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Equine piroplasmosis is a worldwide tick-borne disease caused by the parasites Theileria equi sensu lato and Babesia caballi, with significant economic and sanitary consequences. These two parasites are genetically variable, with a potential impact on diagnostic accuracy. Our study aimed to evaluate the frequency of asymptomatic carriers of these parasites in France and describe the circulating genotypes. We developed a species-specific nested PCR protocol targeting the 18S small sub-unit (SSU) rRNA gene and used it on blood samples collected from 566 asymptomatic horses across four National Veterinary Schools. The carrier frequency varied from 18.7 % around Paris (central-north) to 56.1 % around Lyon (southeast), with an overall prevalence of 38.3 %. Theileria equi carriers were ten times more frequent (91.7 %; 209/228 isolates) compared to B. caballi carriers (8.3 %; 19/228 isolates). Notably, T. equi carrier frequency was significantly lower in the northern region (Ile de France) compared to the southeastern regions. A positive relationship was observed between the frequencies of asymptomatic carriers and the frequency of previous acute piroplasmosis reported from the owner across all four geographic areas. Neither horse gender nor age showed a significant effect on the frequency of asymptomatic carriers. In some areas, a substantial proportion of horses (22.2 % to 37.5 %) carried T. equi before the age of three years, indicating high infection pressure. Genotyping of 201 T. equi isolates revealed a predominance of genotype E (98 %) and few isolates belonging to genotype A (2 %). Notably, two of the four T. equi genotype A isolates were detected in horses originating from Spain. All 19 B. caballi isolates belonged to the genotype A. The discussion section explores the link between these results, the tick distribution and abundance, and the frequency of detection of T. equi and B. caballi in febrile cases attributed to piroplasmosis., Competing Interests: Declaration of competing interest The authors declare that they have no competing interests., (Copyright © 2024. Published by Elsevier GmbH.)

We report an unusual case of strongyloidiasis in a 62-year-old male, presenting with fever, and acute diarrhea. The patients had concomitant bilateral renal parenchymal disease and carrier for Hepatitis B. Numerous motile larvae were observed in wet mount of the freshly passed stool. The patient responded well to oral ivermectin. Prompt and accurate diagnosis of strongyloidiasis can prevent the consequences associated with hyperinfective syndrome., Competing Interests: Declaration of competing interest Nil., (Copyright © 2024 Indian Association of Medical Microbiologists. Published by Elsevier B.V. All rights reserved.)

Equine piroplasmosis (EP) is caused by Theileria equi and Babesia caballi, transmitted by tick vectors. Horses can suffer an acute, subacute, and chronic forms of the disease, with clinical signs such as poor performance, fever, pale mucosal membranes, and jaundice. The diagnosis of EP subclinical cases is complex due to the sensitivity of real-time PCR and the limited parasite load in some carriers, making it challenging to differentiate them from seropositive, PCR negative (S+PCR-) individuals. This study aimed to describe haematological and biochemical changes in asymptomatic EP carriers, EP S+PCR- horses and control horses (EP seronegative and PCR negative). It also investigated potential haemato-biochemical markers to aid in distinguishing true EP carriers alongside molecular and serological tests. A comprehensive haematology and biochemistry profile was conducted on 410 sera and EDTA blood samples, comprising 130 EP positives by real-time PCR and competitive ELISA (cELISA) (carriers), 130 EP negatives by real-time PCR but positive to cELISA (S+PCR-) and 150 EP negative horses to real-time PCR and c-ELISA (controls). Our study confirmed that a haematological and biochemistry profile could help to differentiate between EP carriers/S+PCR- from healthy horses. Carriers and S+PCR- horses showed significant increases in the white blood cell count (WBC), high total proteins (TP) and total globulins (GLOB) concentration, and liver function markers compared to controls. Additionally, the evaluation of uric acid (UA) suggested oxidative stress in carrier horses. However, no useful haemato-biochemical diagnostic markers were identified to aid the challenging differentiation of EP carriers and S+PCR- horses, highlighting the need for improvement in molecular/serological diagnosis for these horses., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests:Eliazar Camino reports financial support was provided by Complutense University of Madrid. Eliazar Camino reports a relationship with Complutense University of Madrid that includes: funding grants., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

