Giemsa-stained Blood Smears Research Articles

Hematological alterations in large Babesia species infection in dogs of Kannur District of Kerala.

Canine babesiosis is a tick-borne disease caused by apicomplexan intraerythrocytic hemoprotozoan parasites. It is caused by the small (Babesia gibsoni, B. conradae, and B. vulpes) and large (B. vogeli, B. canis, and B. rossi) Babesia groups. As per the recent reports, the most prominent Babesia species encountered in the Kerala state are the small Babesia, B. gibsoni followed by the large Babesia, B. vogeli. The latter is regarded as mildly pathogenic, causing subclinical or mild disease; however severe complications like systemic inflammatory response syndrome, multiple organ dysfunction syndrome, etc. have also been reported. The information on the status of hematological alterations in naturally infected dogs with large Babesia is lacking, particularly from the state of Kerala. The present study involves a retrospective study of clinical cases of large Babesia infection in dogs. The complete haematological profile from well-documented laboratory records of 4039 dogs suspected for babesiosis presented to District Veterinary Centre, Kannur during the period from December 2018 to October 2020 was analyzed for the study. Natural infections were recorded in 35 (0.87%) dogs based on the presence of intraerythrocytic piroplasm of large Babesia spp. by light microscopic examination of Giemsa-stained blood smears. The most consistent features observed were mild to moderate regenerative, normocytic and normochromic anemia, lowered to normal neutrophil count and thrombocytopenia. In comparison to hemolytic anemia, thrombocytopenia was the most frequent clinicopathological finding in the study with an increased presence of large activated platelets or macro-platelets.

Mixed Infection of Tick-borne Haemo-parasites in Water Buffalo and Associated Pathological Responses and Treatment

Background: Water buffaloes are important milch species of tropical and subtropical countries. In general, they are considered to more resistant to diseases compared to crossbred cattle. However, a recent problem of tick-borne diseases in the water buffaloes by the field veterinarians and farmers causing production losses was observed. The study was conducted to explore the spectrum of tick-borne diseases (TBDs) infections in buffaloes and analyze the associated risk factors. Methods: Acute-phase response, cytokine and oxidative stress in infected buffaloes with TBDs were evaluated and compared with the negative buffaloes (control) to elucidate their role in pathogenesis and outcome of infection. The study was undertaken in 107 tick-infested water buffaloes. The conventional Giemsa stained blood smear (GSBS) based confirmation and classification of infection of haemo-parasites were made. The statistical model was used to understand their relevance with TBDs. Conclusion: Tick-borne disease in water buffaloes must be looked upon seriously to maintain good productivity. The buffaloes are often accompanied by low and un-yielding clinical symptoms due to associated co-infections of haemo-parasites. Our study showed that the buffaloes had a high prevalence (47.66%) of haemo-parasites transmitted by the ticks associated with co-infections and a low level of parasitemia. Most buffaloes responded symptomatically with the different modalities administered. Production parameter was not restored post 15 days of treatment. Oxidative stress is one of the important mechanisms of production losses in infected buffaloes.

Hematologic values in domestic pigeons naturally infected with Mycobacterium avium subsp. avium.

Avian mycobacteriosis is an important disease of birds and is most often caused by Mycobacterium avium or Mycobacterium genavense. However, little information on the hematologic changes associated with this infectious disease in pigeons has been published. The aim of this investigation was to compare the hematologic parameters of domestic pigeons (Columba livia var. domestica) naturally infected with Mavium subsp. avium (MAA) with clinically healthy pigeons. Blood samples were collected from 12 pigeons with suspected mycobacteriosis and 12 clinically healthy pigeons. All the birds with suspect infections were necropsied, and affected organs were cultured and examined on histopathology for mycobacteriosis. Total leukocyte and erythrocyte counts were performed on each blood sample with the Natt and Herrick method using a Neubauer hemocytometer. White blood cell (WBC) differential counts were performed on Giemsa-stained blood smears. Packed cell volumes (PCVs) were measured using the microhematocrit technique. Hemoglobin concentrations were measured with a spectrophotometer using the cyanomethemoglubin method. Mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentrations (MCHCs), and mean cell volumes (MCVs) were calculated manually. All of the infected birds had typical histopathologic findings of avian mycobacteriosis, which were confirmed using microbiologic and molecular methods to detect MAA. The hematologic data from the two groups were compared. The total WBC, heterophil, lymphocyte, and monocyte counts were significantly higher, and the PCV, HGB, MCH, and MCHC values were significantly lower in the infected birds compared with the clinically healthy pigeons.

