Parasite Counts Research Articles

In vitro evaluation of CRISPR PX-LmGP63 vector effect on pathogenicity of Leishmania major as a primary step to control leishmaniasis

Cutaneous leishmaniasis (CL) is caused by intracellular obligate parasites (Leishmania spp.) carried by the blood-sucking of female sandflies and transmitted between mammalian hosts. Despite the high incidence and prevalence of Leishmania cases in many countries, it has been a neglected tropical disease. The current treatment approaches are limited by the complications such as loss of fertility and drug resistance. It is, therefore, essential to find new medicines to treat leishmaniasis. CRISPR/Cas9 as a powerful genome-editing tool provides the opportunity to create precise genetic manipulation to investigate the molecular basis of different leishmaniasis cases. Therefore, our main goal was to evaluate the CRISPR PX-LmGP63 vector effect on pathogenicity of Leishmania majorin vitroto challenge for using CRISPR/Cas9 as a therapeutic CL through the reduction of L. major pathogenicity by manipulating the GP63 gene. In this study, L. major parasites were transfected with CRISPR/Cas9 vectors constructed by electroporation and then added to macrophage cells on RPMI. The effect of CRISPR/Cas9 constructs on GP63 mutation, viability, and status of L. major was investigated by counting phagocytic parasites into macrophages and DNA sequence analysis. Our data validate that the use of CRISPR/Cas9 in L. major creates a new stop codon and disrupts the frame sheet of the gene by creating a new insertion (thymine), which prevents its expression. In addition, the parasite count was significantly different in the case and control of infected macrophages (P < 0.05). This study shows the successfully targeted manipulation of the L. major GP63 gene via the adaptation of the CRISPR/Cas9 editing tool. The manipulation of GP63 revealed a reduction in the infection load compared to wild-type parasite infection. Therefore, more studies are necessary for this field to help achieve a new method for the prevention and treatment of CL disease.

Plasmodium falciparum clearance time in Malawian children with cerebral malaria: a retrospective cohort study

BackgroundStandard treatment for both uncomplicated and severe malaria is artemisinin derivatives. Delayed parasite clearance times preceded the appearance of artemisinin treatment failures in Southeast Asia. Most worldwide malaria cases are in sub-Saharan Africa (SSA), where clinically significant artemisinin resistance or treatment failure has not yet been detected. The recent emergence of a resistance-conferring genetic mutation in the Plasmodium falciparum parasite in Africa warrants continued monitoring throughout the continent.MethodsAn analysis was performed on data from a retrospective cohort study of Malawian children with cerebral malaria admitted between 2010 and 2019 to a public referral hospital, ascertaining parasite clearance times across years. Data were collected from patients treated for severe malaria with quinine or artesunate, an artemisinin derivative. Parasite density was determined at admission and every subsequent 6 h until parasitaemia was below 1000 parasites/µl.The mean parasite clearance time in all children admitted in any one year was compared to the parasite clearance time in 2014, the first year of artesunate use in Malawi.ResultsThe median population parasite clearance time was slower from 2010 to 2013 (quinine-treated patients) compared to 2014, the first year of artesunate use in Malawi (30 h (95% CI: 30–30) vs 18 h (95% CI: 18–24)). After adjustment for admission parasite count, there was no statistically significant difference in the median population parasite clearance time when comparing 2014 with any subsequent year.ConclusionMalaria parasite clearance times in Malawian children with cerebral malaria remained constant between 2014 and 2019, arguing against evolving artemisinin resistance in parasites in this region.

New Insights Into Blue Light Phototherapy in Experimental Trypanosoma cruzi Infection.