The tick-borne equine hemoparasite, Theileria equi, is endemic in many parts of the world where prevalence may be high, and most infected horses are apparently healthy but serve as life-long carriers. To determine the factors that affect T. equi dynamics, we followed parasitic loads in apparently healthy horses at four time points during one year. A total of 1094 blood samples were collected from 395 horses, along with ticks and demographic and clinical data. Infection and load of T. equi were tested by PCR and qPCR, and for the spring dataset, infection was also tested serologically by IFAT (n = 268). Theileria equi was molecularly detected in 64.8 % of the horses. The agreement between molecular and serological results was 79.8 % (K > 0.674) and positively correlated with parasitic load. Infection was associated with pale mucus membranes, lower packed cell volume and higher total solids (all P < 0.001), although these changes had only minor clinical importance. While parasitic loads in qPCR-positive samples (n = 561) were generally low (mean = 7.9-10 -4 % parasitized erythrocytes), younger horses showed higher loads, possibly suggesting development of immunity. Infection and parasitic load were associated with housing management and tick exposure, illustrating different patterns of exposure. Endemic stability is suggested in pasture farms with constant exposure to ticks, where parasite prevalence was high (96 %) and associated with T. equi 18S rRNA genotype D, low parasitemia and high antibody titers. Endemic instability can be suggested in case were horses are kept in paddocks (prevalence = 49 %) with intermittent exposure to ticks, where infection was associated with high parasitemia when ticks were present. A steady state is suggested in stabled horses (prevalence = 46 %), with no exposure to ticks; where infection was associated with genotype A, low parasitemia and low antibody titers. The ability to identify different risk groups within endemic areas may improve the administration of suitable treatment and control practices in an effort to reduce the risk of clinical disease., (Copyright © 2020 Elsevier GmbH. All rights reserved.)

Asymptomatic outdoor dogs can be carriers of Babesia canis, but data describing the development of an acute phase response (APR) are not available. We hypothesised that these dogs have a moderate APR that could be detected by hematological and biochemical changes. Two groups of Babesia-exposed dogs were represented by nine B. canis PCR-positive and twenty B. canis PCR-negative, seroreactive dogs. The control group consisted of ten Babesia-naïve dogs. Serum amyloid A (SAA), paraoxonase-1 (PON-1), complete blood count, and biochemistry parameters were analysed by standard methodologies. Protein and lipoprotein fractions were separated using agarose gel electrophoresis (GE), and the dominant diameters of lipoproteins were assessed on gradient GE. Results were evaluated using non-parametric tests and the Receiver Operating Characteristic curve. SAA (median 39.0 μg/mL, range 2.2-48.8 μg/mL), total protein (median 74.7 g/L, range 57.1-98.3 g/L) and the dominant diameter of α-lipoproteins (median 13.31 nm, range 12.09-14.17 nm) in B. canis PCR-positive dogs were higher relative to dogs in the control group or dogs that were PCR-negative but seroreactive (p < 0.001 for both groups). Mild to moderate anemia (4/29), thrombocytopenia (7/29), and leukocyte counts that were close to the upper limit of the reference range were encountered in both Babesia-exposed groups. When compared to controls, Babesia-exposed dogs displayed decreased a PON-1 activity and protein GE pattern consistent with low-grade chronic inflammation (p < 0.001 for both groups). Dogs with detectable amounts of B. canis DNA in blood contain increased levels of SAA and total protein along with α-lipoproteins that display an increased diameter relative to those dogs with positive Babesia serology but undetectable levels of B. canis DNA in blood., (Copyright © 2020 Elsevier B.V. All rights reserved.)

