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I. Schäfer, M. Faucher, Y. Nachum-Biala, L. Ferrer, M. Carrasco, A. Kehl, E. Müller, T.J. Naucke, G. Baneth

Parasites & Vectors volume 17, Article number: 506 (2024)

Abstract

Background: In Europe, canine leishmaniasis is commonly caused by Leishmania infantum. Allopurinol is the main drug for long-term management of the disease, and clinical relapses of L. infantum infection treated with this drug are described. Resistance to allopurinol has been demonstrated in-vitro, but there is only little knowledge on in vivo resistance in dogs.

Findings: A two-year-old female spayed Akita Inu that was adopted from a breeding facility near Nice in France was initially diagnosed with primary immune-mediated hemolytic anemia. Immunosuppressive treatment was initiated, and the dog was referred for a second opinion to the Clinique Veterinaire Alliance in France. PCR testing for L. infantum was performed out of EDTA blood and IFA as well as ELISA testing out of serum. Resistance to allopurinol was associated with chromosome and gene copy number (CN) variations including a decrease in the S-adenosylmethionine synthetase (METK) gene CN.Results

Results: The dog showed pale mucous membranes, fever (39.1 °C), and a relapse of the anemia. The diagnosis of leishmaniasis was based on the cytological finding of Leishmania amastigotes (bone marrow, spleen, liver), positive PCR testing, and positive IFAT serology. The dog was treated with allopurinol over a period of 1316 days and additionally received two cycles of Glucantime® (meglumine antimoniate), before samples were submitted to the LABOKLIN laboratory to test for resistance against allopurinol. The laboratory work-up revealed mild thrombocytopenia, mild hyperproteinemia with hyperglobulinemia, a marked elevation of the c-reactive protein, and decreased iron concentration. Serum protein electrophoresis showed a polyclonal peak in the gamma globulins. Serology was positive in both ELISA (21.5 LE) and IFAT (1:1024). Quantitative PCR testing of blood was positive with low numbers of Leishmania (10/ml blood) at the timepoint of suspicion for resistance. The urinary protein-to-creatinine ratio was markedly elevated (2.5) and xanthine crystalluria was detected. A CN level of below 3 is considered suspicious for resistance, as revealed in the described Akita Inu dog.

Conclusions: Relapse of L. infantum infection after applying allopurinol for 1316 days due to resistance was suspected clinically. Positive PCR testing, consistent hematological and biochemistry abnormalities, and reduction in the METK gene CN backed up the clinical suspicion of resistance. Dogs infected with allopurinol resistant strains of L. infantum may represent a great risk for infection of naïve dogs, cats, and humans.

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C.W. Axt, A. Springer, C. Strube, C. Jung, T.J. Naucke, E. Müller, I. Schäfer

Microorganisms 2024, 12(4), 816

Abstract: Equine piroplasmosis (EP) is caused by Theileria (T.) equi and/or Babesia (B.) caballi. The aim was to assess the percentage of positive test results for EP in horses in Europe and to identify risk factors for pathogen contact/infection. This study included results from PCR and competitive enzyme-linked immunosorbent assay testing requested by European veterinarians between 2008 and 2021. Binary bivariate logistic regression was used to analyze risk factors. A total of 4060 horses were included. PCR testing was positive in 9.7% (154/1589), serology for T. equi in 15.2% (393/2591) and for B. caballi in 6.8% (175/2578). The odds of positive serology increased by 6.8% (B. caballi, p = 0.008) and 9.5% (T. equi, p < 0.001) each year. Regionality had a statistically significant impact on PCR (Eastern p = 0.047/OR = 1.605; Southern p = 0.029/OR = 1.451; Central p = 0.007/OR = 0.617) and serological testing for T. equi (Southern p < 0.001/OR = 2.521; Central p < 0.001/OR = 0.537; Northern p = 0.003/OR = 0.462), as well as breeds on seroprevalence of B. caballi (heavy horses: p = 0.016/OR = 2.239) and T. equi (ponies: p = 0.007/OR = 0.340; warmbloods: p = 0.025/OR = 1.602). In conclusion, there was a significant geographical impact on the results of PCR and serology, consistent with known vector habitats. The rising numbers of horses tested serologically positive highlights the importance of surveillance.