Development of a novel multiplex PCR assay for the detection and differentiation of Plasmodium caprae from Theileria luwenshuni and Babesia spp. in goats

Intraerythrocytic parasites are traditionally identified by the microscopic examination of Giemsa-stained blood smears. However, this method does not always allow for the identification of individual species in goat's RBCs. Moreover, its unreliability in detecting low levels of parasitemia makes it unsuitable for epidemiological investigations and leaves goat farms vulnerable to potential outbreaks. In the present study, a novel multiplex PCR (mPCR) targeting the cytochrome c oxidase subunit I (COI) gene was developed to detect and subsequently differentiate Plasmodium caprae, Theileria luwenshuni, and Babesia spp. The specificity of each primer set was assessed both in silico and with a panel of DNA samples from the hosts themselves and other goat hemoparasites. Amplicons generated from each pair of primers were 664, 555, and 320-bp for P. caprae, Babesia spp., and T. luwenshuni, respectively. These products were further confirmed by sequencing. Our novel mPCR reactions successfully demonstrated the accurate and simultaneous amplification of the three parasites’ DNA samples. The current mPCR method showed no cross-amplification with unintended targets. The detection limit of the mPCR in this study was 108 parasites’ DNA copies per reaction. The current mPCR was able to detect the minimum parasitemia of approximately 0.001%, 0.000005%, 0.00001% for P. caprae, Babesia spp. and T. luwenshuni, respectively. The diagnostic specificity in the detection of P. caprae and T. luwenshuni ranged from 94.9 to 100 %. The mPCR was further applied to a collection of field blood samples from five provinces in Thailand to validate its reliability and applicability. The results demonstrated the successful detection of P. caprae, Babesia spp. and T. luwenshuni in goat samples with the same sensitivity levels as conventional PCR methods. This study also confirmed the presence of T. luwenshuni and Babesia spp. in Thai goats. The current mPCR method offers an alternative for the diagnosis of P. caprae, T. luwenshuni, and Babesia spp., either single or under co-infection conditions, and for large-scale surveillance.

Prevalence of Malaria among Children under Five Years in the Niger Delta Region of Nigeria

Background: Malaria still remains an overwhelming cause of morbidity and mortality among children under five years of age, especially in sub-Saharan Africa. The aim of this study was to determine the prevalence of Plasmodium parasitemia among children below 5 years of age in Akwa-Ibom, Delta and Rivers States, located in the Niger Delta region of Nigeria.
 Methodology: A cross sectional study of 2630 children, selected by multistage sampling from Akwa-Ibom, Delta and Rivers States of Nigeria between April and June 2019. Parasitological diagnosis was by Plasmodium falciparum histidine-rich protein 2-based malaria rapid diagnostic test (RDT) and microscopy of giemsa-stained blood smears. Demographic information was collected using soft copies of pretested interviewer-administered questionnaires via the Open Data Kit application installed on android phones. Data analysis was performed using the statistical software SPSS version 25. Frequency, percentages, and Chi-square test were used to interpret data at a confidence interval of 95% and a p-value less than 0.05 was regarded as statistically significant.
 Results: A total of 2630 children less than 5 years old were included in the study, 1016 from Akwa-Ibom and 807 each from Delta and Rivers states respectively. Malaria parasitaemia was detected in 230 (8.6%) and 198 (7.4%) children by RDT and microscopy, respectively. RDT used in this survey has high diagnostic accuracy (98.8%) compared to microscopy.
 Conclusion: The study shows a decline in the prevalence of malaria in children under 5 years. It also demonstrates the reliability of the RDTs in the diagnosis of malaria. The use of RDTs is thus further recommended especially in peripheral centers where the access to skilled microscopists and laboratory infrastructure may be lacking.

Comparative epidemiology and pathophysiology of patent and latent babesiosis caused by Babesia bigemina in buffaloes and cattle from different agroclimatic zones of Punjab State, India.