East Coast fever (Theileria parva infection in cattle) was eradicated from South Africa in the mid-1900. However, another form named Corridor disease (CD), associated with T. parva carrier buffaloes exists and outbreaks have increased in endemic areas. The occurrence of a CD carrier state in cattle under field situations has not been demonstrated but remains a subject of controversy. The current study investigated the T. parva carrier state following a severe outbreak in cattle introduced onto a game ranch. Monitoring of the outbreak included clinical signs, mortality, microscopy, serology, real-time PCR and xenodiagnoses. The herd of cattle received block treatment using oxytetracyclines (OTC) by the farmer during the outbreak. Cattle were sampled early during the outbreak and twice within the following 75 days. All buffaloes were tested for a T. parva carrier state. Two batches of questing adult R. appendiculatus were collected at the time of disease occurrence and a year later. These ticks were fed on susceptible cattle under controlled conditions and monitored for disease transmission. Ticks infected with a buffalo-derived stock of T. parva were fed on one bovine under controlled conditions and simultaneously injected with OTC, simulating the infection and treatment method of vaccination and was used as a positive control. Clean R. appendiculatus nymphs were fed on four recovered PCR positive cattle from the outbreak and on the positive control animal. The adult ticks were tested for infectivity by xenodiagnoses on susceptible bovines. For the initial outbreak the CD prevalence was 62.3% with a mortality rate of 29.5%. However, the outbreak was contained by block OTC treatment of the herd since only 3.4% cattle subsequently died until the end of the investigations. Adult ticks fed on one field bovine and the laboratory established T. parva carrier both transmitted fatal infections to susceptible cattle. Ticks fed on two field cattle transmitted T. taurotragi and one failed to transmit any infection. Questing adult R. appendiculatus collected during the outbreak transmitted fatal CD to two bovines while ticks collected a year later transmitted T. taurotragi. These findings demonstrated the effectiveness of disease control either by cattle treatment using OTC simulating the ITM or by intensive cattle dipping following the outbreak or by both interventions. The potential risk of creating carrier cattle by OTC treatment during CD outbreaks should be considered, supporting the continued control measures of segregation of cattle and buffalo herds., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Infection and Treatment Method (ITM) has been practiced in Tanzania for over 20 years as a prevention measure against East Coast Fever disease. It is known that ITM, like natural ECF infection, leads to a carrier state, whereby vaccinated cattle become asymptomatic carriers of the parasite. It is expected that ECF vaccination using ITM also leads to generation of combinations of vaccine specific Theileria parva and local strains that circulate in the field what contributes to an unknown level of parasite diversity. Moreover, the long term impact of ITM on carrier state and parasite diversity in cattle are largely unknown. To address this question blood was collected from ECF-vaccinated (n = 239) and unvaccinated (n = 97) cattle from Loiborsoit, Emboreet, Esilalei, Manyara ranch and Mswakini villages in the Maasai steppe of northern Tanzania, as well as Mruazi and Leila farms in Tanga in eastern Tanzania. Screening for T. parva using nested PCR revealed an overall prevalence of T. parva to be 34.5%, with a significant higher prevalence among ECF-vaccinated cattle. Using three VNTR markers (ms2, ms5 and MS7) higher parasite genetic diversity in terms of higher number of alleles and expected heterozygosity was shown in vaccinated than unvaccinated cattle. These parameters were highest in cattle from Manyara ranch. Nevertheless, the principle component analysis (PCoA) showed no distinct clustering patterns as most T. parva alleles clustered together throughout the four quadrants implying parasite homogeneity among the sampled populations. However, some of the parasite alleles closely clustered with Muguga vaccine alleles in two of the quadrants, consistent with closer genetic relatedness between the vaccine strains and the T. parva populations from the Maasai steppe. Likewise analysis of molecular variance (AMOVA) revealed most of the genetic variation (93%) being contained within populations with only 7% being among populations. This study therefore confirms the role of ECF vaccination in enhancing carrier state and T. parva diversity in vaccinated cattle populations. Higher T. parva diversity may play an important role in carrier cattle by way of restricting breakthrough infections from field parasite strains., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Baylisascaris procyonis, the roundworm of raccoons (Procyon lotor), is an emerging helminthic zoonosis in North America. Since the larval form is capable of causing neurological disease in more than 150 species of birds and mammals including humans, understanding factors that influence carriage of the parasite by raccoons is important for mitigating risk. This review examines the current literature to identify major demographic and environmental risk factors associated with B. procyonis carriage in wild raccoons. Raccoon age and season of sample collection were most commonly identified as risk factors, with increased prevalence found in juvenile animals and when sample collection occurred in the fall. Human urbanization and agricultural land use were also observed as potential risk factors; however, there are inconsistencies in the direction of influence these risk factors have on the prevalence of infection. Further investigation into the role of environmental risk factors is required to better understand how human activities influence parasite carriage in raccoons. Additionally, future research using multivariable statistical models guided by epidemiological principles to control for confounding variables and identify interaction effects will help clarify the effect of these demographic and environmental factors. Developing a better understanding of the primary risk factors for parasite carriage in raccoons will help identify areas of higher risk for environmental contamination and will aid in the development and refinement of education and management programs to reduce the risk of human exposure., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Across the Americas and the Caribbean, nearly 561,000 slide-confirmed malaria infections were reported officially in 2008. The nine Amazonian countries accounted for 89% of these infections; Brazil and Peru alone contributed 56% and 7% of them, respectively. Local populations of the relatively neglected parasite Plasmodium vivax, which currently accounts for 77% of the regional malaria burden, are extremely diverse genetically and geographically structured. At a time when malaria elimination is placed on the public health agenda of several endemic countries, it remains unclear why malaria proved so difficult to control in areas of relatively low levels of transmission such as the Amazon Basin. We hypothesize that asymptomatic parasite carriage and massive environmental changes that affect vector abundance and behavior are major contributors to malaria transmission in epidemiologically diverse areas across the Amazon Basin. Here we review available data supporting this hypothesis and discuss their implications for current and future malaria intervention policies in the region. Given that locally generated scientific evidence is urgently required to support malaria control interventions in Amazonia, we briefly describe the aims of our current field-oriented malaria research in rural villages and gold-mining enclaves in Peru and a recently opened agricultural settlement in Brazil., (Copyright © 2011 Elsevier B.V. All rights reserved.)

