Seasonal Variation in Detection of Haemosporidia in a Bird Community: A Comparison of Nested PCR and Microscopy.

In a 2-yr study on prevalence of Haemosporidia in an avian community in Ithaca, New York, USA, we tested the hypothesis that apparent seasonal variation in prevalence is influenced by the detection protocol. We confirmed a higher detection of Haemosporidia using a molecular diagnosis technique (PCR) than by microscopy; this further increased when the PCR test was triplicated. Microscopic examination and PCR techniques have different specificity and sensitivity and therefore different probabilities of detecting hemoparasites. Birds with chronic infections or sampled during winter often have very low parasitemia, and such infections may be missed by microscopy but detected by PCR. Haemosporidian prevalence was higher during the breeding season than during the nonbreeding season regardless of the method used. Detection of Leucocytozoon spp. infection from blood smears using microscopy was challenging.

Are Purple Finches (Haemorhous purpureus) the Next Host for a Mycoplasmal Conjunctivitis Epidemic?

Ever since 1994, when the bacterial pathogen Mycoplasma gallisepticum jumped from poultry to wild birds, it has been assumed that the primary host species of this pathogen in wild North American birds was the house finch (Haemorhous mexicanus), in which disease prevalence was higher than in any other bird species. Here we tested two hypotheses to explain a recent increase in disease prevalence in purple finches (Haemorhous purpureus) around Ithaca, New York. Hypothesis 1 is that, as M. gallisepticum evolved and became more virulent, it has also become better adapted to other finches. If this is correct, early isolates of M. gallisepticum should cause less-severe eye lesions in purple finches than in house finches, while more-recent isolates should cause eye lesions of similar severity in the two species. Hypothesis 2 is that, as house finch abundance declined following the M. gallisepticum epidemic, purple finches around Ithaca increased in abundance relative to house finches and purple finches are thus more frequently exposed to M. gallisepticum-infected house finches. This would then lead to an increase in M. gallisepticum prevalence in purple finches. Following an experimental infection with an early and a more-recent M. gallisepticum isolate, eye lesions in purple finches were more severe than in house finches. This did not a support Hypothesis 1; similarly, an analysis of Project Feeder Watch data collected around Ithaca did not show differences in changes in purple and house finches' abundance since 2006, a result which does not support Hypothesis 2. We conclude that purple finch populations will, unlike those of house finches, not suffer a severe decline because of a M. gallisepticum epidemic.

¿Son los pinzones purpúreos (Haemorhous purpureus) los próximos huéspedes de una epidemia de conjuntivitis por micoplasma? Desde el año 1994, cuando el patógeno bacteriano Mycoplasma gallisepticum saltó de las aves comerciales a las aves silvestres, se ha supuesto que la principal especie huésped de este patógeno en las aves silvestres de América del Norte era el pinzón mexicano (Haemorhous mexicanus), en el que la prevalencia de la enfermedad era mayor que en cualquier otra especie aviar. En este estudio se analizaron dos hipótesis para explicar un aumento reciente en la prevalencia de la enfermedad en los pinzones purpúreos (Haemorhous purpureus) alrededor de Ithaca, en Nueva York. La hipótesis 1 es que, a medida que M. gallisepticum evolucionó y se volvió más virulento, también se adaptó mejor a otros pinzones. Si esto es correcto, los aislamientos tempranos de M. gallisepticum deberían causar lesiones oculares menos graves en los pinzones purpúreos que en los pinzones mexicanos, mientras que los aislamientos más recientes deberían causar lesiones oculares de gravedad similar en las dos especies. La hipótesis 2 es que, a medida que la abundancia de pinzones mexicanos disminuyó después de la epidemia de M. gallisepticum, los pinzones purpúreos alrededor de Ithaca aumentaron en abundancia en relación con los pinzones mexicanos y, por lo tanto, los pinzones morados están expuestos con mayor frecuencia a los pinzones caseros infectados con M. gallisepticum. Esto conduciría a un aumento de la prevalencia de M. gallisepticum en los pinzones purpúreos. Después de una infección experimental con un aislamiento temprano y uno más reciente de M. gallisepticum, las lesiones oculares en los pinzones purpúreos fueron más graves que en los pinzones mexicanos. Esto no apoyó la Hipótesis 1; de manera similar, un análisis de los datos del Proyecto Feeder Watch recopilados alrededor de Ithaca no mostró diferencias en los cambios de la abundancia de pinzones purpúreos y mexicanos desde 2006, un resultado que no respalda la Hipótesis 2. Se concluye que las poblaciones de pinzones purpúreos, a diferencia de las de los pinzones mexicanos, no sufrieron un declive severo a causa de una epidemia de M. gallisepticum.