To conduct comparative epidemiology of parasitologically positive (patent) and polymerase chain reaction positive (latent) cases of bovine babesiosis in Bet Region (low-lying areas adjoining Sutlej, Beas, Ravi, and Ghaggar rivers of Punjab) of diverse agroclimatic zones of Punjab state in relation to haematobiochemical parameters as patho-physiological markers, blood samples from 783 dairy animals (487 buffaloes and 296 cattle) were analysed parasitologically by Giemsa-stained blood smears (GSBS) and by molecular-based polymerase chain reaction (PCR) targeting SpeI-AvaI restriction fragment of Babesia bigemina. We ruled out the endemicity of the disease with 2.17% patent and 3.96% latent cases of B. bigemina with significantly higher prevalence (P < 0.01) in cattle than buffaloes. The spatial distribution for a guideline to local veterinary practitioners and policy-makers indicated highest number of patent and latent cases in western zone and undulating plain zone, respectively. District wise highest prevalence of patent as well as latent infection observed in SBS Nagar of undulating plain zone showed substantial agreement (Kappa value: 0.70) between the two techniques. Haematology revealed marked microcytic hyperchromic anaemia in patent animals of group I (GSBS positive; n = 17) and latent animals of group II (PCR positive; n = 14) as compared to disease-free controls (group III; n = 10). Blood urea nitrogen (BUN), creatinine and gamma-glutamyltransferase (GGT) levels were significantly increased (P < 0.05) in group I in comparison to group II and group III indicated comparative pathogenic effect of babesiosis in patent cases. Though patent cases showed higher pathogenicity of babesiosis, diagnosis of latent infection is significant as it may act as source of infection for spread to other highly prone bovines.

Influence of Gender on Some Red Cell Indices, L-Arginine and D-dimer in Malaria Parasite Severity amongst Children Resident in Rivers State, Nigeria

Aim: The aim of the study was to assess the influence of gender on malaria parasite severity in children resident in Rivers State, Nigeria.&#x0D; Study Design: The study was cross-sectional observational study.&#x0D; Place and Duration of Study: University of Port Harcourt Teaching Hospital, Rivers State, Nigeria, between the month of March and August 2020.&#x0D; Methodology: A total of 822 pediatrics (0-16 years), were randomly selected for this study after due parental consent. 5ml of venous blood was collected from each subject: 1ml was dispensed into paediatric EDTA (for haematologic and parasite density) and 4 ml into sodium citrate bottle for L-arginine assay by ELISA-method, while Full blood count was determined using haematological auto-analyser, Mindray BC-6800. Malaria density was determined by microscopic method using thick and thin Giemsa stained blood smears. Level of significance was set at P&lt;0.05.&#x0D; Results: There was a significant decrease (p&lt;.05) in the mean (41.04±3.80%) neutrophil count in female subject with complicated malaria compared with the control (42.81±0.98%) as against a significant decrease in mean (37.71±0.96%) neutrophil count of female subject with uncomplicated malaria. A significant decrease in neutrophil (28.05±3.37%) of male subjects with complicated malaria and uncomplicated malaria (36.10±0.79%) was seen when compared to control (44.32±0.88%). Again, a significant decrease (p&lt;.05) in eosinophil count of female with complicated malaria (3.32±0.74%) was seen when compared with the control subject (3.81±0.19%) and no significant difference was seen in female subjects with uncomplicated malaria (3.62±0.19%) when compared with the control. A significant increase in eosinophil was seen in male subjects with complicated malaria (4.47±0.66%) and uncomplicated malaria (4.52±0.16%) when compared with the control (3.88±0.17%). There was observed a significant difference (p&lt;.001) in the mean L-arginine values of female subjects with complicated (39.22±9.57pg/ml) and uncomplicated (65.13±2.41 pg/ml) malaria compared with the control (42.85±2.48 pg/ml). However, no significant difference was seen in male subjects with complicated (33.21±8.49) and uncomplicated (45.51±2.00 pg/ml) malaria when compared with control (47.97±2.21 pg/ml). Also, a significant difference (p&lt;0.0019) was seen between the mean D2D values of female subjects with complicated (6436.64±568.94 pg/ml) and uncomplicated (2824.55±143.46 pg/ml) malaria among the study subjects as against the control (1866.39±147.35 pg/ml).&#x0D; Conclusion: In conclusion, this study showed a trend between gender and malaria type did not significantly change haematological parameters with the exception of the immune cells such as NEU, LYM, and EOS. However, a significant increase in L-arginine among female subjects was seen indicating a faster rate of malaria clearance.