At a time when human African trypanosomiasis (HAT) elimination again seems a reachable goal in many parts of sub-Saharan Africa, it is becoming increasingly important to characterise the factors involved in disease resurgence or maintenance to develop sustainable control strategies. In this study conducted in the Forecariah mangrove focus in Guinea, HAT patients and serological suspects (SERO) were identified through mass screening of the population with the Card Agglutination Test for Trypanosomiasis (CATT) and were followed up for up to 2 years. Analysis of the samples collected during the follow-up of HAT patients and SERO was performed with PCR (TBR1/TBR2) and the trypanolysis serological test (TL) in order to clarify the role played by these individuals in the epidemiology of HAT. PCR positivity was higher in TL⁺ than in SERO TL⁻ (50% vs. 18%, respectively). Whereas CATT plasma titres decreased both in treated HAT patients and SERO TL⁻, SERO TL⁺ maintained high CATT titres. Four out of 17 SERO TL⁺ developed HAT during the study. These results strongly suggest that SERO TL⁺ individuals are asymptomatic carriers. In the context where disease prevalence is sufficiently low, treating SERO TL⁺ individual may thus be of crucial importance in order to cut transmission., (Copyright © 2011 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.)

Serologic and molecular evidence suggest that white-tailed deer in South Texas and North Mexico carry the agents of bovine babesiosis, Babesia bovis and Babesia bigemina. To determine if white-tailed deer in central Texas, which is outside the known occurrence of the vector tick at this time, harbor these parasites, blood samples from free-ranging and captive white-tailed deer (Odocoileus virginianus) in Tom Green County were tested by polymerase chain reaction (PCR) assays for B. bovis and B. bigemina 18S rDNA. Of the 25 samples tested, three (12%) were positive by nested PCR for B. bovis. This identity was confirmed by sequence analysis of the cloned 18S rDNA PCR product. Further confirmation was made by sequence analysis of the rRNA internal transcribed spacer (ITS) 1, 5.8S rRNA gene, and ITS 2 genomic region in two (representing samples from two different ranches) of the B. bovis positive samples. Three samples were positive by B. bigemina nested PCR, but sequencing of the cloned products confirmed only one animal positive for B. bigemina; Theileria spp. DNA was amplified from the other two animal samples. In addition to Theileria spp., two genotypically unique Babesia species sequences were identified among the cloned sequences produced by the B. bigemina primers in one sample. Phylogenetic analysis showed no separation of the deer B. bovis or B. bigemina 18S rDNA, or deer B. bovis ITS region sequences from those of bovine origin. Clarification of the possible role of white-tailed deer as reservoir hosts in maintaining these important pathogens of cattle is critical to understanding whether or not deer contribute to the epidemiology of bovine babesiosis., (Copyright © 2010. Published by Elsevier B.V.)

Cape buffalo (Syncerus caffer) are considered to be an important reservoir for various tick-borne haemoparasites of veterinary importance. In this study we have compared the haemoparasite carrier prevalence in buffalo from four geographically isolated national parks in Uganda [Lake Mburo National Park (LMNP), Queen Elizabeth National Park (QENP), Murchison Falls National Park (MFNP) and Kidepo Valley National Park (KVNP)]. Differences were seen in haemoparasite prevalence in buffalo from the four national parks. All the buffalo sampled in LMNP were carriers of Theileria parva however, buffalo from MFNP and KVNP, which are both located in the north of Uganda, were negative for T. parva. Interestingly, 95% of buffalo in the northern part of QENP were T. parva positive, however all buffalo sampled in the south of the park were negative. A high multiplicity of infection was recorded in all the buffalo found to be carrying T. parva, with evidence of at least nine parasite genotypes in some animals. Most of the buffalo sampled in all four national parks were carriers of T. mutans and T. velifera, however none were carriers of T. taurotragi, Babesia bovis, Babesia bigemina, Ehrlichia bovis or Ehrlichia ruminantium. All the buffalo sampled from LMNP were positive for T. buffeli and T. sp. (buffalo) however, buffalo from the parks in the north of the country (KVNP and MFNP) were negative for these haemoparasites. Anaplasma centrale and Anaplasma marginale were circulating in buffalo from all four national parks. T. parva gene pools from two geographically separated populations of buffalo in two of the national parks in Uganda (LMNP and QENP) were compared. The T. parva populations in the two national parks were distinct, indicating that there was limited gene flow between the populations. The results presented highlight the complexity of tick-borne pathogen infections in buffalo and the significant role that buffalo may play as reservoir hosts for veterinary haemoparasites that have the potential to cause severe disease in domestic cattle., (Crown Copyright © 2010. Published by Elsevier B.V. All rights reserved.)

