Genetic characterization of Trichomonas gallinae (Rivolta, 1878) in companion birds in Japan and the genotypical relationship in the Asia region.

BACKGROUND/PURPOSE: Avian trichomonosis is a parasitic infection that affects a wide range of avian species, including free-ranging and pet birds worldwide, and Trichomonas gallinae has been considered as the only causative agent for decades. The sequence of the 5.8S ribosomal RNA with internal transcribed spacer (ITS) regions was widely used for identifying genotypes and determining inter-specific and intra-specific diversity. Moreover, the sequence of Fe-hydrogenase (FeHyd) was proposed as the second genetic marker for providing improved resolution of strain subtyping discrimination. Though the correlation between genetic variability and strain virulence is controversial, FeHyd analyses seemed to be useful to investigate the host or geographic origin of isolates. This study aimed to investigate the genetic characteristics of avian Trichomonas spp. METHODS: Forty-seven oral swabs and crop lavage fluids were collected from 9 avian genera, which were diagnosed as Trichomonas-positive by microscopy in animal hospitals in Japan, were analyzed. RESULTS: Genetic analysis of clonal isolates revealed the prevalence of the single genotype, ITS-OBT-Tg-1, by ITS region analysis, while two different subtypes, A2 and novel A3, were suggested by FeHyd gene analysis among Japanese companion birds. Phylogenetic analyses of available ITS sequences obtained from the Asia region (China, Iran, Iraq, and Saudi Arabia) were also preformed, revealing endemic ITS-OBT-Tg-1, ITS-OBT-Tg-2, ITS-OBT-Ttl-1, genotype III, and Saudi Arabia's unique lineages. Furthermore, ITS-OBT-Tg-2 predominance in these countries indicates different strains origination from Japan. CONCLUSION: This is the first report of the genetic characterization of T. gallinae in Japan with discovery of novel subtype A3.

Eosinophilic Leukaemia and Systemic Mycobacterium marinum Infection in an African Pygmy Hedgehog (Atelerix albiventris).

A two-year-old male African pygmy hedgehog (Atelerix albiventris) presented with lethargy and anorexia. Ultrasonographic and radiographic examinations revealed splenomegaly and pneumonia, respectively. Prominent leucocytosis, consisting mainly of large atypical eosinophils, was observed in a peripheral blood smear. Necropsy revealed a black scab on the left hindlimb, which was swollen, an enlarged left inguinal lymph node, firm oedematous lungs, splenomegaly and multiple nodules in the lung, spleen and liver. Histopathologically, infiltration of numerous eosinophils was seen in lung, spleen, liver, kidney, lymph node and adrenal medulla tissues. Necrotizing granulomatous lesions, with intralesional acid-fast bacilli, were found in the lung, spleen, liver, left inguinal lymph node and left hindlimb. Mycobacterium marinum was identified by broad-range polymerase chain reaction targeting of the bacterial 16S rRNA gene. To the best of our knowledge, this is only the second reported case of systemic M. marinum infection in any mammalian species.

Determination of estrogens and their conjugates in water using solid-phase extraction followed by liquid chromatography-tandem mass spectrometry.

An analytical procedure for the determination of steroid estrogens and their conjugates was developed and applied to aqueous environmental samples. The analytes of 15 compounds were solid-phase extracted and fractionated into two fractions: one containing unconjugated (free) steroids and the other containing conjugates (sulfates and glucuronides). Identification and quantification were carried out using liquid chromatography coupled with tandem mass spectrometry. The recoveries for each compound ranged from 57 to 116% and reproducibilities represented as RSD ranged from 2.9 to 17%. Some of the sulfates and free compounds were detected in environmental samples, whereas most of the conjugates were below the detection limits.