The search for an effective etiologic treatment to eliminate Trypanosoma cruzi, the causative agent of Chagas disease, has continued for decades and yielded controversial results. In the 1970s, nifurtimox and benznidazole were introduced for clinical assessment, but factors such as parasite resistance, high cellular toxicity, and efficacy in acute and chronic phases of the infection have been debated even today. This study proposes an innovative strategy to support the controlling of the T. cruzi using blue light phototherapy or blue light-emitting diode (LED) intervention. In in vitro assays, axenic cultures of Y and CL strains of T. cruzi were exposed to 460 nm and 40 µW/cm2 of blue light for 5 days (6 h/day), and parasite replication was evaluated daily. For in vivo experiments, C57BL6 mice were infected with the Y strain of T. cruzi and exposed to 460 nm and 7 µW/cm2 of blue light for 9 days (12 h/day). Parasite count in the blood and cardiac tissue was determined, and plasma interleukin (IL-6), tumoral necrosis factor (TNF), chemokine ligand 2 (CCL2), and IL-10 levels and the morphometry of the cardiac tissue were evaluated. Blue light induced a 50% reduction in T. cruzi (epimastigote forms) replication in vitro after 5 days of exposure. This blue light-mediated parasite control was also observed by the T. cruzi reduction in the blood (trypomastigote forms) and in the cardiac tissue (parasite DNA and amastigote nests) of infected mice. Phototherapy reduced plasma IL-6, TNF and IL-10, but not CCL2, levels in infected animals. This non-chemical therapy reduced the volume density of the heart stroma in the cardiac connective tissue but did not ameliorate the mouse myocarditis, maintaining a predominance of pericellular and perivascular mononuclear inflammatory infiltration with an increase in polymorphonuclear cells. Together, these data highlight, for the first time, the use of blue light therapy to control circulating and tissue forms of T. cruzi. Further investigation would demonstrate the application of this promising and potential complementary strategy for the treatment of Chagas disease.

Threshold models using Gibbs sampling and machine learning genomic predictions for skin fluke disease recorded under field environment in yellowtail kingfish Seriola lalandi

Ectoparasitic infestations, such as skin fluke due to Benedenia seriolae, represent substantial fish health and welfare challenges that have caused significant industry implications. Unfortunately, existing methods used to control this disease such as bath-administered compounds or therapeutic (dietary) trials are labour intensive, costly and only temporarily effective. Genetic (genomic) methods may provide sustainable long-term solutions to improve host resistance to parasitic diseases in marine finfish species. This study thus explored possibilities for applying genomic selection to improve resistance to B. seriolae and deformity in yellowtail kingfish Seriola lalandi. Specifically, we used a total of 752 animals with individual records and 14,448 single nucleotide polymorphisms (SNPs) developed de novo from Diversity Arrays technology (DArT, a combination of genome complexity reduction methods and next generation sequencing platforms) to assess accuracy of genomic predictions for the incidence of skin fluke and deformity recorded under field (farm) condition. Our multi-locus linear and threshold mixed model analyses showed that heritabilities for the incidence of skin fluke and deformity were small (mean range h2 = 0.02–0.03). Genetic correlations of skin fluke with body weight and deformity were not significant. Our threshold Gibbs sampling and machine learning models revealed that the accuracies of genomic prediction were low for both traits (mean range from 0.151–0.432). Imputation of missing genotypes improved the prediction accuracies for skin fluke and deformity by 0.5–13.2%. Multi-trait analyses outperformed single trait models, only for deformity. Our findings suggest that genomic selection for reduced skin fluke and deformity, albeit possible in yellowtail kingfish, genetic progress made for these traits may be slow because of the low prediction accuracies across models used in this study. To enable the application of genomic selection for skin fluke or disease resistant traits recorded under field condition, it is necessary to sequence a larger number of individuals and families as well as to develop large-scale routine data recording of new phenotypes (e.g., parasite counts) to increase the prediction accuracies for these traits in the breeding program of yellowtail kingfish S. lalandi.

Red Cell Exchange as Adjunctive Therapy for Babesiosis: Is it Really Effective?

Human babesiosis is a parasitic disease prevalent in the Northeastern and Midwestern United States (US). Treatment with antibiotics is the standard of care but red cell exchange (RCE) has been used as an adjunctive treatment in more severe disease. Data for the efficacy of RCE in the treatment of babesiosis has been based on case reports and case series. An English language literature search was conducted for cases of babesiosis treated with RCE since 1980 and relevant laboratory and clinical outcome data were extracted. Similar data were obtained on severe cases of babesiosis referred for RCE in our hospitals in the time period 2000 to 2020. Fifty reports including forty-one individual case reports and nine case series were retrieved. There were 108 patients that underwent RCE with an overall mortality rate of 20%. Some patients had more than one RCE. The patients varied in the level of anemia and evidence of compromise of renal or pulmonary function. The pre-RCE level of parasitemia varied between 1.7% to 85% with the vast majority >10%. The post-RCE level of parasitemia varied between 1% to 10%. Since 2000, 32 patients were referred for RCE in our hospitals and RCE was performed on 23 of 32. There were more patients treated with RCE in the second decade as compared to the first decade, 19 versus 4 respectively. The overall mortality was 22% similar to the national data. Comparing the cohort treated with RCE to the 9 patients who were treated only with antibiotics, there were similar levels of parasitemia and laboratory parameters. The overall number of days needed to achieve a parasite count <1% was similar between the two cohorts and mortality for the antibiotics only cohort was 0%. More than 40 years after the first reported case of RCE in severe babesiosis it cannot be concluded that this adjunctive therapy favorably influences the clinical outcome. Since there is largely equipoise, a registry of severe patients treated with or without RCE could identify a benefit or otherwise.