Unlabelled: Cystic echinococcosis is an endemic disease in the Province of Rio Negro, Argentina. Ultrasound surveys carried out in 1984 found prevalence rates of 5.6% in children between 6 and 14 years of age., Objective: To describe and to evaluate the results of the strategy applied in school children by hospital services of the Province of Rio Negro with regard to diagnosis, treatment and monitoring of cystic echinococcosis and to evaluate simultaneously the results of the control program against cystic echinococcosis., Materials and Methods: In 1997 ultrasound was chosen to carry out population surveys and the medical treatment criteria for the detected cases were standardized. The population under study involved 5745 students in the first survey and 22,793 in subsequent studies. The detected cases were classified according to Gharbi's scheme. A treatment algorithm was defined based only on monitoring ("watch and wait"), albendazole, surgery (open or laparoscopic) or mini-invasive procedures, according to type, location and size of the cyst. Information was also obtained on cases notified to the Health System between 1980 and 2008., Results: In the first survey, 70 carriers (1.2%) were detected; of these, 25 started albendazole treatment (35.7%) and only 3 (4.3%) underwent surgery. Ten years after treatment, 60.1% of 42 cases, presented Types IV and V cysts and 14.5% presented total involution of their cysts. In subsequent studies, 87 (0.4%) cases were detected, 49 of which started albendazole treatment (56.3%) and 9 underwent surgery (10.3%). The incidence rate of cystic echinococcosis cases decreased from 38×100,000 in 1980 to 3.7×100,000 in 2008., Discussion: A strong decrease in cystic echinococcosis was obtained although persistent levels of transmission were maintained. The cases produced under these conditions are diagnosed by means of ultrasound surveys and are treated using a plan based on albendazole and monitoring by the Health System during a period of 10 years., (Copyright © 2010 Elsevier B.V. All rights reserved.)

Besnoitia besnoiti, an apicomplexan parasite causes economically important disease in cattle in many countries of Africa and Asia is re-emerging in Europe. Serological identification of infected cattle is important because introduction of these animals into naive herds seems to play a major role in the transmission of the parasite. We report new, simplified immunoblot-based serological tests for the detection of B. besnoiti-specific antibodies. Antigens were used under non-reducing conditions in the immunoblots, because reduction of the antigen with beta-mercaptoethanol diminished the antigenicity in both, tachyzoites and bradyzoites. Ten B. besnoiti tachyzoite and ten bradyzoite antigens of 15-45 kDa molecular weight were recognized by B. besnoiti infected cattle, but not or only weakly detected by cattle infected with related protozoan parasites, Neospora caninum, Toxoplasma gondii, Sarcocystis cruzi, Sarcocystis hominis, or Sarcocystis hirsuta. The sensitivity and specificity of B. besnoiti immunoblots were determined with sera from 62 German cattle with clinically confirmed besnoitiosis and 404 sera from unexposed German cattle including 214 sera from animals with a N. caninum-specific antibody response. Using a new scoring system, the highest specificity (100%) and sensitivity (90%) of the immunoblots were observed when reactivity to at least four of the ten selected tachyzoite or bradyzoite antigens was considered as positive. When a cut-off based on this scoring system was applied to both the tachyzoite- and the bradyzoite-based immunoblots, there was an almost perfect agreement with the indirect fluorescent antibody test with a titre of 200 as the positive cut-off. We identified and partially characterized 10 tachyzoite and 10 bradyzoite B. besnoiti antigens which may help to develop new specific and sensitive serological tests based on individual antigens and in the identification of possible vaccine candidates., (Copyright (c) 2010 Elsevier B.V. All rights reserved.)

Trypanosoma evansi, the causative organism of 'surra' expresses its variable surface glycoprotein (VSG) at early, middle and late stages of infection in animals. The variable antigenic nature of VSG caused by switching its expression type favours evasion from the host immune response and leads to chronic and persistent infection. Developing a polymerase chain reaction (PCR)-based diagnostic tool targeting the VSG gene is expected to be highly specific and sensitive for diagnosis of surra. Hence, in the present study, we have designed EXP3F/4R primer pair and amplified the 1.4 kb of VSG gene of T. evansi and studied the phylogenetic relationship by in silico analysis. The PCR method was standardised using another set of primer, DITRYF/R, and 400 bp was amplified from blood and tissue samples of experimentally infected animals. Applying the PCR method, we were able to detect as low as 0.15 trypanosomeml(-1). Considering the number of parasite-to-DNA concentration, the PCR method has a sensitivity of 0.015 pg ml(-1). The PCR could detect the presence of the parasite as early as 24h post-infection (p.i.) and 72 h p.i., respectively, in experimentally infected rats and buffalo. No amplification was observed with DNA of Babesia bigemina and Theileria annulata, indicating the primers are specific for T. evansi. The PCR method could detect the dog, lion and leopard isolates of T. evansi. Similarly, amplifying the DNA from the experimentally infected tissues was also found to be sensitive. Thus, the findings of this study favour the application of PCR over the parasitological methods for the detection of the early and/or chronic stage of surra in domestic and wild animals., (Copyright (c) 2010 Elsevier B.V. All rights reserved.)