Conventional and Molecular Diagnosis of Theileriosis (Theileria annulata) in Cattle in Sulaimani Province, Northern Iraq.

Theileria annulata has been identified as a major cause of bovine theileriosis in Iraq. The research was designed to reveal the prevalence of Theileriosis (Theileria annulata) in cattle in Sulaimani province, Kurdistan Region- Iraq from the 10th of March 2021 to the 10th of April 2021. Eighty blood samples were collected. Different techniques of parasitic detection, including Microscopic and PCR amplification were used. The results of microscopic examination and PCR assay revealed that 27.5% and 31.25% cattle were infected, respectively. The microscopic examination of the Giemsa-stained blood smears of infected erythrocytes revealed the typical morphological structure of piroplasm. According to the phylogenetic analysis of the partial 18S ribosomal RNA gene, Theileria field isolates were shown to be related to other T. annulata strains, including those from Turkey, Italy, and Pakistan.This study concludes that T. annulata is the causative agent of theileriosis of cattle in Sulaimani province and suggests that PCR technique is excellent tools for epidemiological researches and control programs.

First report of Babesia occultans in two symptomatic cows in Iran.

This study reports the identification and first molecular characterization of Babesia occultans from naturally infected cows in Iran. Microscopic examination showed pyriform trophozoites, and ring-shaped merozoites (>2.5 μm) in Giemsa-stained blood smears obtained from two symptomatic cows in West-Azarbaijan province, Iran. PCR amplification of the partial 18S rRNA gene including the V4 hypervariable region were carried out on DNA extracted from blood samples. BLAST analyses of the partial 18S rRNA (approximately 400 bp) obtained from two cows revealed the presence of B. occultans and the detected sequences were identical to each other. Comparisons of the partial 18S rRNA sequence of the current isolate with other B. occultans sequences from Tunisia, South Africa, Turkey, Pakistan, and China confirmed the relation of the Iranian isolate to the species B. occultans. Sequence analysis of the obtained B. occultans showed 99.5-100% identity to the previously reported isolates. The sequences of B. occultans had 100% identity to a sequence obtained from ticks in Tunisia. This report is the beginning of a path to further research about B. occultans in vectors and reservoirs throughout Iran.

Cross-sectional survey of cattle haemopathogens in Constantine, Northeast Algeria.

This aim of the present study was to estimate the prevalence of haemopathogens in cattle in Beni Hamidene locality, district of Constantine (Νortheastern Algeria). Between June and October 2014, 169 bovines from 25 farms were included in this survey, 32 (18.9%) among them were suspected of piroplasmosis and/or anaplasmosis. Infection prevalences were estimated by microscopic examination of Giemsa‐stained blood smears and blood samples from all included cattle (n = 169). Animals were infected by Theileria annulata (65/169; 38.46%), Anaplasma marginale (22/169; 13%) and Babesia bovis (5/169; 3%). Two co‐infection patterns were found: Theileria annulata/Anaplasma marginale (7.69%) and Theileria annulata/Babesia bovis (1.18%). Only one farm had no cattle infected by any of the haemopathogens. There was a signification difference of T. annulata infection prevalence according to age category (p =.04). These results emphasised mainly the presence of bovine tropical theileriosis in northeastern, Beni Hamidene locality, province of Constantine, Algeria.

Molecular Characterization of Hemoparasites and Hemoplasmas Infecting Domestic Cats of Southern India.