A New Era in Malaria Diagnosis and Surveillance Using an Automated Analyzer

▪Malaria remains a global health threat with 91 countries still endemic for the disease. Despite a recent substantial decline in morbidity and mortality, the magnitude of disease burden is still enormous as indicated by the World Health Organisation (WHO) report of an estimated 212 million new cases and approximately 429 000 deaths in 2015. Sub-Saharan Africa bears the brunt of the disease with 92% of all deaths. Plasmodium falciparum is the most lethal of the 5 parasite species that infect humans, and causes 99% of all deaths. In patients infected with P. falciparum , parasitemia can dramatically escalate every 48 hours or less and can be rapidly fatal if left untreated. Accurate, timely diagnosis is thus a critical first step in the management of malaria and the WHO recommends that all suspected cases must have diagnostic confirmation either by microscopy or rapid diagnostic testing (RDT), prior to initiation of artemisinin combination therapy. Microscopic assessment of peripheral blood is the current gold standard but is subjective and requires a high level of skill, whereas RDTs indirectly detect parasite antigen and can potentially give false positive/negative results. Other methods include PCR and the detection of monocytes with ingested hemozoin, which is a surrogate marker for malaria parasites.This study describes novel technology in the form of an automated Sysmex XN-30 analyzer, which utilizes a blue laser and flow cytometry to detect and count malaria-infected red blood cells. To evaluate the performance of the XN-30, 189 residual EDTA blood samples from suspected malaria cases referred for routine diagnosis by microscopy and RDT, were analyzed within 24 hours of collection. A total of 127 P. falciparum positive samples and 62 negative samples were processed over a 6-month period. Discrepancies between the analyzer and microscopy/RDT were resolved by PCR using dried blood spots prepared at the time of sample reception. The analyzer showed excellent specificity (98.5%) and sensitivity (98.4%). Measurements were reproducible with a precision of 1.24% and showed no carryover between samples. The percentage parasitemia that was accurately detected ranged from 0.003 to 27%. Positive samples were stored at room temperature and 4°C up to 7 days and daily measurements indicated that the parasite count was stable. To exclude interference by non-malaria factors, separate malaria negative samples (20 per index) with low hemoglobin, high reticulocyte count, thrombocytopenia and hemoglobinopathies (thalassemia, sickle cell anemia) were analyzed, but these had no effect on the malaria gates. Other Plasmodium species were also detected, including P. vivax, P. ovale and P. malariae.The direct detection and quantitation of Plasmodium parasites with the XN-30 holds great promise and has many advantages over microscopy/RDT/PCR. It is automated, easy to use and provides rapid, robust and objective diagnosis of malaria, which will ensure prompt treatment and thus undoubtedly improve the quality of healthcare and alleviate the disease burden in endemic countries. Since the XN-30 provides a CBC with each analysis, it may also serve as a valuable diagnostic aid in detecting unsuspected cases with fever of unknown origin. This scenario is becoming more common as international travel to endemic countries increases and tourists often present with symptoms when returning to their home country, where clinicians may not have a high index of suspicion for malaria, but would request a CBC as part of a diagnostic workup. The decline in parasitemia can be monitored by the XN-30 and if the clinical response to therapy is delayed, this may serve as an early warning sign of drug resistance. This will yield valuable data on the spread of artemisinin resistance, which is alarmingly prevalent in the Greater Mekong sub-region. The analyzer can differentiate gametocytes making it useful for detection of asymptomatic carriers who can be treated to prevent the transmission of parasites to the mosquito vector and thus protect communities. This is a vital step in disrupting the lifecycle of the parasite and is in line with global initiatives to eliminate malaria. It can also process finger prick samples making it suitable for infants and population studies, as well as monitoring efficacy of vaccines or new drugs in clinical trials. The XN-30 is currently for research use only but further development and clinical studies are in progress. DisclosuresPillay:Sysmex Corporation: Research Funding. Coetzer:Sysmex Corporation: Research Funding.

