Detection of Haemoparasites of Blood Donors in 9 Locations in and Around Plateau State, Nigeria

Haemoparasites in the tropics are also endemic in Nigeria. Asymptomatic infections may abound, due to resistance to these infections. This asymptomatic infection has been one of the factors, which has maintained transmission of these pathogens, through many ways, including blood donation and transfusion. In this report, haemoparasitic infections in blood donors have been described, from blood donors within Plateau State, Nigeria. Five hundred and twelve blood donors were selected by means of a random sampling method and their blood samples collected. Serological assay was done using rapid test kits to check for presence of antibodies (in the case of microfilariae) or antigens (in the case of malaria) to the different haemoparasites. Also, Elisa technique was used for the microfilariae. Thick and thin films were made from each blood sample on grease-free slides allowed to dry and stained by 3% Giemsa solution for 45 min which is the Giemsa technique. Results indicate that 270 (52.7%) of the sample population had no infection; 121( 23.6%) of the population were infected with Plasmodium falciparum; 11 (2.1%) were infected with Plasmodium malariae; 69 (13.5%) were infected with HBsAg; 29 (5.7%) were infected with HCV; 7 ( I.4% ) were infected with Trypanosoma brucei gambiense; 1% were infected with microfilariae, 4( 0.8% ) of the 1% were unsheathed and identified to be Mansonella perstans, while 1(0.2%) were sheathed and identified to be Loa loa. Most blood group types were susceptible to haemoparasitic infections. The result of the study therefore stresses the need to screen blood for haemoparasites before transfusion, owing to the dangers of doing otherwise. The occupations and dwelling places of the donors are predisposing factors to these haemoparasitic infections. Since they have the passion to save lives through blood donation, they should therefore make the necessary adjustments that will make them more suitable lifesavers. It is recommended that the basic transmission factors of these parasites are explained to donors to reduce further incidences. DOI: 10.7176/JBAH/9-22-01 Publication date: November 3


Introduction
Blood transfusion is considered a life-saving intervention that has an essential role in patient management within health care systems. All Member States of the World Health Organization (WHO) endorsed World Health Assembly resolutions [1] and [2] in 2005.
The establishment of systems, (like National Blood Transfusion Service in Nigeria), that ensure that all donated blood is screened for transfusion-transmissible infections is a core component of every national blood programme. Globally, however, there are significant variations in the extent to which donated blood is screened, the screening strategies adopted and the overall quality and effectiveness of the blood screening process. As a result, in many countries the recipients of blood and blood products remain at unacceptable risk of acquiring lifethreatening infections that could easily be prevented [2,3].
The range of investigations carried out for donors need to be increased in practice, since transfusion reactions still occur despite complete cross-match of donor cells and recipient serum. This suggests the presence of some antigens which recipient develops antibodies to, hence a reaction. Not all reactions are therefore elicited as a result of incompatibility of the blood being given [4]. Haemoparasite infections are some of the commonest and most important diseases (antigens) of man in the tropics and subtropics. These infections are an important constraint to man since it affects man socially, economically and otherwise [5].
The aim of this study is to detect haemoparasites of among blood donors recruited from 9 locations in and around plateau state, Nigeria.

Study Area/Sample Size
The study was carried out in Jos, Plateau state and environs. Five hundred and twelve donors were selected by means of a random sampling method and their blood samples collected. Analyses were made for haemoparasites.
The blood groups was also determined and recorded.
Prior to sample collection, an ethical clearance was obtained from NBTS ethical committee and oral consent was obtained from the donors before their names were inputted into the detail collection form.

Sample Collection
Blood samples were collected using sterile EDTA. This was achieved by tying a tourniquet above a prominent vein, and a sterile swab used to clean the area to reduce the risk of contamination. With a sterile syringe, blood is collected from the vein and immediately put into an EDTA bottle. The blood was mixed gently to avoid coagulation. All samples collected were properly labeled, kept in ice and transported to the Angel of Hope Clinic or DEE Medical Centre laboratory for processing within 3 hours of collection.

