PREVALENCE AND HAEMATOLOGICAL INDICES OF GARDIASIS AND MALARIA AMONGST PUPILS OF PAEDIATRIC AGE (0-15) IN OWERRI WEST IMO STATE, NIGERIA

This study investigated the prevalence and haematological parameters of gardiasis and malaria amongst pupils of paediatric age (0-15)in 20 randomly selected primary schools was carried out in Owerri West Local Government Area of Imo State, Nigeria. Observations were made of the age, sex, socio-economic status of parents and the schools locations to determine the secondary outcomes measures to these two diseases. Both blood and faecal samples were collected from each of the pupils between the hours of 8.00 am and 11.00 am using 19cc needle guage and 10 ml syringes into EDTA bottles for the blood and sterile applicator sticks employed for the faeces, into wide-mouthed screwed, non-greasy universal bottles. Exclusion procedures were also carried out to eliminate other possible protozoal parasites including administration of combantrin tablets to the 150 volunteers for deworming. Blood parasitaemia due to malaria parasites was determined using the giemsa stained thick and thin smears, while giardiasis was determined using the faecal wet preparation technique involving the Lugols iodine faecal smears for the presence of trophozoites, cysts or casts. Determinations of other haematological parameters were carried out using histochemical techniques. Results showed out of 150 samples, 78(52.0%) were infected with either giardiasis, 20(25.64%) or other malaria parasite forms 30(38.46%), while 28(35.89%) had mixed infections of both malaria and giardiasis. A total parasite count for both infections was 1958 represented as 37% and 62.92% for giardiasis and malaria respectively. Forty-two (42) samples were discarded due to indeterminate results. RBC counts (X1012/L) for control: scarification (5.6±2.0), unscarification (2.7±0.3) and for test (infected): malaria (4.1±3.2), giardiasis (2.9±1.3), and malaria+giardiasis (1.4±0.02) showed marked significant difference (t=3.7, p<0.002) and similarly between both infection categories (t=1.8, p<0.007). Recorded values for PCV showed control: scarification (42.5±3.3), unscarification (26.2±2.2) and for test (infected): malaria (30.4±4.2), giardiasis (28.4±1.0) are indirectly proportional to intensity of infection establishing significant correlation between parasite count (PC) and PCV (r=0.45, p<0.002), PC and RBC (r=0.36, p<0.002), PC and reticulocyte (r=0.08, p<0.02). MCV values did not show clear correlation pattern with PC (r= -0.07, p>0.02). Mean proteins from the test (infected) groups: total protein (malaria, 6.13±0.01 & giardiasis, 4.50±0.10) albumin – (malaria, 5.52±0.30 & giardiasis, 7.41±2.2) globulin- (malaria, 6.40±0.15 & giardiasis, 8.15±0.22) reduced significantly (p<0.01) compared with each of the control groups and showed significant correlation (r=0.01, p<0.003) with PC: malaria (104.4±3.3x105), and giardiasis (76.2±5.8x105).Giardia lamblia and malaria parasite species are naturally prevalent protozoan parasites. Notable significant difference in prevalence of these protozoan parasites infestations among children abound.

Effect of mast cell stabilization on angiogenesis in primary and secondary experimental Trichinella spiralis infection