On the trail of detecting genetic (co)variation between resistance to parasite infections (Diplectanum aequans and Lernanthropus kroyeri) and growth in European seabass (Dicentrarchus labrax)

The European seabass is one of the main commercial fish produced in Mediterranean marine aquaculture. Recently, its production has been negatively affected by losses due to frequent and recurring outbreaks of parasitic, bacterial and viral diseases. In recent years, the gill parasites Diplectanum aequans and Lernanthropus kroyeri are increasingly becoming more dominant and contributing more significantly to the observed losses. Genetic improvement for disease resistance represents an important strategy for controlling infectious diseases in farmed fish by increasing their robustness. In order to determine the possibility of including such trait in selective breeding programs, we need to comprehend whether additive genetic variation for resistance against Diplectanum aequans and Lernanthropus kroyeri exists. For this purpose, two open-sea parasite cohabitation trials (for two consecutive years) were performed in the commercial production sites of a private company (Nireus S.A), that had high infestation with Diplectanum aequans and Lernanthropus kroyeri. Juvenile European seabass (9425 offspring from 91 full-sib and half-sib families per year), originating from the company's breeding program were equally divided into two groups and transferred to two commercial farming sites located in the areas of Nafpactos and Sagiada, in western Greece, for the intended cohabitation studies. The parasite numbers on all the gill arches were counted and recorded at the end of the trials for the infestation levels. A third site (Palairos) without any parasite infestation was used as a control site. A multi-trait animal model was used to estimate the variance-covariance components and to evaluate the genetic parameters for Parasite Counts, recorded from all the gill arches of the fish, and their corresponding growth in sea cages. The estimated heritabilities for parasite count, using untransformed data, were 0.20 (D. aequans) and 0.28 (L. kroyeri) and for transformed data 0.29 and 0.26, respectively. The heritability estimates for body weight were 0.42−0.51 for D. aequans and 0.28−0.51 for L. kroyeri trials. Similarly, estimated heritabilities for the growth in sea cages were 0.43 and 0.29, respectively. Although parasite count has a low to medium unfavorable genetic correlation with body weight and growth (0.09−0.37), it seems that it is not significantly impairing selection for growth. Furthermore, the results of this study are very promising in terms of the existing potential for genetic improvement of parasite resistance, and provides a good basis for further genetic analysis using molecular markers.

Parasite counts or parasite incidences? Testing differences with four analyses of infracommunity modelling for seven parasite-host associations.

One of the challenges in studies of parasite community ecology is whether the input data for analyses should be parasite abundances/counts, i.e. count data (CD), or parasite incidences (presences/absences), i.e. incidence data (ID). We analysed species responses to environmental factors and species associations in the infracommunities of helminths and ectoparasites in four hosts from Europe (Sorex araneus and Myodes glareolus) and South Africa (Rhabdomys pumilio and Rhabdomys dilectus) and compared the results of four analyses [redundancy analysis (RD), RLQ analysis, joint species distribution modelling (JSDM) and Markov random fields (MRF)] that used either CD or ID as an input. In addition, we compared the differences between the CD and ID results of two analyses (JSDM and MRF) across parasite species between (a) host species within helminths and ectoparasites; (b) helminths and ectoparasites within a host species; and (c) parasite species with contrasting levels of intensity. The results of most analyses for the majority of parasite-host associations were qualitatively similar. However, models based on the ID input performed better than models based on the CD input in three out of four types of analyses (RDA, JSDM and MRF). The differences between the CD and ID models varied between host species (being the lowest in R. pumilio for JSDM and in S. araneus for MRF). However, they were not affected by the level of parasite intensity.

Comparison of capillary, venous and buffy coat blood samples in detecting Plasmodium species among malaria suspected patients attending at Hamusite health center. A cross-sectional study

BackgroundBoth capillary and venous blood samples have been interchangeably used for the diagnosis of malaria in Ethiopia. However, Plasmodium parasites are thought to be more concentrated in capillary than in venous blood. Hence, selecting a sample source where parasites are more concentrated is indispensable approach in order to maximize the accuracy of blood film microscopy. Therefore, the present study aimed to compare the detection rate and the parasitemia level of Plasmodium species from conventional capillary and venous blood films, and buffy coat preparations.MethodsA facility based cross-sectional study was conducted from Feburary to March 2020 among 210 febrile patients attending Hamusite health center, northwest Ethiopia. Capillary and venous blood samples were collected and buffy coat was prepared from each sample. Thin and thick blood films were prepared, stained, and examined microscopically following standard protocol. Data were analysed using Statistical Package for Social Sciences Software version 20 and Med-Calc software version 19.3.ResultsCapillary blood buffy coat (61/210, 29.0%) had significantly higher detection rate as compared to capillary (48/210, 22.9%) and venous (42/210, 20.0%) blood films (p < 0.001). However, no significant difference was observed between capillary and venous blood films (p = 0.070) in detecting Plasmodium species. The highest and the lowest mean asexual stage parasite counts were found in capillary blood buffy coat (4692.88) and venous blood (631.43) films, respectively showing significant variations (p < 0.001). Mean gametocyte count was also highest in capillary blood buffy coat (3958.44). As compared to capillary blood buffy coat, the sensitivity of venous blood buffy coat, capillary blood film and venous blood film were 73.8, 78.7, 68.9%, respectively.ConclusionCapillary blood buffy coat samples showed the highest sensitivity in detecting and quantitating malaria parasites that its use should be promoted in clinical settings. However, conventional capillary and venous blood films could be used interchangeably.

