Vol.:(0123456789) 1 3 Acta Parasitologica https://doi.org/10.2478/s11686-019-00123-y ORIGINAL PAPER Comparison of Light Microscopy and Polymerase Chain Reaction for the Detection of Haemoparasites in Cattle in Nigeria Anise N. Happi 1  · Olawale Osifade 1  · Paul E. Oluniyi 2,3  · Bamidele N. Ogunro 4 Received: 23 May 2019 / Accepted: 14 September 2019 © Witold Stefański Institute of Parasitology, Polish Academy of Sciences 2019 Abstract Purpose Haemoparasitic diseases are among the important factors that threaten cattle health and productivity especially in the sub-Saharan region. In Nigeria, their detection using sensitive molecular techniques is scanty. This study was designed to investigate and to reevaluate the repertoire of haemoparasites of cattle in Ibadan, Nigeria with a comparative evaluation of light microscopy (LM) and polymerase chain reaction (PCR) methods. Methods Blood samples from 100 cattle slaughtered at Ibadan abattoirs were examined using LM and PCR techniques for haemoparasite detection. The PCR reactions using three primer sets targeting the 16S rRNA genes for Hemoplasma spp. and Anaplasma/Ehrlichia spp. and 18S rRNA genes of Babesia/Theleiria spp. were done. A few randomly selected amplicons from each set were sequenced and analysed. Results A total infection rate of 34% by LM including Hemoplasma spp. (17%), Anaplasma spp. (16%), microflaria (5%) and Trypanosoma spp. (12%) was recorded. While, 86% positivity was recorded with PCR amplifcation as follows: Hemo- plasma spp. (64%), Babesia/Theleiria spp. (46%) and Anaplasma/Ehrlichia spp. (5%). Comparison of LM and PCR fndings showed that no LM Anaplasma spp.-positive samples and 7 out of the 17 LM hemoplasma-positive cattle were confrmed by PCR. In addition, LM led to misdiagnosis of 46 Babesia/Theleiria spp.-positive samples. Amplicon sequencing and phylogenetic analysis of Babesia/Theileria spp.-positive samples revealed Theileria velifera and Theileria annulata. In the Anaplasma/Ehrlichia spp.-positive samples, only Anaplasma marginale was characterized. Mycoplasma wenyonii, “Candi- datus Mycoplasma haemobos” and Pseudomonas fluorescens like were characterized among the hemoplasma-infected cattle. Conclusions The frst report of “Candidatus Mycoplasma haemobos” and Pseudomonas fluorescens like in Nigerian cattle is herewith documented. The alarming LM misdiagnosis of haemoparasites during this study confrms its limitations as it fails to identify many parasites and emphasizes the need for inclusion of molecular techniques to improve their detection. The study also shows for the frst time the high prevalence of haemotropic mycoplasma in Nigerian cattle via molecular diagnostic methods, thus indicating a strong need for the investigation of their zoonotic implications. Keywords Cattle · Nigeria · Haemoparasites · Mycoplasma haemobos · Pseudomonas fluorescens like Introduction Cattle production is still signifcantly low in the tropical and subtropical regions due to many factors among which are malnutrition, poor management, poor genetic potential as well as diseases. Haemoparasitic diseases rank relatively high among factors that threaten cattle productivity espe- cially in the tropical and subtropical regions [1, 2]. They have generally been shown to cause destruction of red blood cells resulting in anaemia, jaundice, anorexia, weight loss, decrease in milk production, infertility and various tissue pathology. Some of these haemoparasitic diseases also have debilitating impact on animal and human health worldwide particularly in developing countries [3]. Their diagnoses in most resource-limited areas, however, remain mainly limited to light microscopy (LM). Haemoparasites can be diagnosed by identifcation of the parasites in blood or tissues (lymph node, liver, spleen, brain, kidney, heart muscle) by LM, polymerase chain reaction (PCR), serology or animal inoculation experi- ments [4]. In Nigeria, microscopic diagnosis remains the most common and routinely used diagnostic method for * Anise N. Happi anisehappi@yahoo.com Extended author information available on the last page of the article