Background Mast cells are known to affect the primary and secondary immune responses against parasites, and this effect is partially mediated through the release of pro-angiogenic mediators. The aim of this study was to explore the effect of the mast cell stabilizer (MCS), ketotifen, with and without albendazole, an anti-parasitic prescription medicine, on the inflammatory response against Trichinella spiralis, with the overall aim to investigate its effect on angiogenesis accompanying nurse cell formation. Methods The effect of ketotifen and albendazole was explored in eight groups of female BALB/c mice. Four groups were sensitized with a small dose of T. spiralis larvae. The drug regimen was then applied to both sensitized (challenged) and non-sensitized mice. The parasite load was assessed by histopathological examination of the small intestine and muscle tissue, and angiogenesis was assessed by immunohistochemistry to determine the expression of vascular endothelial growth factor (VEGF). Results Sensitized mice showed a significantly lower parasite load and a more pronounced inflammatory response than mice receiving a single infective dose of T. spiralis larvae. All treated groups showed a significant reduction in parasite count compared to the control groups (groups IAa and IBa), reaching approximately an 98.8% reduction in adult parasite count in the sensitized group treated with albendazole (groups IIAb and IIBb). MCS significantly decreased the parasite count during both the intestinal or muscular phases, reduced tissue inflammation, and decreased local VEGF expression, both in the non-sensitized and sensitized groups. Conclusion Sensitization with a low dose of T. spiralis larvae was found to confer a partial protective immunity against re-infection and to positively affect the study outcomes, thus underlining the importance of vaccination, but after extensive studies. The anti-angiogenic effect of MCS protects against larval encystation during the muscle phase. The anti-angiogenic potential of albendazole suggests that the action of this anti-helminthic during trichinellosis is not confined to structural damage to the parasite cuticle but includes an effect on host immunopathological response. Graphical Abstract

Malaria Severity Score in Malaria Patients Admitted in Critical Care Wards

Introduction: Prognostic scoring system in Intensive Care Unit (ICU) can be generic, which can be applied to any critical illness for which patients are admitted in critical wards or can be disease specific. Malaria Severity Score (MSS) is a disease specific prognostic scoring system. Aim: To study the role of MSS in patients having malaria who were critically ill having multi organ dysfunction and to correlate the score with risk of mortality. Materials and Methods: This longitudinal study was conducted at the Department of General Medicine, SBKSMI & RC, Sumandeep Vidyapeeth, Vadodara, Gujarat, India. Adult patients (>18 years) with falciparum as well as vivax malaria, who had positive peripheral smear malaria and were admitted in ICU/Casualty (Emergency) ward, were taken in the study. The score was calculated on day of admission, day 2 and day 7. The score was analysed between two groups: survivors and non-survivors. Appropriate statistical tests were applied (z-test for two population proportion and Chi-square test for categorical values). The p-value <0.05 was considered as significant. Results: Out of 60 patients, 41 survived and 19 died due to malaria. Mean age of survivors was 38.56±2.27 and of non-survivors 40.21±5.6 years (p=0.718). There were 27 patients of P.vivax, 30 of P. falciparum and three patients of mixed infection; mortality was in 09, 08, 02 patients, respectively. On admission, out of total 60 patients, 10 (16.67%) had 1+, 20 (33.33%) had 2+, 24 (40%) had 3+ and 6 (10%) had 4+ parasite count. There were no patients in 1+ parasite count group, two (10%) in 2+, eleven (45.8%) in 3+ and six (100%) in 4+ parasite count group. Mean MSS was not significantly different on day 0 and day 2 but was higher on day 7 in non-survivor group than in survivors group (p=0.005). Mortality prediction score cut-off was ≥9, which was obtained by plotting Receiver Operating Characteristic (ROC) curve. Mean MSS in non-survivor group was 7.37 on day 0, 6.58 on day 2 and 9.11 on day 7. Thus, MSS score of day 7 gave `prediction reaching cut-off value of ≥9. Conclusion: MSS was found to be a useful prognostic score in severe falciparum/vivax malaria who needs intensive care treatment as sequential score gives significant difference in survivors and non-survivors on seventh day.