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Zusammenfassung: Aufgrund von steigenden Importzahlen von Hunden aus dem Ausland, zunehmendem Reiseverkehr sowie den Veränderungen klimatischer Bedingungen in Europa gewinnen Infektionen mit Leishmania (L.) infantum bei Hunden in Deutschland zunehmend an Bedeutung. Daher sollten auch Hunde aus dem Ausland, die keine klinischen Symptome zeigen, direkt nach Import sowie erneut 6 Monate später auf vektorübertragene Infektionserreger getestet werden. Bei Hunden mit klinischer Symptomatik, die hinweisend auf eine Leishmaniose sind, werden direkte und indirekte Nachweisverfahren sowie eine hämatologische und biochemische Untersuchung unter Einbezug von Serumeiweißelektrophorese sowie Bestimmung des C-reaktiven Proteins empfohlen. Als Leitfaden für die Therapie sowie das Monitoring stehen die LeishVet-Guidelines zur Verfügung. Es stehen leishmanizide und leishmaniostatische Wirkstoffe zur Verfügung, die in first-line, second-line und third-line unterschieden werden. Zur Anpassung der Allopurinol-Dosierung wird der Stufenplan empfohlen. Aufgrund der Veränderung der klimatischen Bedingungen kommt es zu einer Ausbreitung der Habitate von Sandmücken, die als Vektoren der Leishmaniose bekannt sind. Als weitere Infektionsquellen sind Deckakte, transplazentare Infektionen, Bisswunden und Bluttransfusionen beschrieben. Leishmania infantum hat zoonotisches Potential und ist daher auch in Hinblick auf den „One-Health“-Gedanken bedeutend.  

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I. Schäfer, E. Müller, A.M. Nijhof, H. Aupperle-Lellbach, G. Loesenbeck, S. Cramer, T.J. Naucke

Parasites & Vectors volume 15, Article number: 296 (2022)

Abstract

Background: Hepatozoon canis is a protozoal agent that is known to be transmitted by oral uptake of H. canis‑infected Rhipicephalus sanguineus sensu lato ticks in dogs. Vertical transmission of H. canis has only been describedonce in a study evaluating dogs from Japan. The aim of this study was to investigate the parasitological status of pup‑pies from a bitch that had tested positive for Hepatozoon spp. prior to giving birth.

Findings: A 4‑year‑old, female, pregnant dog imported from Italy (Sardinia) to Germany showed clinical signs of leth‑argy and tachypnoea and tested positive for H. canis by PCR. The dog gave birth to eight puppies, one of which wasstillborn and another that had to be reanimated. Haematology, buffy coat analysis and a biochemistry profile wereperformed for each dog. EDTA‑blood of the surviving seven puppies and bone marrow, liver, spleen, amniotic fluid,and umbilical cord of the stillborn puppy was tested for the presence of Hepatozoon spp. by PCR. The mother and the seven surviving puppies tested positive for H. canis by PCR at day 62 post‑partum. Gamonts weredetected in all dogs by buffy coat evaluation. Haematological and biochemistry results revealed mild abnormalities. Inthe stillborn puppy, spleen, umbilical cord, and amniotic fluid were positive for H. canis.

Conclusion: The results confirm that vertical transmission is a possible route of H. canis infection in dogs, dem‑onstrated by molecular detection of the pathogen in the stillborn puppy. In the seven surviving puppies, vertical transmission was the most likely transmission route. A potential impact of the level of parasitaemia on the health of puppies, as well as its pathogenesis, should be investigated further.

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