In vitro and in vivo trypanocidal activity of a benzofuroxan derivative against Trypanosoma cruzi

Chagas disease, caused by Trypanosoma cruzi, is a major public health problem and is described as one of the most neglected diseases worldwide. It affects about 6–7 million people. Currently, only two drugs are available for the treatment of this disease: nifurtimox and benznidazole. However, both drugs are highly toxic and have several side effects, which lead many patients to discontinue treatment. Moreover, these compounds show a significant curative efficacy only in the acute phase of the disease. Therefore, searching for new drugs is necessary. The objective of this study was to evaluate the in vitro and in vivo activity of a benzofuroxan derivative (EA2) against T. cruzi, and to evaluate the hematological and biochemical changes induced by its treatment in animals infected with T. cruzi. The results were then compared with those of healthy controls. In vitro testing was first performed with T. cruzi epimastigote forms. In this experiment, EA2 was diluted at three different concentrations (0.25, 0.50, and 1%). In vitro evaluation of the trypanocidal activity was performed 24, 48, and 72 h after incubation. In vivo assays were performed using three different doses (10, 5, and 2,5 mg/kg). Mice were divided into 10 groups (five animals/group), wherein four groups comprised non-infected animals (A, G, H, I) and six groups comprised infected animals (B, C, D E, F, J). Groups B and J represented the negative and positive controls, respectively. Groups G, H, and I were used to confirm that EA2 was not toxic to non-infected animals. Parasitemia was measured in infected animals and the hematological and biochemical profiles (urea, creatinine, albumin, aspartate aminotransferase, alanine aminotransferase, and alkaline phosphatase) were evaluated in all animals. EA2 demonstrated in vitro trypanocidal activity at all concentrations tested. Although it did not demonstrate a curative effect in vivo, EA2 was able to retard the onset of parasitemia, and significantly reduced the parasite count in groups D and E (treated with 5 and 2.5 mg/kg, respectively). EA2 did not induce changes in hematological and biochemical parameters in non-infected animals, demonstrating that it is not toxic. However, further assessments should aim to confirm the safety of EA2 since this was the first in vitro and in vivo study conducted with this molecule.

Retraction Note: Antimalarial efficacy of Pongamia pinnata (L) Pierre against Plasmodium falciparum (3D7 strain) and Plasmodium berghei (ANKA)