Genetic Variation, GWAS and Accuracy of Prediction for Host Resistance to Sparicotyle chrysophrii in Farmed Gilthead Sea Bream (Sparus aurata)

Gilthead sea bream (Sparus aurata) belongs to a group of teleost which has high importance in Mediterranean aquaculture industry. However, industrial production is increasingly compromised by an elevated outbreak of diseases in sea cages, especially a disease caused by monogeneans parasite Sparicotyle chrysophrii. This parasite mainly colonizes gill tissues of host and causes considerable economical losses with mortality and reduction in growth. The aim of current study was to explore the genetics of host resistance against S. chrysophrii and investigate the potential for genomic selection to possibly accelerate genetic progress. To achieve the desired goals, a test population derived from the breeding nucleus of Andromeda Group was produced. This experimental population was established by crossing of parents mated in partial factorial crosses of ∼8 × 8 using 58 sires and 62 dams. The progeny obtained from this mating design was challenged with S. chrysophrii using a controllable cohabitation infection model. At the end of the challenge, fish were recorded for parasite count, and all the recorded fish were tissue sampled for genotyping by sequencing using 2b-RAD methodology. The initial (before challenge test) and the final body weight (after challenge test) of the fish were also recorded. The results obtained through the analysis of phenotypic records (n = 615) and the genotypic data (n = 841, 724 offspring and 117 parents) revealed that the resistance against this parasite is lowly heritable (h2 = 0.147 with pedigree and 0.137 with genomic information). We observed moderately favorable genetic correlation (Rg = −0.549 to −0.807) between production traits (i.e., body weight and specific growth rate) and parasite count, which signals a possibility of indirect selection. A locus at linkage group 17 was identified that surpassed chromosome-wide Bonferroni threshold which explained 22.68% of the total genetic variance, and might be playing role in producing genetic variation. The accuracy of prediction was improved by 8% with genomic information compared to pedigree.

Inter-rater Variability in Malaria Microscopy at the LEKMA Hospital, Ghana

Background. Malaria remains a major cause of morbidity and mortality worldwide and particularly in sub-Saharan Africa where it is endemic. As such, it is important that a proper diagnosis is made before treatment is initiated. Malaria parasite count plays a key role in the diagnosis and management of malaria. Variations in ratings by laboratory personnel can impact negatively on the treatment regimen for malaria-infected patients. The study is thus aimed at evaluating and comparing the proficiency and parasitaemia counts by two different categories of laboratory staff at the LEKMA Hospital, Ghana. Materials and Methods. A total of 200 confirmed malaria-positive samples were used in the study. Six thick and thin films were prepared from each sample and uniquely labelled. Two of the six slides were given to two WHO-accredited malaria microscopists to examine and report their respective parasite count/μl ( parasite count / WBC × 8000 ). These were used as the reference for the two categories of laboratory staffs: rater A being diploma holders (Technical Officers referred to as untrained rater) and rater B being degree holders (Medical Laboratory Scientist referred to as trained rater) at the LEKMA Hospital. Results. In comparison to the expected outcome, the parasite count by the rater group A (190 (151-239)]) and the rater group B (177 (140-224)) demonstrated significant positive correlation ( r = 0.995 , p < 0.0001 vs. r = 0.995 , p < 0.0001 , respectively) with the expected outcome in the cases of heavy parasitaemia. A statistically significant difference ( p < 0.05 ) between counts by the different raters in low parasitemia was observed in this study. A persistent nosedive inter-rater agreement from k = 0.82 to k = 0.40 with increasing density cutoff was observed in this study. Conclusion. The study observed that the degree of inter-rater agreement of parasite density count by various categories of laboratory personnel is almost perfect. However, the parasite count between raters varied significantly with very low levels of parasitemia but better correlated with heavy parasitemia.

Usefulness of rapid diagnostic test in the diagnosis of asymptomatic malaria in HIV infected children on cotrimoxazole prophylaxis in Benin City, Nigeria