In the present study, 111 blood samples were collected from apparently healthy cats belonging to four districts of Kerala, southern India, and they were investigated for the presence of hemoparasites and hemoplasmas by light microscopic examination and polymerase chain reaction (PCR). The microscopic examination of the Giemsa-stained blood smears did not reveal any parasites/organisms. However, PCR followed by nucleotide sequencing could detect 10 (9.01%) out of 111 samples infected with Hepatozoon felis, 3 (2.70%) with Cytauxzoon spp., and 10 (9.01%) with Mycoplasma spp. None of the samples revealed amplicons specific for the Babesia spp. and Trypanosoma evansi. The phylogenetic analysis of 18S ribosomal RNA (rRNA) gene sequences of H. felis revealed the existence of two different populations of H. felis circulating in the blood of infected cats. The phylogenetic tree was constructed based on 18S rRNA gene sequences of Cytauxzoon spp. and revealed that these isolates formed a distinct clade and do not cluster with any of the isolates from other countries. Among the 10 samples positive for Mycoplasma spp. infections, 7 were detected positive for Candidatus Mycoplasma haemominutum, two for Mycoplasma haemofelis, and one for Candidatus Mycoplasma turicensis. Phylogenetic analysis of 16S rRNA gene sequences of Mycoplasma spp. showed no distinct geographical grouping of the sequences. The sequences of M. haemofelis, Candidatus M. haemominutum, and Candidatus M. turicensis identified in the study clustered along with their respective isolates from around the world. To the best of our knowledge, this study forms the first report of molecular detection of Cytauxzoon spp. and Candidatus M. turicensis in cats from India.

Single Nucleotide Polymorphisms of Pfdhfr and Pfdhps Genes: Implications for Malaria Prophylactic Strategies in Maiduguri, Northeast Nigeria

Background. The success of Intermittent Preventive Treatment in Pregnancy (IPTp), Intermittent Preventive Treatment in Infancy (IPTi), and Seasonal Malaria Chemoprevention (SMC) depends on sulfadoxine-pyrimethamine (SP) efficacy. Objective. The study determined Single Nucleotide Polymorphisms (SNPs) of Plasmodium falciparum dihydrofolate reductase (Pfdhfr) and dihydropteroate synthase (Pfdhps) in Maiduguri, Northeast Nigeria. Materials and Methods. Giemsa-stained blood smears, capillary blood, and dried blood spot samples were collected from 63 subjects with uncomplicated malaria in Maiduguri between May and October 2018. Plasmodium species was determined and parasite density (PD) was estimated using the smears. Genomic DNA (gDNA) of P. falciparum was extracted from the dried blood spot samples using QIAamp DNA Mini Kit. The gDNA was subjected to nested PCR followed by restriction fragment length polymorphism (RFLP) to determine SNPs at Pfdhfr codons N51I, C59R, and S108N and Pfdhps codons S436A/F, A437G, and K540E. Results. The subjects’ mean age ± standard deviation was 23.6 ± 8.7 (2.0–67.0) years with a geometric mean PD of 8,948 (2,100–13,400) asexual parasites/µl blood. SNPs prevalence at any of the six Pfdhfr and Pfdhps codons was 85.7% (54/63); the prevalence was higher ( p &lt; 0.05 ) in Pfdhfr (82.5%; 52/63) than Pfdhps (58.7%; 37/63). Pfdhfr allele 108N (82.5%; 52/63) was the highest ( p &lt; 0.05 ) mutant when compared with alleles 51I (60.3%; 38/63) and 59R (66.7%; 42/63). Triple Pfdhfr mutation was observed in 60.3% (38/63) of the isolates and was higher ( p &lt; 0.05 ) among female subjects and SP recipients. Prevalence of Pfdhps allele 436A (28.6%; 18/63) was similar ( p &gt; 0.05 ) to allele 437G (34.9%; 22/63), with double mutation recorded in 4.8% (3/63). K540E mutation was not observed. Conclusion. Pfdhfr and Pfdhps mutations observed in Maiduguri are suggestive of SP resistance level, and this could constitute a setback to malaria prophylactic strategies in the region if unchecked. Thus, there is a need to investigate the clinical efficacy of SP.

Epidemiological link between canine monocytic ehrlichiosis caused by Ehrlichia canis and the presence of Rhipicephalus sanguineus sensu stricto in Argentina.