Entamoeba moshkovskii and Entamoeba dispar are microscopically indistinguishable from the pathogenic species Entamoeba histolytica. There are limited data on the prevalence of these commensal infections from Africa. We utilized PCR and antigen detection to evaluate the carriage rate of E. moshkovskii, E. dispar, and E. histolytica infection in stool from a cohort of HIV-suspected or confirmed inpatients from Tanzania. E. histolytica was detected by ELISA in 4% (5/118) while E. moshkovskii and E. dispar were detected by PCR in 13% (18/136) and 5% (7/136) of individuals, respectively (P<0.05). Supporting their commensal nature, neither E. moshkovskii nor E. dispar infection was statistically associated with HIV status, CD4 count, or the presence of diarrhea. These data suggest E. moshkovskii is a common infection in HIV-infected individuals in northern Tanzania and supports the concept that the microscopic detection of Entamoeba should be interpreted cautiously.

Congo-Brazzaville has recently adopted artesunate-amodiaquine as the first-line antimalarial drug to replace chloroquine. Before the implementation of this new strategy, we conducted several clinical studies to assess the therapeutic efficacy of former, classical first-line antimalarial drugs in the city of Brazzaville, in which reside about 30% of the Congolese population. From 2003 to 2005, non-randomised trials were conducted to evaluate the efficacy of sulfadoxine-pyrimethamine (SP) (n=97 patients), amodiaquine (AQ) (n=62 patients), and the combination of sulfadoxine-pyrimethamine-amodiaquine (n=54 patients) in children aged between 6 months and 5 years with uncomplicated malaria using the 2003 WHO guidelines during the 28-day follow-up period. After excluding new infections by PCR, the proportion of treatment failure on day 28 was 30.2% (95% confidence interval, 19.2-43.0%) for sulfadoxine-pyrimethamine, 34.8% (95% confidence interval, 21.4-50.2%) for amodiaquine, and 14.2% (95% confidence interval, 5.9-27.2%) for sulfadoxine-pyrimethamine+amodiaquine combination. Treatment with sulfadoxine-pyrimethamine was associated with an increase of gametocyte charge. These results suggest that neither sulfadoxine-pyrimethamine nor amodiaquine is efficacious as monotherapy and that their combination may not remain effective in the coming years. Based on our results, the implementation of artemisinin-based combination therapy appears to be urgent in the country.

Reduction of transmission is important for effective malaria control. Standard microscopic detection grossly underestimates the human transmission reservoir. Malaria transmission intensity is defined by local and focal conditions. Transmission-blocking vaccines may be effective by directly targeting identified hot spots of transmission.

The present study sought to elucidate whether Schistosoma japonicum eggs could survive a passage through dogs and chickens. Three dogs and three chickens were used for the experiment. The dogs were allowed to eat goat faeces containing 80 S. japonicum eggs per gram. The chickens were given 2 ml of water containing more than 100 S. japonicum eggs per ml. The hatchability of the eggs was 51.8% prior to administration. All faeces were collected for 3 days from the dogs and for 2 days from the chickens and egg excretion and hatchability were measured. The results showed that nearly 50% of the eggs administered to the dogs were recovered in the faeces during the first 2 days and during the first 25 h 39% of the recovered eggs were viable and able to hatch. In the chickens 9% of the administered eggs were recovered within 27 h and within the first 3.5 h after administration 19% of the recovered eggs were viable and able to hatch. The results suggest that both dogs and chickens may act as transport hosts for S. japonicum in endemic areas.

The existence of a pig reservoir for human African trypanosomosis (HAT) due to Trypanosoma brucei gambiense complicates the fight against this disease. This study, reports results obtained from pigs, which were inoculated with the blood of a person, suffering from HAT in Cameroon. The pigs were reared and kept in the shelter from all contact with Glossina, and monitored for 188 days. The seroconversion was checked by agglutination assays for trypanosomosis (CATT 1.3 and LATEX/T.b.gambiense). The parasitemia was measured by quantitative buffy coat method (QBC) and by polymerase chain reaction method (PCR). In addition, growth was recorded as well as blood counting and blood formulas. The results showed that the pigs were trypanotolerant and cure themselves in less than 6 months. It is concluded that sterilisation of this reservoir could be achieved by tsetse-control measures in 1 year. It confirms the strategy to complement screening and treatment of HAT with tsetse fly control measures.

Outbreaks of autochthonous babesiosis, caused by Babesia canis, occurred in The Netherlands in the spring and autumn of 2004 affecting 23 dogs. Nineteen animals recovered after treatment, whereas four dogs died. Adult Dermacentor reticulatus ticks collected from these dogs indicate that canine babesiosis could become endemic in The Netherlands.