Background: Rapid diagnostic test (mRDT) is a useful tool in demonstrating parasitologically proven malaria. Its efficacy is however hampered when parasite density is low. Prophylactic use of cotrimoxazoleas in cases of HIV infected children can cause reduction in parasite count. It is doubtful if mRDT will retain its diagnostic usefulness among such individuals.Objectives: The study sought to evaluate the diagnostic value of mRDT in HIV infected children on cotrimoxazole prophylaxis in Benin City.Methods: In the prospective, cross sectional and descriptive study, we assessed malaria parasitaemia using standard methods in microscopy and parasite density and malaria antigenaemia using Care Start Pf (monoclonal antibodies specific to histidine rich protein – 2 antigen) in 221 each of HIV infected subjects on cotrimoxazole managed in a specialist clinic and HIV negative controls all seen at the University of Benin Teaching Hospital between April and June 2016.Results: Malaria antigenaemia rate MAr (20.8%) was lower than malaria parasitaemia rate MPr (24.4%) in subjects. MAr (20.8) and MPr (24.4%) in subjects were higher than MAr (18.10%) and MPr (17.7%) in controls. Mean (SEM) parasite count in subjects of was low (50.88 + 2.24 per μl). Using microscopy as gold standard the sensitivity, specificity, PPV and NPV of mRDT in subjects were 77.8%, 97.6%, 91.3% and 93.1%. Corresponding values in controls were 100.0%, 99.5%, 97.5% and 100.0%.Youden indices for subjects and controls were 0.75 and 0.99.Conclusions/Recommendations: Sensitivity of mRDT in HIV infected children on cotrimoxazole prophylaxis for opportunistic infections (OI) is reduced. However the indices of specificity, PPV and NPV are high enough to retain its value in the evaluation of HIV infected children for asymptomatic malaria and perhaps the clinical disease. Keywords: mRDT, Utility, HIVinfected Children, Cotrimoxazoleprophylaxis, Benin City

Impact of Sever Plasmodium falciparum infection on Platelets Parameters among Sudanese children Living in Al-Jazira State

Background: Falciparum malaria remains one of the most global infection among children particularly in communities with poor resources. Falciparum malaria associated with several hematological changes that affect the major blood cell lines such as platelets lead to platelets parameters (platelets count and indices) abnormalities. Objectives: The aim of this study was to evaluate the effects of falciparum malaria on platelets parameters (platelets count and indices) among Sudanese children. In addition to study relationships and correlation between platelets parameters and malaria parasitemia and parasite count. Materials and Methods: A case control study was conducted in Wad Medani Pediatric Hospital in collaboration with Faculty of Medical laboratory Sciences, University of Gezira, Sudan among 100 children with severe falciparum malaria (mean age 8.63 ± 3.40 years; 61% males), 100 children with uncomplicated falciparum malaria (mean age 8.83 ± 4.20 years; 45% males) and 100 children with normal healthy children controls (mean age 10.08 ± 3.58 years; 50% males). Parasitemia and parasite count (%) was determined directly from thick and thin blood films respectively. The platelets parameters (platelets count and indices) measured by using Sysmex XP 300 N automated analyzer, and platelets count was confirmed and assessed using stained thin blood film. SPSS software (V 20.0) and Stat disk software (V 13.0) were used for data analysis. Results: 72 % of severe falciparum malaria (SM) have hyperparasitemia, while 18 % among uncomplicated falciparum malaria (UM). The thrombocytopenia account for 43 % (SM: 30.5 %; UM: 12.5 %), low PCT account for 35.5 % (SM: 27 %; UM: 8.5 %) and high PDW account for 46.5 % (SM: 23.5 %; UM: 23 %) in falciparum malaria cases. The mean PLTs count and PDW were statistically significantly differences between falciparum malaria cases and normal healthy control (P value 0.000 and 0.008 respectively). The mean PLTs count and PCT in severe falciparum malaria cases were lower than uncomplicated falciparum malaria cases (P value 0.005 and 0.000 respectively). The PLTs count and PCT had significant negative correlation within malaria parasitemia (P value 0.000; r -0.286; P value 0.004; r -0.205 respectively) and malaria parasite count (P value 0.000; r -0.450; P value 0.000; r -0.270 respectively). Conclusion: The study concluded that thrombocytopenia, low PCT and high PDW were observed as most platelets parameters changes in falciparum malaria. PLTs count along with PCT to be recommended as hematological diagnostic markers and prognostic tool to assess the disease severity and to improve the management of falciparum malaria among patients.