CHANGES IN TISSUE TROPISM OF MYCOPLASMA GALLISEPTICUM FOLLOWING HOST JUMP.

Mycoplasma gallisepticum, a pathogen of worldwide economic importance in poultry, is recovered in chickens, especially from the respiratory tract. Some strains, however, are specialized to other tissues and because it jumps from poultry to wild birds, the new strains also cause severe conjunctivitis in new hosts. Nevertheless, most studies of M. gallisepticum in wild birds use choanal swabs or combine choanal and conjunctival swabs to quantify bacterial load. Because the clinical signs associated with M. gallisepticum infection differ markedly between poultry and House Finches (Haemorhous mexicanus), we compared the bacterial load in choanal and conjunctival samples following experimental inoculation of House Finches with M. gallisepticum isolates originating from poultry or from House Finches. This allowed us to test two hypotheses: M. gallisepticum changed tissue tropism, or M. gallisepticum simply expanded its within-host niche. By comparing bacterial loads from choanal and conjunctival swabs in birds inoculated with one of a suite of M. gallisepticum isolates, we found support for hypothesis 2. The choanal loads in House Finches did not differ between isolates, while the conjunctival loads of birds inoculated with poultry isolates were lower than in birds inoculated with House Finch isolates. When measuring the bacterial load of M. gallisepticum in birds, it is important to sample and analyze separately choanal and conjunctival swabs, as quantifying bacterial loads in pooled samples may not provide reliable information on differences in virulence.

THE EFFECT OF DEXAMETHASONE ON HEMATOLOGIC PROFILES, HEMOSPORIDIAN INFECTION, AND SPLENIC HISTOLOGY IN HOUSE FINCHES (HAEMORHOUS MEXICANUS).

Research on host response to infectious disease often involves pharmacological induction of immunosuppression, frequently through administration of dexamethasone. Reports on the effect of dexamethasone in birds are largely restricted to poultry and pigeons. This study describes changes in white blood cell (WBC) differentials, hemoparasite counts, splenic histology, and splenic CD3 immunoreactivity in House Finches (Haemorhous mexicanus). Experimental group birds (n=9) were treated with a daily intramuscular injection of 25 µg of dexamethasone for 8 d; a control group (n=9) received daily saline solution. Smears were made with blood collected immediately before the first dose (day 0) and on d 4, 8, and 9, and stained with modified Wright. The WBC differential counts were performed by three blinded observers, parasite counts by two blinded observers, and histology by one blinded observer. Dexamethasone-treated birds experienced relative heterophilia and lymphopenia on d 4 (P=0.008); heterophilia was also present at d 8 (P=0.018). Hemosporidian counts were significantly increased in dexamethasone-treated birds on d 4 and 8 (P=0.048 and P=0.031, respectively). In contrast with control birds, all dexamethasone-treated birds lacked histologically apparent splenic lymphoid follicles (P<0.001). No significant difference was observed in splenic CD3 immunoreactivity between groups. Our results indicate that dexamethasone has an effect on the hematologic profile of House Finches and suggest that it may be a useful method to induce immunosuppression in this species.

Ectoparasitism during an avian disease outbreak: An experiment with Mycoplasma-infected house finches and ticks.