The avidity (functional affinity) of specific antibodies are being used to estimate duration of bovine Neospora caninum infection. Here, we report for the first time the avidity pattern in cattle orally inoculated with N. caninum oocysts. In all, 16 pregnant cows and 7 calves were administered N. caninum oocysts. In the cows, the avidity increased during the early course of infection. In all but one, the avidity was < or = 35 during the first 6 weeks after infection and no cow had an avidity value >50 until week 9. The calves were sampled either week 6 (n = 3) or week 9 (n = 9) after infection, and by then had avidities between 2 and 17. The results are in agreement with results from previous investigations of naturally infected cattle, and calves that were experimentally infected with tachyzoites. They further validate the ability of the N. caninum iscom avidity ELISA to accurately assess the duration of infection.

Aim: To determine the contribution of Neospora caninum to abortions on a dairy farm in NSW (Australia), determine the mode of transmission and develop and trial a control option for infection., Methods: Two whole herd bleeds were conducted 12 months apart and the association between serological status and abortion events were calculated for a number of bovine abortifacients. Family trees were constructed for N. caninum seropositive cattle in the herd. Some N. caninum seropositive cows were culled from the herd and no female offspring was retained from seropositive cows., Results: At the first whole-herd bleed in December 2002 a seroprevalence of 10.2% for N. caninum infection was detected. Cows with N. caninum infection were 13 times more likely to abort than uninfected ones. Seventy-five percent of seropositive animals in the herd were related, suggesting a high degree of congenital infection/transmission. Only 15% of infections were likely to be postnatally acquired. Selective culling of seropositive cows and not breeding from them reduced the number of seropositive animals. Only one newly sero-converted cow was detected at the second whole-herd bleed 12 months later., Conclusions: Seroepidemiological approaches were able to establish a high degree of association between N. caninum infection and low-level abortion in the dairy herd. Vertical transmission of infection was the predominant mode of infection and hence control efforts aimed at selectively culling seropositive animals from the herd were highly successful in reducing the level of infection.

Anaplasmosis, caused by the tick-borne rickettsia, Anaplasma marginale, is an economically important disease of cattle in the United States and worldwide. Cattle that recover from acute infection become carriers in which low or microscopically undetectable A. marginale rickettsemia persists. Tetracycline antimicrobials are currently the only drug used in the US for treatment of acute anaplasmosis. There are currently no drugs specifically licensed for elimination of persistent infections. This study tested the efficacy of three oxytetracycline treatment regimens to clear A. marginale from cattle that were persistently infected. Forty Angus x Simmental steers, aged 6-12 months were experimentally infected with A. marginale. After the steers recovered from acute infection, seroconverted, and were confirmed infected using nested PCR followed by DNA hybridization, the carrier status of each animal was ascertained by sub-inoculation of blood into a separate, splenectomized Holstein calf. The steers were then blocked by bodyweight and randomly assigned as follows to four treatment groups: Treatment A, 300 mg/ml solution of oxytetracycline (Tetradure LA-300, Merial Canada Inc.) administered at 30 mg/kg, by intramuscular (i.m.) injection on day 0; Treatment B, the same 300 mg/ml solution of oxytetracycline administered at 30 mg/kg, i.m. on day 0 and again on day 5; Treatment C, a 200 mg/ml solution of oxytetracycline (Liquamycin LA-200, Pfizer Animal Health) administered at 22 mg/kg, intravenously (i.v.), q 24 h for 5 days (a treatment dose that corresponds with current Office International des Epizooties (OIE) recommendations for treatment prior to export). The fourth group consisted of untreated infected control cattle. All steers were still nested PCR and cELISA positive at 60 days after treatment. Infection was confirmed by subinoculation of blood into a splenectomized Holstein calf. These results demonstrated that the treatment regimens tested failed to clear A. marginale infections in carrier cattle.

To assess the epidemiology of Babesia divergens in a veterinary practice based in the mid-east of France ("Monts du Lyonnais"), blood was collected from 254 cattle belonging to 24 herds. To assess the dynamics of the carrier state, six carriers were identified, treated with flumethrin and sampled once every 3 weeks during 6 months. Two different DNA extraction methods were compared. Each sample was tested for the presence of parasites using a PCR-RFLP test based on the 18S rRNA gene. The sensitivity of the test was equivalent to a parasitaemia as low as 10(-5)% (in "Filter Paper" samples) and 10(-6)% in 1 ml blood (extracted using "Matrix"). With the latter method, the rate of detection diminishes in the low parasitaemia range but could probably be improved. This test proved to be very useful in the detection of B. divergens carriers. Serology using IFAT showed 7% of the cattle seropositive, which is suggestive of a disease situation with a low clinical risk level. Analysis of the PCR results suggests a 20% prevalence rate of carriers in the cattle population. The use of the mean parasitaemia is proposed to serve as a babesiosis clinical risk indicator. This approach could also be used in other babesia infections provided the lowest detectable parasitaemia level (threshold level) could be resolved for each parasite species.

