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Haemoproteus spp. and Leucocytozoon californicus Coinfection in a Merlin (Falco colombarius).

ABSTRACT: The Leucocytozoon genus comprises numerous widely distributed parasites which have been less investigated than other avian hemoprotozoa. Their occurrence is common, with very variable prevalence values and pathogenicity degrees. Leucocytozoon species are characterized by a great taxonomic diversity, and infections are usually restricted to birds of the same family. In the present paper, a mixed hemosporidia infection by Leucocytozoon californicus and Haemoproteus sp. in an adult male merlin (Falco columbarius) which died during hospitalisation is reported, indicating, for the first time, a newly described avian host species. A molecular investigation was carried out through cytochrome b gene analysis, revealing a 100% match with L. californicus and Haemoproteus spp. A blood smear examination allowed us to detect Leucocytozoon fusiform mature gametocytes and different degrees of maturity of Haemoproteus gametocytes. Histopathology revealed foci of necrosis, hemorrhagic areas and extramedullary hematopoiesis in the liver, the presence of microthrombi in the heart and lung and scattered hemorrhages in the lung.

SUBMITTER: Nardoni S

PROVIDER: S-EPMC7238121 | biostudies-literature | 2020 Apr

REPOSITORIES: biostudies-literature

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Haemoproteus spp. and Leucocytozoon californicus Coinfection in a Merlin (Falco colombarius).

Nardoni Simona S Parisi Francesca F Rocchigiani Guido G Ceccherelli Renato R Mancianti Francesca F Poli Alessandro A

Pathogens (Basel, Switzerland) 20200404 4

The Leucocytozoon genus comprises numerous widely distributed parasites which have been less investigated than other avian hemoprotozoa. Their occurrence is common, with very variable prevalence values and pathogenicity degrees. Leucocytozoon species are characterized by a great taxonomic diversity, and infections are usually restricted to birds of the same family. In the present paper, a mixed hemosporidia infection by Leucocytozoon californicus and Haemoproteus sp. ...[more]

PMID: 32260378

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Project description:BackgroundAvian haemosporidian parasites can cause severe disease in their hosts due to excessive exo-erythrocytic merogony and anaemia caused by blood stages. Notably, the development of megalomeronts by species of Haemoproteus and Leucocytozoon has been associated with mortalities in birds. Diagnosis of lethal infections is currently accomplished by the detection of parasites' tissue stages in histological sections combined with PCR and sequencing. However, sequences frequently are not reliably obtained and the generic discrimination of exo-erythrocytic tissue stages based on morphological characters is challenging. Therefore, the present study aimed at developing specific molecular probes for the identification of Haemoproteus spp. and Leucocytozoon spp. in histological sections using chromogenic in situ hybridization.MethodsParasite subgenus-specific oligonucleotide probes were designed to target the 18S ribosomal RNA of Haemoproteus species (subgenus Parahaemoproteus) and Leucocytozoon spp. (subgenus Leucocytozoon) and were in situ hybridized to sections from formalin-fixed, paraffin-embedded tissue samples determined positive for these parasites by PCR and histopathology. To confirm the presence of parasites at sites of probe hybridization, consecutive sections were stained with haematoxylin-eosin and examined.ResultsParahaemoproteus- and Leucocytozoon-specific probes labelled erythrocytic and exo-erythrocytic stages of Haemoproteus spp. and Leucocytozoon spp., respectively. Binding of probes to parasites was consistent with detection of the same exo-erythrocytic meronts in consecutive haematoxylin-eosin-stained sections. Cross-reactivity of the probes was ruled out by negative chromogenic in situ hybridization when applied to samples positive for a parasite of a genus different from the probes' target.ConclusionsChromogenic in situ hybridization using 18S ribosomal RNA-specific oligonucleotide probes reliably identifies and discriminates Haemoproteus and Leucocytozoon parasites in tissue sections and enables unequivocal diagnosis of haemosporidioses.