The objective of the current study was to assess the in vitro antiplasmodial activities of leaf, bark, flower, and the root of Pongamia pinnata against chloroquine-sensitive Plasmodium falciparum (3D7 strain), cytotoxicity against Brine shrimp larvae and THP-1 cell line. For in vivo study, the plant extract which has shown potent in vitro antimalarial activity was tested against Plasmodium berghei (ANKA strain). The plant Pongamia pinnata was collected from the herbal garden of Acharya Nagarjuna University of Guntur district, Andhra Pradesh, India. Sequentially crude extracts of methanol (polar), chloroform (non-polar), hexane (non-polar), ethyl acetate (non-polar) and aqueous (polar) of dried leaves, bark, flowers and roots of Pongamia pinnata were prepared using Soxhlet apparatus. The extracts were screened for in vitro antimalarial activity against P. falciparum 3D7 strain. The cytotoxicity studies of crude extracts were conducted against Brine shrimp larvae and THP-1 cell line. Phytochemical analysis of the plant extracts was carried out by following the standard methods. The chemical injury to erythrocytes due to the plant extracts was checked. The in vivo study was conducted on P. berghei (ANKA) infected BALB/c albino mice by following 4-Day Suppressive, Repository, and Curative tests. Out of all the tested extracts, the methanol extract of the bark of Pongamia pinnata had shown an IC50 value of 11.67 μg/mL with potent in vitro antimalarial activity and cytotoxicity evaluation revealed that this extract was not toxic against Brine shrimp and THP-1 cells. The injury to erythrocytes analysis had not shown any morphological alterations and damage to the erythrocytes after 48 h of incubation. Because methanolic bark extract of Pongamia pinnata has shown good antimalarial activity in vitro, it was also tested in vivo. So the extract had exhibited an excellent activity against P. berghei malaria parasite while decrement of parasite counts was moderately low and dose-dependent (P < 0.05) when compared to the control groups, which shown a daily increase of parasitemia, unlike the CQ-treated groups. The highest concentration of the extract (1000 mg/kg b.wt./day) had shown 83.90, 87.47 and 94.67% of chemo-suppression during Suppressive, Repository, and Curative tests respectively which is almost nearer to the standard drug Chloroquine (5 mg/kg b.wt./day). Thus, the study has revealed that the methanolic bark extract had shown promisingly high ((P < 0.05) and dose-dependent chemo-suppression. The phytochemical screening of the crude extracts had shown the presence of alkaloids, flavonoids, triterpenes, tannins, carbohydrates, phenols, coumarins, saponins, phlobatannins and steroids. The present study is useful to develop new antimalarial drugs in the scenario of the growing resistance to the existing antimalarials. Thus, additional research is needed to characterize the bioactive molecules of the extracts of Pongamia pinnata that are responsible for inhibition of malaria parasite.

Oral acute toxicity and antimalarial potentials of aqueous and methanolic extracts of roots, leaves and stem of Dictyandra arborescens (Welw.) on Plasmodium berghei infected mice

BackgroundMalaria is one of the tropical diseases of universal concern particularly with continuous appearance of resistant strains of P.falciparum. This calls for continous screening of traditional plants such that new and effective antimalarial agents will be developed. This study therefore explored the oral acute toxicity and antimalarial potentials of aqueous and methanolic extracts of roots, leaves and stem of Dictyandra arborescens on Plasmodium berghei infected mice.ResultsNo mortality was recorded in any of the experimental animal groups even at the highest tested dose (5000 mg/kg b.wt) of the extract after monitoring them for 4hrs and subsequently for 7 days. Out of the six extracts, methanolic extracts of the roots and leaves exhibited more antimalarial activity than others. A significant difference (P < 0.05) was statistically observed in the parasite count of groups that received methanol extracts of roots and leaves of D. arborescens. This observation was made when these two extracts were compared with other groups as well as the negative control. However, activity of the standard antimalarial drug (artesunate) was higher (p˂0.05) than those of the extracts. Phytochemicals such as tannins, alkaloids, saponins, terpenoids, flavonoids etc. were present in the extracts in varying quantities. GC–MS analysis of methanol extract of the root of this plant showed different chemical compounds.ConclusionAdministration of aqueous and methanol extracts of roots, leaves and stem of D. arborescens in mice is not harmful at any dose less than or equal to 5000 mg/kg. Methanol extracts exhibited more antimalarial activity than aqueous extracts suggesting that antimalarial activity of the plant parts could be affected by the solvent used for extraction and antimalarial activity may be more in a particular part of a plant. The presence of different bioactive compounds identified in phytochemical and GC–MS analysis could be the fundamental scientific evidence for the antimalarial activity exhibited by this plant especially in the root.

Kangaroos at maximum capacity: health assessment of free-ranging eastern grey kangaroos on a coastal headland

Sprawling urban development is fragmenting the landscape and native wildlife habitats on the Australian east coast. The impact of this rapid urbanization on wildlife health is largely unknown. This study surveyed the health of a high-density (5.4 individuals per ha) population of eastern grey kangaroos (Macropus giganteus) affected by urban encroachment and prolonged drought. Blood parameters (hematological and serum protein), trace element and heavy metal concentrations, and parasite counts (fecal worm egg counts, ticks, and mites) are reported for a sample of ≤ 54 kangaroos at Look at Me Now Headland, New South Wales, Australia. These parameters were compared to lower density kangaroo populations from other sites in New South Wales. We found the health and welfare of this population to be severely compromised, with nonregenerative anemia and nutritional deficiencies evident. Our results indicate that high-density kangaroo populations isolated by urban encroachment are at significant health risk. To prevent further decline in this population’s health, we discuss management strategies that could be employed, concurrent with ongoing health and disease monitoring, to mitigate the poor health outcomes in this population. We conclude that it is essential to retain habitat connectivity when altering land use in areas with resident kangaroo populations if managers are to maintain healthy populations.