Molecular Epidemiology of Plasmodium falciparum Chloroquine Resistance Transporter Genes among School Children in Kwara State, Southwestern Nigeria

Background: Plasmodium falciparum existence continues to develop resistance to conventional antimalaria drugs in malaria endemic areas. Plasmodia often prevent drugs from interacting with the target site, hence, developing resistance to antimalaria drugs. Mutations in the Plasmodium falciparum chloroquine resistance transporter (Pfcrt), are the major determinant of chloroquine resistance in human malaria parasite. Methodology: Malaria infection, Pfcrt and Pfmdr1 genes of isolates among school students within the age range of 11-22 years from four selected rural communities of Kwara state were studied. One hundred and eighty seven subjects (187) were selected for the study. Blood samples were collected by finger prick method for malaria screening. Nested PCR and restriction fragment length polymorphism (RFLP) were done to detect alleles of pfcrt at codon 76 and pfmdr1 at codon 86. DNA of isolates was appropriately extracted from the filter paper blots using the methanol fixation method. Logistic regression was performed on the binary observations obtained while linear regression was conducted on the fifty (50) subjects that tested positive to malaria. Results: Out of 187 subjects screened, 26.7% (50) were positive to P. falciparum. Highest malaria parasite count of 36.4% was recorded in 14-16 years age group while 20-22 years age group had the least malaria parasite count (15.4%). The result of the studied isolates indicated that out of 50 isolates analyzed for Pfcrt gene, wild type alleles accounted for 32% (16) while mutant alleles accounted for 68% (34). Alakuko Community accounted for the least number of T76 mutant alleles 10% (5) while Apado community recorded the highest number of T76 mutant gene 22% (11). For Pfmdr1 gene analysis at codon 86, isolates from Apado community showed the highest mutant type alleles (Y86) of 22% (11), while Igbonla community in Ifelodun local government had the least mutant alleles, 6% (3). Conclusion: The overall result revealed existence of mutant alleles in both the Pfcrt and Pfmdr1 genes which was higher than the wild type gene in both cases. The presence of chloroquine resistance genes among the studied population implies that alternative antimalaria drugs should be designed by pharmaceutical industry.

Antimalaria and Hematological Properties of Ethanol Leaf Extract of Pennisetum purpureum on Plasmodium berghei Infected Mice

Through bite from a female Anopheles mosquito, Malaria is transmitted by infection with single-celled parasites of the genus Plasmodium. Studies have shown it to be characterized by periodic bouts of severe chills, accompanied with high fever. It has been suggested that Pennisetum purpureum possess antiplasmodial effects, however, no scientific record(s) yet exist(s) to validate this claim. This study was therefore undertaken to determine the anti-malaria and haematological properties of ethanol leaf extract of P. purpureum in Plasmodium berghei -infected mice. Thirty-Five (35) albino mice (20g) were procured, acclimatized (for two weeks) and assigned to five groups of 7 mice each. With group I receiving standard rat feed ad-libitum (control), Groups II through V were respectively infected with Plasmodium berghei (malaria infected, untreated), Plasmodium berghei infected + treated with 5mg/kg body weight of Artesunate (malaria infected, Artesunate treated), infected with Plasmodium berghei + treated with 200mg/kg body weight of Pennisetum purpureum (malaria infected, low dose extract treated), and infected with Plasmodium berghei + treated with 400mg/kg body weight of Pennisetum purpureum (malaria infected, high dose extract treated). After 21 days of administration, mice were sacrificed, blood samples collected, centrifuged for 10 minutes at 300g, and resulting supernatant biochemically analysed for hematologic changes. Result showed a significant increase in initial parasite count across groups except control. Administration of Artesunate also caused a significant (p < .05) reduction in parasite counts upon comparison with control. More so, administration of low and high dose extract caused a significant (p < .05) reduction in parasite count following comparison with control. Administration of 200mg/kg caused the highest parasitemia suppression than high dose. We recommend for further evaluation of the plant in other to identify active ingredients responsible for the observed antimalarial activity.