In this work, we analyze data that support an epidemiological link between cases of canine monocytic ehrlichiosis (CME) by Ehrlichia canis and the presence of Rhipicephalus sanguineus sensu stricto as vector in an endemic area for this tick in Argentina. In a blood sample of a 1-year-old toy poodle with CME compatible clinical signs, which showed CME typical morulae in monocytes in Giemsa-stained blood smear, DNA of E. canis was detected by PCR. Further, DNA of E. canis was also detected in a female of R. sanguineus s.s. collected on the infected dog. Rhipicephalus sanguineus s.s. is the only member of the R. sanguineus group that prevails in the study area. The results of this study suggest that R. sanguineus s.s. may play a more important role in the transmission of E. canis than it was assumed so far. The epidemiological link between CME cases and R. sanguineus s.s. as vector in temperate areas of Argentina described in this work contrast previous studies which found that R. sanguineus sensu lato "tropical lineage" (which is absent in the study area) is competent to transmit E. canis but not R. sanguineus s.s.

Long-lasting infectivity of Plasmodium vivax present in malarial patient blood to Anopheles aquasalis

Experimental studies for understanding the relationship between Plasmodium vivax and its vector hosts are difficult, because of to the lack of a long-term, in vitro continuous culture system unavailability of infected blood samples, seasonality of the disease, and the concentration of most cases in remote areas. This study evaluates the duration of the infectivity of P. vivax to Anopheles aquasalis after collecting blood from malaria-infected patients. Blood was collected from patients and stored at 4 °C and 37 °C. Every day, for 4 days, the blood was fed to An. aquasalis adult females, and a Giemsa-stained thick blood smear was mounted to account for sexual (gametocytes) and asexual (trophozoites and schizonts) stages and calculate parasitemia. Oocysts in the midgut of the mosquitoes were counted on the seventh day after feeding. Kruskal-Wallis test was used to compare the mean number of oocysts (MO) and the parasite density (PD) in each storage condition and post-infection time-points. The Mann-Whitney test was used to compare the number of oocysts for each day between temperatures. The results show that P. vivax stored at 4 °C and at 37 °C has its infectivity to An. aquasalis preserved for 2 days and 3 days, respectively. Infection rate (IR), PD and MO were higher on the day of blood collection and decreased gradually over time. The parasite density (number of parasites/μL) diminished faster at 4 °C than at 37 °C. In this study, a preservation protocol is shown for long-lasting infectivity of P. vivax in a blood sample taken from malaria-infected patients. These results show that infectivity of P. vivax stored at 4 °C and at 37 °C to An. aquasalis persist until 3 days after blood collection, but parasite density, infection rate, and mean of oocysts decreased 24h after blood collection. Since the malaria cases are increasingly far from the urban areas these results indicate that is possible, losing some infectivity, to realize experimental infections several dozen hours after the blood collection. However, it is necessary to improve the procedures for preserving P. vivax gametocytes for mosquito infection in the laboratory.

Deletions of pfhrp2 and pfhrp3 genes were uncommon in rapid diagnostic test-negative Plasmodium falciparum isolates from Uganda

BackgroundRapid diagnostic tests (RDTs) play a key role in malaria case management. The most widely used RDT identifies Plasmodium falciparum based on immunochromatographic recognition of P. falciparum histidine-rich protein 2 (PfHRP2). Deletion of the paralogous pfhrp2 and pfhrp3 genes leads to false-negative PfHRP2-based RDTs, and has been reported in P. falciparum infections from South America and Africa. However, identification of pfhrp2/pfhrp3 deletions has usually been based only on failure to amplify these genes using PCR, without confirmation based on PfHRP2 protein expression, and understanding of the true prevalence of deletions is incomplete.MethodsDeletions of pfhrp2/pfhrp3 in blood samples were investigated from cross-sectional surveys in 2012-13 in three regions of varied malaria transmission intensity in Uganda. Samples with positive Giemsa-stained thick blood smears, but negative PfHRP2-based RDTs were evaluated by PCR amplification of conserved subunit ribosomal DNA for Plasmodium species, PCR amplification of pfhrp2 and pfhrp3 genes to identify deletions, and bead-based immunoassays for expression of PfHRP2.ResultsOf 3516 samples collected in cross-sectional surveys, 1493 (42.5%) had positive blood smears, of which 96 (6.4%) were RDT-negative. Of these 96 RDT-negative samples, P. falciparum DNA was identified by PCR in 56 (58%) and only non-falciparum plasmodial DNA in 40 (42%). In all 56 P. falciparum-positive samples there was a failure to amplify pfhrp2 or pfhrp3: in 25 (45%) pfhrp2 was not amplified, in 39 (70%) pfhrp3 was not amplified, and in 19 (34%) neither gene was amplified. For the 39 P. falciparum-positive, RDT-negative samples available for analysis of protein expression, PfHRP2 was not identified by immunoassay in only four samples (10.3%); these four samples all had failure to amplify both pfhrp2 and pfhrp3 by PCR. Thus, only four of 96 (4.2%) smear-positive, RDT-negative samples had P. falciparum infections with deletion of pfhrp2 and pfhrp3 confirmed by failure to amplify the genes by PCR and lack of expression of PfHRP2 demonstrated by immunoassay.ConclusionFalse negative RDTs were uncommon. Deletions in pfhrp2 and pfhrp3 explained some of these false negatives, but most false negatives were not due to deletion of the pfhrp2 and pfhrp3 genes.