Processing of Samples
Serological assay was done using rapid test kits to check for presence of antibodies to (in the case of other haemoparasites) or antigens of (in the case of malaria parasites) the different haemoparasites. The various tests were run following manufacturer's guidelines/instructions. Microfilaria was screened for using ELISA -Og4C3 monoclonal antibody assay, which is quantitative. The Leishmania and Trypanosoma serology kit was not readily available for this research. Tests are positive for the particular blood parasite analyzed, if a double band was seen (for the rapid test kits) and agglutination or adhesion (for the ELISA). It was however confirmed using other test methods like microscopy, which has remained a standard for malaria and diagnosis of other haemoparasites [6]. Parasitological examination: Thick and thin films were made in duplicates from each blood sample on greasefree slides, allowed to dry and stained by 3% Giemsa solution for 45 min which is the Geimsa technique [6]. Stained blood films were examined under x100 objective lens of a microscope with the aid of immersion oil for haemoparasites. Thin films were used for the species identification of parasites. Further confirmation of positive samples were undertaken to ensure quality control. Wet preparation and Delafield's haemalum Staining technique for microfilaria: This stain is a derivative of Haematoxylin and alum. It stains the nuclear matter. Eosin is used with it as a counter stain. The staining procedure was carried out according to the methods of Cheesbrough [6]. Determination of ABO blood group: Three spots of blood from each subject were made on the white plain tile and a drop of each antiserum A, B and D was applied to each of the different spots of blood, respectively. The mixture was further stirred with a plastic stirrer and rocked for 2 minutes. A sign of agglutination on any of the spots was observed and recorded [6].

Statistical Analysis
Data obtained were subjected to statistical analyses (using mean of occurrence, percentage prevalence and Non Parametric Chi-Square test).  This table indicates the parasite spectrum in relation to blood group types. This is however a bit skewed and not holistic, since all blood group types was not available for the donation.

Results
For those that however donated, Blood group O positive was highest for all parasite types. This was followed by Blood group B positive, then Blood group A positive. There were however cases where Blood group A positive recorded more infection than Blood group B positive (reference). It is however important to note that no matter the blood group, parasitaemia is possible. This means that every blood group is susceptible to blood parasitic infections.  The parasite intensity among the donors is recorded in table 1. The most intense parasitic infection is P. falciparum with the highest incidence and L. loa is the least intense infection.  Table 3 shows the haemoparasitic infection in relation to the occupation of the donors. The total number of each occupation type that had any form of haemoparasitic infection was compared with the total number of donors that were encountered in that occupation type. This relationship, showed housewives to be the highest susceptible group and artisans the lowest.  Table 4 shows the haemoparasitic infection in relation to the location of the donors recruited. The total number The work also showed the spread of the haemoparasitic infection depending on the occupation of the donors and the various locations of donor recruitment. This is to be investigated further, for a more meaningful discussion in these areas. It can however be said that more rural settlements had more infection than the more urban settlements. Predisposing factors will be further investigated. Also, the elites had lesser infection than the housewives and farmers. Also, this is an opening for further investigation.

Conclusion
This study reveals that the infection rate of the haemoparasites differs, as well as parasite intensity. The highest infection occurred due to P. falciparum, with an infection rate of as high as 23.6% while the lowest infection was due to L. loa, with an infection rate of 0.2%. It was observed that parasitaemia was possible, regardless of the blood group type.
This study therefore concludes and stresses the need to take parasites associated with blood transfusion very seriously, considering the dangers of doing otherwise. There could be evidence and a possibility of evolutionary mechanisms of attenuation of symptoms of haemoparasitic infections in humans.
Also it is important to note that the people in need of blood transfusion in very many cases may not have all it takes to withstand invasion of these parasite. With this in mind, it is expedient to ensure that the necessary precautions are taken to avoid fatal consequences. Therefore, This study contribute to knowledge in the following ways; according to the criteria given by World health Organization in 2010 (16) Plasmodium falciparum, Plasmodium malariae, Trypanosoma brucei gambiense, Mansonella perstans (unsheathed microfilaria), loa loa (sheathed microfilaria) were not included in the mandatory minimum requirement for screening of blood donor for infection. With our findings, it is necessary that it should be included because of Current and emerging infectious risks of blood transfusion (17,18)..

Recommendations
Following the findings, the following recommendations are tendered: 1. Proper screening of blood for all haemoparasites before blood is transfused to recipient should be carried out.
2. Recipients may be asymptomatic after contracting a post transfusion infection, hence carriers may constitute a reservoir for parasites, further maintaining transmission. This calls for the need for an after transfusion screening and documentation of the risk of transfusion related haemoparasitic infections.

Conflicts of Interest
There is no conflict of interest.