diseases are increasingly cited as threats to wildlife, livestock and humans alike. Introduction Emerging infectious diseases EIDs are increasingly cited as threats to wildlife, livestock and humans alike  and can be a major threat to geographically isolated or critically endangered wild bird populations  , .
Download: PPT. Figure 1. Seasonal incidence of opportunistic reports in all garden bird mortality — Identification of the disease organism Necrotic ingluvitis, typically extending through the full thickness of the oesophageal wall and often involving adjacent connective tissue, was diagnosed through post-mortem examination Fig. Figure 2. Individuals at higher risk are often prescribed prophylactic medication to prevent an infection from occurring.
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A patient's risk level for developing an opportunistic infection is approximated using the patient's CD4 T-cell count and sometimes other markers of susceptibility. Common prophylaxis treatments include the following: .