Bovine trypanosomosis: Prevalence and vector distribution in Sadi Chanka district, Kellem Wollega zone, Oromia regional state, Ethiopia

Bovine trypanosomosis remains an important livestock disease constraint, which is threatening livestock health and production, despite ongoing tsetse and trypanosomosis control efforts in Sadi Chanka district, Kellem Wollega zone, Oromia regional state, Ethiopia. A cross-sectional study was conducted in May 2018, to determine the prevalence of bovine trypanosomosis and distribution of the vectors of disease in Sadi Chanka district, Western Ethiopia. A total of 370 blood samples were collected from randomly selected local Horro and Abigar cattle breeds covering five villages of the district. The collected samples were examined using buffy coat microscopy and Giemsa-stained thin blood smear techniques. In this study, 12.4% (95% CI: 12.3-12.4) of the animals were found to be infected with trypanosomes. The study showed that 69.6% of trypanosome infections were caused by T. congolense followed by 26.1% T. vivax and 4.3% mixed T. congolense and T. vivax. In the present study, the association of bovine trypanosomosis was assessed in releation to body condition scores, sex, and age of cattle, and a significant association (P<0.05) was observed between body condition scores. However, significant differences were not observed between sex and age categories (P>0.05). The Mean Packed Cell Volume (PCV) of infected (21.6%) and non-infected (24.5%) groups of cattle had significant variation (P<0.05). In an entomological survey, a total of 616 flies were trapped, of which 280 (45.5%) were Glossina and the remaining 336 (54.5%) were Stomoxys, Tabanus, and Haematopota. The apparent density of Glossina, Stomoxys, Tabanus, and Haematopota was 3.5, 3.1, 0.7 and 0.4 fly per trap per day, respectively. This study generated basic scientific data on the epidemiology of bovine trypanosomosis and its vectors in Sadi Chanka district, which can be used in planning the control of bovine trypanosomosis in the area.

THE CIDR1α-PfEMP1 SEQUENCE FROM INDONESIAN PLASMODIUM FALCIPARUM AND ITS POTENTIAL ASSOCIATION WITH THE CEREBRAL OUTCOME

Background: Plasmodium falciparum Erythrocyte Membrane Protein 1 (PfEMP1) is an important protein responsible for the pathogenesis of severe malaria, including cerebral malaria. The protein is highly diverse. The CIDR1α-PfEMP1 binds endothelial protein receptor (EPCR) and may associated with the brain swelling in childhood malaria. Objective: To analyze the CIDR1α-PfEMP1 from Indonesian isolate and determine its association with cerebral malaria outcome. Methods: Fifteen blood samples of clinically mild to severe malaria-patient were collected for DNA extraction. Malaria diagnosis was conducted microscopically by Giemsa-stained thin blood smear. The CIDR1α domain was amplified by PCR using specific primer and PCR product was sequenced. The nucleotide sequences were analyzed by NCBI blast, DNASIS MAX 3 and translated into amino acid sequences using Expasy Translation Tool. Results: One out of fifteen samples was severe malaria case and infected with P. falciparum, the rest were clinically mild to moderate malaria and infected with pure P. falciparum or mixed infection of P. falciparum and P. vivax. Amplification for CIDR1α domain resulted a single band of + 550 bp from a severe sample only. Sequencing of PCR product on both strands read 524 nucleotides and BLAST analysis confirmed as CIDR1α sequence. Multiple alignment showed 74-78% nucleotide sequence similarity with reference sequences, but amino acid sequences presented 23.5% homologous. Conclusion: An identified CIDR1α domain only from severe case implicating the potential association with the severe outcome including cerebral malaria, but the highly diverse of the domain needs further studies on the interaction with the pathological-causing receptor in the host.