Prevalence and Faecal Egg Counts of Gastrointestinal Parasites of Merino Sheep in Lesotho

The present study aimed to evaluate the effect of the agroecological zone, host age, and gender on the prevalence and faecal egg load of gastrointestinal parasites (GIPs) for six months (July to December) in the Maseru and Quthing districts, Lesotho. A total of 1919 faecal samples were examined using the McMaster technique. The data were analyzed through generalized estimating equations (GEE) under the binary logistic regression model to determine the significant differences for the GIPs prevalence. Moreover, faecal egg counts (FEC) data were analyzed for repeated measures using GEE. In total, three types of GIPs, namely nematodes, coccidia, and cestodes were identified in this study. The overall prevalence rates of nematodes, coccidia, and cestodes were 53.9%, 46.5%, and 4.3% in the Maseru district, respectively. Furthermore, the Quthing district indicated the prevalence rates of 65.0%, 38.2%, and 0.9% for nematodes, coccidia, and cestodes, respectively. In the Maseru district, the overall faecal egg counts for nematodes, coccidia, and cestodes were within the ranges of 0-20.3, 0-90, and 0-600 eggs per gram, respectively. Additionally, the faecal egg counts in the Quthing district ranged from 0 to 8.000, 6.700, and 2.000 eggs per gram for nematodes, coccidia, and cestodes, respectively. The majority of the Merino sheep (&gt;69%) in both districts had lower faecal egg counts (100-800) per gram. The agroecological zone affected the nematode infestation in both districts. Coccidia in the Quthing was higher in the mountain areas. In the Maseru district, the nematode infestation was not age-dependent; however, in the Quthing district, the prevalence was higher in juveniles, compared to adults. Age and gender did not affect the prevalence and faecal egg counts of nematodes and coccidia. The coccidian faecal egg loads were higher in females, compared to males. Merino sheep in Lesotho are mostly infected with gastrointestinal nematodes and protozoal coccidia, which could have a tremendous impact on their health and productivity. It is, therefore, of significant importance to develop the deworming strategy for sheep of different age and gender groups in different agroecological zones.

PREVALENCE OF ZOONOTIC CRYPTOSPORIDIUM SPP. ISOLATES IN NJORO SUB-COUNTY, NAKURU COUNTY, KENYA.

Background:There is no information on human and animal Cryptosporidium spp. in Njoro sub- county. The risk posed to humans and animals within the sub-county is therefore unknown.Materials and Methods:A total of 1476 animal and 378 human fecal samples were evaluated. Multivariate logistic regression was used to evaluate association between infection status and the predisposing factors. Results were expressed as odds ratio (OR) with a 95% confidence interval. Chi-square and Maentel–Haenszel tests were used to quantify relationships among variables.Results:Prevalence of Cryptosporidium spp. was 9.8% in humans, 10.8% in cows, 19.6% in sheep and 4.5% in goats. Prevalence in humans was significantly higher in females 12/37. Infection was highest in the elderly (27.27%), and significantly lower in adolescents and adults at 8.66% and 9.59%, respectively. Goats had lowest overall parasitization at all levels, while sheep had the highest parasitization at levels (+1 and +2). Relatively, humans had the highest parasite counts +3 cases (1.5%).Conclusion:Cryptosporidium spp. is prevalent in Njoro sub-county and domestic animals are important reservoirs and a potential source of zoonosis in humans. Children, elderly and females are at increased risk of infection, especially during rainy season. The study recommends maintenance of proper sanitation when handling domestic animals, treatment of drinking water and use of alternative safer sources of water in order to reduce infection.

Combination of immunostimulants with moxidectin in the treatment of animals experimentally infected with Haemonchus contortus

Considering the importance of Haemonchus contortus infection in herds along with parasitic resistance, the goal of this study was to evaluate the influence of the administration of adjuvants alone or in combination with anthelmintics for the treatment of H. contortus, in experimentally infected sheep. Thirty sheep of the Texel breed of both genders, raised in a herd located in the subtropical region of Brazil, were used in this experiment. Experimental infection with H. contortus was performed in sheep, and the infected sheep were then separated into groups for the administration of antiparasitic and immunostimulant drugs. The results obtained from the excretion of eggs per gram of feces and the count of parasites during necropsy affirm that the use of adjuvants in combination with anthelmintics is associated with higher efficacy of treatment, lower rate of reinfection, and retardation in the development of anthelmintic drug resistance by H. contortus. Based on these results, we can conclude that the combination of anthelmintics and immunostimulants may favor potential anthelmintic treatments for H. contortus.