Project description:During a 6-year period, 64 of 227 commercially reared cats had microaerobic bacteria isolated from their feces. All the isolates were initially identified as Campylobacter-like organisms based on biochemical and phenotypic characteristics. DNA extractions from 51 of these isolates were subjected to PCR using primers specific for Helicobacter spp. and Campylobacter spp. Of the isolates, 92% (47 of 51 isolates) were positive for Campylobacter spp., 41% (21 of 51 isolates) were positive for Helicobacter spp., 33% (17 of 51 isolates) were positive for both genera, 59% (30 of 51 isolates) were positive only for Campylobacter spp., and 8% (4 of 51) were positive only for Helicobacter spp. Sixteen of the 47 Campylobacter-positive cultures were positive for more than one Campylobacter spp. Based on a species-specific PCR assay, 83% of the isolates were identified as Campylobacter helveticus, 47% of the isolates were identified as Campylobacter upsaliensis, and 6% of the isolates were classified as Campylobacter jejuni. The 1.2-kb PCR products of the 16S rRNA genes of 19 Helicobacter species isolates were subjected to restriction fragment length polymorphism (RFLP) analysis. Of the five different RFLP patterns obtained, two clustered with Helicobacter ("Flexispira") taxon 8, one clustered with Helicobacter bilis, one clustered with Helicobacter canis, and the remaining pattern was closely related to a novel Helicobacter sp. strain isolated from a woodchuck. The sequence data for the 16S rRNA genes of 10 Helicobacter spp. validated the RFLP-based identification of these isolates. This study demonstrated that biochemical and phenotypic characteristics of microaerobic organisms in cat feces were insufficient to characterize mixed Helicobacter and Campylobacter infections. Molecular structure-based diagnostics using genus- and species-specific PCR, RFLP analysis, and 16S rRNA sequence analysis enabled the identification of multiple microaerobic species in individual animals. The clinical relevance of enteric Helicobacter and Campylobacter coinfection in cats will require further studies.

Project description:BackgroundA high prevalence of parasites may result from life-long persistence of infection or from high reinfection rates. We have studied blood parasites in a breeding population of the accipitrid raptor, Eurasian sparrowhawk (Accipiter nisus), to determine parasite diversity and turnover.MethodsDuring this 7-year study, 210 adult Eurasian sparrowhawks breeding in the city of Prague were checked for parasites using several diagnostic methods.ResultsIn both female and male raptors, parasites of the genus Leucocytozoon were the most prevalent (92% and 85%, respectively) followed in decreasing order of prevalence by those of genus Trypanosoma (74% and 68%, respectively) and genus Haemoproteus (46% and 16%, respectively). The prevalence of all parasites increased with age in both sexes, with the females at each respective age having the higher prevalence. There was a positive association between Haemoproteus and Leucocytozoon infections. Persistence at the individual level was higher than incidence for Trypanosoma and Haemoproteus. In the case of Leucocytozoon and Trypanosoma, most individuals probably become infected in their first year of life or even before dispersal from the nest. The detected parasites belonged to Trypanosoma avium sensu stricto, Leucocytozoon sp. (haplotypes ACNI1 and ACNI3) and Leucocytozoon mathisi (haplotype ACNI4) and two new lineages of the Haemoproteus elani complex (ACCNIS6 and ACCNIS7). Detailed analysis of parasite lineages in individuals that were repeatedly sampled revealed lineage turnover that would otherwise remain hidden. Phylogenetic analysis revealed that the detected Haemoproteus belongs to a phylogenetically distant group whose taxonomic position requires further analysis.ConclusionsAll three genera of blood parasites persist in infected individuals, thus enabling sustainability of vector transmission cycles. Prevalence increases with age; however, there is a high turnover of Leucocytozoon lineages. No clear evidence of parasite-induced mortality was found, and most of the individuals were infected early in life, particularly in the case of Leucocytozoon.

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Haemoproteus spp. and Leucocytozoon californicus Coinfection in a Merlin (Falco colombarius).

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<i>Haemoproteus</i> spp. and <i>Leucocytozoon californicus</i> Coinfection in a Merlin (<i>Falco colombarius</i>).

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