Hosts are typically co-parasitized by multiple species. Parasites can benefit or suffer from the presence of other parasites, which can reduce or increase the overall virulence due to competition or facilitation. Outcomes of new multi-parasite systems are seldom predictable. In 1994 the bacterium Mycoplasma gallisepticum jumped from poultry to songbirds in which it caused an epidemic throughout North America. Songbirds are often parasitized by hard ticks, and can act as reservoirs for tick-borne pathogens. We tested the hypothesis that Mycoplasma infection in house finches influences North America's most important tick vector Ixodes scapularis, by affecting the tick's feeding success, detachment behaviour and survival to the next stage. Most ticks detached during the daylight hours irrespective of the bird's disease status and time since infestation. Birds incrementally invested in anti-tick resistance mechanisms over the course of the experiment; this investment was made earlier in the Mycoplasma-infected birds. At higher tick densities, the feeding success on birds with more severe conjunctivitis was lower than in the uninfected birds. Throughout the experiment we found positive density dependent effects on the tick's feeding success. More diseased hosts suffered more from the tick infestations, as shown by reduced haematocrits. Three Mycoplasma-infected birds died during the weeks following the experiment, although all birds were kept in optimal housing conditions. Mycoplasma made the bird a less accessible and valuable host for ticks, which is an example of ecological interference. Therefore, Mycoplasma has the potential to ultimately reduce transmission outcomes of tick-borne pathogens via songbird hosts.

Complex interactions between bacteria and haemosporidia in coinfected hosts: An experiment.

Hosts are typically coinfected by multiple parasite species whose interactions might be synergetic or antagonistic, producing unpredictable physiological and pathological impacts on the host. This study shows the interaction between Plasmodium spp. and Leucocytozoon spp. in birds experimentally infected or not infected with Mycoplasma gallisepticum.In 1994, the bacterium Mycoplasma gallisepticum jumped from poultry to wild birds in which it caused a major epidemic in North America. Birds infected with M. gallisepticum show conjunctivitis as well as increased levels of corticosterone.Malaria and other haemosporidia are widespread in birds, and chronic infections become apparent with the detectable presence of the parasite in peripheral blood in response to elevated levels of natural or experimental corticosterone levels.Knowing the immunosuppressive effect of corticosterone on the avian immune system, we tested the hypothesis that chronic infections of Plasmodium spp. and Leucocytozoon spp. in house finches would respond to experimental inoculation with M. gallisepticum as corticosterone levels are known to increase following inoculation. Plasmodium spp. infection intensity increased within days of M. gallisepticum inoculation as shown both by the appearance of infected erythrocytes and by the increase in the number and the intensity of positive PCR tests. Leucocytozoon spp. infection intensity increased when Plasmodium spp. infection intensity increased, but not in response to M. gallisepticum inoculation. Leucocytozoon spp. and Plasmodium spp. seemed to compete in the host as shown by a negative correlation between the changes in their PCR score when both pathogens were present in the same individual.Host responses to coinfection with multiple pathogens measured by the hematocrit and white blood cell count depended on the haemosporidian community composition. Host investment in the leukocyte response was higher in the single-haemosporidia-infected groups when birds were infected with M. gallisepticum.A trade-off was observed between the immune control of the chronic infection (Plasmodium spp./Leucocytozoon spp.) and the immune response to the novel bacterial infection (M. gallisepticum).

Haemosporidian parasite prevalence, parasitemia, and diversity in three resident bird species at a shrubland dominated landscape of the Mexican highland plateau.

BACKGROUND: Studies of avian haemosporidians allow understanding how these parasites affect wild bird populations, and if their presence is related to factors such as habitat loss, degradation and fragmentation, and climate change. Considering the importance of the highland Plateau of Mexico as part of the North American bird migratory route and as a region containing important habitat for numerous bird species, the purpose of this study was to document haemosporidian species richness and how habitat degradation, bird body condition, and distance from water sources correlate with bird parasitemia. METHODS: We assessed the presence of avian haemosporidians in three resident bird species through microscopy and PCR amplification of a fragment of the haemosporidian cytochrome b gene. Average parasitemia was estimated in each species, and its relationship with habitat degradation through grazing, bird body condition and distance from water bodies was assessed. RESULTS: High levels of parasitemia were recorded in two of the three bird species included in this study. Four lineages of haemosporidians were identified in the study area with nearly 50 % prevalence. Areas with highly degraded shrublands and villages showed higher parasitemia relative to areas with moderately degraded shrublands. No strong relationship between parasitemia and distance from water bodies was observed. There were no significant differences in prevalence and parasitemia between the two bird species infected with the parasites. Two of the sequences obtained from the fragments of the parasite's cytochrome b gene represent a lineage that had not been previously reported. CONCLUSIONS: Haemosporidian diversity in arid zones of the Mexican highland plateau is high. Shrubland habitat degradation associated to the establishment of small villages, as well as tree extraction and overgrazing in the surroundings of these villages, significantly enhances parasitemia of birds by haemosporidians.