Drug Repurposing Patent Applications October-December 2020.

Drug Repurposing Patent Applications October-December 2020.

A tablet derived from Andrographis paniculata complements dihydroartemisinin-piperaquine treatment of malaria in pregnant mice.

The use of standard antimalarial drugs, such asdihydroartemisinin-piperaquine (DHP) for the treatment of malaria during pregnancy is limited due to the risk of teratogenicity. The alternative is therefore required although fewexist. Here we show a phytopharmaceutical drug derived from Andrographis paniculata (AS201-01), which is effective as herbal antimalarial both in vitro and in vivo and may be a suitable alternative when used in complementary treatment with DHP. Plasmodium berghei infected pregnant BALB/c mice were divided into four groups: G1 (negative control), G2 (AS201-01), G3 (DHP), and G4 (combination of DHP and AS201-01). Pheripheral blood was collected during therapy for counting parasitemia. Placental samples were analyzed for the expression of IFN-γ, TNF- α, IL-10, placental parasite counts and foetal morphology. Groups G4 and G3 both showed a 100% inhibition of peripheral parasitemia. However, the treatment in G4 was found to be less effective than that in G2 and G3 in preventing placental parasitemia. The G4 treatment was able to reduce the expression of IFN-γ and IL-10, whereas TNF-α was not significantly different from the control group. Foetal morphologic abnormalities were observed in all groups except G2; G4 showed lower percentage of abnormalities compared to G3 and G1. A combination of A.paniculata tablet (AS201-01) with DHP has the potential to reduce the toxicity of DHP in malaria treatment.

Biomarkers of disease severity in vivax malaria.

Severe complications have been observed and established for Plasmodium falciparum as well as P. vivax infections worldwide. Although P. vivax infection is not fully acknowledged as malignant malaria, recently life-threatening complications have been reported to occur in many studies. The recognition of biomarkers with excellent sensitivity and reliability plays a prime role in disease management. Acute inflammatory response and oxidative stress are observed in malaria due to the production of reactive oxygen species. Lipid and protein oxidative injuries are prospective biomarkers for disease severity owing to the damage caused by the parasite. We have tried to find out whether protein carbonylation (PC), lipid peroxidation (LPO) and superoxide dismutase (SOD) could suffice as a biomarker for severe vivax malaria or not. Blood samples were collected from the individuals attending Jawaharlal Nehru Medical College of Aligarh Muslim University during the wet season of malaria transmission. Microscopy and rapid diagnostic kits were used as a tool for malaria diagnosis. A total of 214 subjects were enrolled for the study: 30 febrile controls and 184 subjects with vivax malaria. Protein carbonylation and lipid peroxidation were found to be directly associated with parasite count and total antioxidant status (TAS). Increase in oxidative stress was also observed in severe vivax malaria patients. Levels of uric acid and bilirubin too were raised in complicated cases. Protein carbonylation was found to be a more reliable indicator of vivax malaria severity than lipid peroxidation.

Prevalence and fecal egg load of gastrointestinal parasites of Angora goats in four agro-ecological zones in Lesotho.

Background and Aim:Goats are reared for their meat, mohair and other socio-cultural needs in Lesotho. Helminth infections are some of the major setbacks in the goat production industry due to their negative impact on animals’ health, resulting in significant losses on meat and mohair production and death. A cross-sectional study was conducted to determine the prevalence, fecal egg infestation, and morphological identification of gastrointestinal parasites in goats.Materials and Methods:Fecal samples were collected from 765 goats and subjected to McMaster egg counting techniques using the flotation method. Statistical analyses were performed using the Statistical Package for the Social Sciences (SPSS v.26.0).Results:The overall prevalence of gastrointestinal parasites was 94.7%, and the identified gastrointestinal parasites were nematodes (64.7%), coccidia (25.8%), and cestodes (4.2%). Haemonchus contortus was identified as the prevalent gastrointestinal nematode species found in goats. The prevalence and fecal egg count of gastrointestinal parasites were significantly higher (p<0.05) in goats located in the highlands and Senqu River Valley, while goats in the lowlands demonstrated a significantly (p<0.05) higher prevalence of H. contortus. Immature goats and kids were more significantly (p<0.05) prone to gastrointestinal parasites.Conclusion:The nematodes and coccidia infestations were prevalent in goats located in the highlands and foothills, respectively, whereas nematode and coccidia fecal egg loads were higher in goats located in the foothills and Senqu River Valley, respectively.