Co-infection of Babesia sp. and Theileria sp. in a Holstein Friesian crossbred calf- A case report

A 20-day-old Holstein Friesian, crossbred calf with a history of inappetence and weakness was presented to the veterinary hospital at Malhargarh, in recumbent condition. On clinical examination, the calf exhibited high fever, pale mucous membranes, dry muzzle, rough body coat and elevated heart and respiration rate. Giemsa stained blood smears revealed the presence of intra-erythrocytic Babesia and Theileria organisms. The calf was treated with diminazene aceturate, intramuscularly at a dose of 5 mg/kg body weight, along with supportive medication. However, the animal died after only a few hours of therapy. The present study reports the rare concurrent infection in a crossbred calf from Madhya Pradesh, India.

Haemotropic Mycoplasma sp. in a sow from Assam, India

Haemotropic Mycoplasma are specialized bacteria affecting red blood cells. They are associated with weakness, anaemia, stunted growth and mortality in piglets and reproductive failure in the adults. The present communication reports the detection of this organism (tentatively diagnosed as M. suis) in Assam in the blood of a pyrexic and recumbent sow brought for treatment at the Veterinary Clinical Complex of Lakhimpur College of Veterinary Science, Assam Agricultural University. Giemsa-stained blood smear revealed about 1% erythrocytes were infected with coccoid and ring forms of the organism. Estimation of red cell parameters in the blood sample revealed that haemoglobin (Hb), packed cell volume (PCV), total erythrocytes count (TEC) values were within the normal range. This is the first report of the organism from the state of Assam and further investigation on the extent of prevalence is warranted.

Anaplazmozisli Koyunlarda Bazı Inflamasyon Markırlarının Araştırılması

Anaplazmozis koyunlarda siklikla gorulen, kan hucrelerini enfekte eden artropod kaynakli ve ekonomik kayiplara neden olabilen paraziter bir hastaliktir. Bu calisma Anaplazmozis ile enfekte koyunlarda bazi serum inflamasyon markir seviyelerini degerlendirmek icin planlanmistir. Yapilan arastirma icin kullanilacak olan kan ornekleri, Van Buyuksehir Belediyesi Mezbahanesi’ne getirilen 20 saglikli ve 20 Anaplazmozis’li koyundan saglandi. Kontrol ve hasta gruplarinin belirlenmesi, hastaligin klinik belirtileri, giemsa boyali kan frotileri ve serolojik yontem (cELISA) yardimiyla yapildi. Toplanan 91 koyunlarin ait serum ornegi Anaplazma spp. turlerinin antikorlarinin varligi yonunde serolojik olarak incelendiginde %73.6’si (67/91) Anaplazmozis yonunden seropozitif bulundu. Kan frotisinde Anaplazma etkenleri gorunen ve ELISA inhibisyon degeri yuksek cikan koyunlarin kanlari analizler icin kullanildi. Anaplazmozis’li koyunlar teshis edildikten sonra pozitif ve negatif olarak gruplandirildi. Pozitifligi tespit edilen 20 koyun serumunda granulosit makrofaj koloni uyarici faktor (GM-CSF), interloykin-1β (IL-1β), interloykin-6 (IL-6), tumor nekroz faktor- alfa (TNF-α) duzeyleri elisa kiti ile kit prosedurune gore belirlendi. Proinflamasyon parametrelerinden serum IL-1β (ng/L), TNF-α (ng/L), GM-CSF (ng/L) duzeylerinin negatif gruba gore istatiksel olarak yukseldigi tespit edildi. Anaplazmaya karsi gelisen inflamasyon ile ilgili mekanizmalarin incelenmesi, ileride gelistirilecek immunoterapi arastirmalari icin Anaplazma enfeksiyonlarinin teshis ve onlemeye yonelik calismalara katki saglayacagi dusunulmektedir.