Hydatidosis or cystic echinococcosis (CE) caused by Echinococcus granulosus is endemic in the Province of Río Negro, Argentina. The objective of this investigation was to evaluate the results of a program carried out in endemic areas of the Province of Río Negro, Argentina, in the years 1997-2002. Abdominal ultrasonography was used, classifying the cases detected according to WHO guidelines. A treatment algorithm was defined which included observation, albendazol therapy, PAIR or surgery, according to cyst type and size. A total of 5745 schoolchildren were evaluated, detecting hydatid cyst carriers in 70 (1.2%). Of these; 40 (57.1%) were included in follow-up protocol, 25 (35.7%) in treatment protocol with albendazol, 2 (2.9%) with PAIR and 3 (4.3%) with conventional surgery. After a mean of 44 months, among 25 cases treated with albendazol, in 2 (8%) cysts underwent total involution, in 17 (68%) they presented positive changes, in one (4%) they remained unchanged and in 4 (16%) they progressed to type II, while 1 (4%) displayed negative evolutionary changes. Out of 39 cases under observation alone protocol, in 8 cases (21%) cysts underwent total involution, in 7 (18%) they presented positive changes, in 11 (28%) they remained unchanged, in 2 (5%) they progressed to Type II and in 11 (28%) they presented negative evolutionary changes and had to be included in the other protocol types. In this study, conventional surgery, was applied to 10% of detected cases. The combination of ultrasonographic screening and albendazol treatment showed promising results.

The course of Trypanosoma evansi infection in coatis (Carnivora, Procionidae) was followed for 262 days. Parasites were detected in all infected animals from day 2 post infection until the end of the study. No correlation between temperature and parasitemia was observed. Animals of the infected group demonstrated depression, weakness, lethargy and pale mucous membranes. Indirect fluorescent antibody tests detected anti-T. evansi antibodies within 7 to 14 days post infection and showed high levels until the end of the experimental period. The persistent parasitemia in coati and their relative tolerance to clinical signs suggested that this species develops a chronic disease and plays an important role in the epidemiology of trypanosomosis due to T. evansi in enzootic regions.

Entamoeba histolytica/Entamoeba dispar was isolated from 50 asymptomatic amebic cyst passers in three institutions for the mentally retarded in Kanagawa Prefecture, Japan. To distinguish between E. histolytica and E. dispar, the isolates were analyzed by PCR, reactivity to monoclonal antibodies, and zymodemes. All isolates were identified as E. histolytica. The results lead us to conceive that, in Japan, E. histolytica is predominant even in asymptomatic cyst passers.

Malaria in Morong, Bataan, The Philippines, a municipality with relatively low level, but stable malaria is associated with small foci of relatively high endemicity. Although there is little association between age and symptomatic malaria, there is a reservoir of asymptomatic cases which are present throughout the year. Risk analysis suggests that the greatest risk factor in acquiring malaria depends on place of residence and not on occupation, including those associated with forest activities such as charcoal making. Foci of infection and the timing of symptomatic cases is closely correlated with breeding sites and abundance of adult Anopheles flavirostris. In spite of this close association, widely held views in the community that malaria is not related to mosquito transmission are likely to make better malaria control based on vector control difficult to sustain. Observation of treatment practices in the community and estimates of the number of apparently asymptomatic carriers from active case detection illustrate the importance of delayed treatment in providing a continuing reservoir of infection. These results highlight the need for improved early case detection and treatment.

The results of a survey on the prevalence of patent lungworm infections in herds of dairy cows in the Netherlands are presented. Low patent infections were recorded in February-March on six out of 40 farms in at least one out of 40 cows. Between mid-April and mid-June low patent infections were detected on 28 out of 39 of these farms in one to four of 40 cows. Two farms on which cows were positive in the first round were negative in the second round. One to three positive cows were found on six out of a total of 15 farms revisited in July-August. These results show that lungworm infections are cycled within herds of dairy cows in the Netherlands at a low level. This indicates that dairy cows are important as carriers for lungworm, particularly in spring. The increased patency of lungworm in cows from winter to spring may be explained by maturation of inhibited larvae.

The ability of the Babesia equi repetitive probes, pSE2 and pSB20, to detect parasites in blood from experimentally infected, naturally infected and carrier animals was tested using a spot hybridization assay. The clinical course of the experimentally infected horses was monitored using microscopy, indirect fluorescent antibody tests, packed cell volume, temperature and the probe assay. The probes sensitively monitored the parasite level during the development of the disease and correlated well with the other parameters tested. The sensitivity of the probe assay was superior to that of light microscopy, and a parasitaemia equivalent to less than 0.0025% could be detected. Detection of B. equi DNA was possible in all natural cases tested and 20 of the 119 randomly selected horses were identified as carriers of B. equi parasites. Microscopy could identify parasites in only 8 of these carrier animals. These results show that the probes can detect B. equi parasites in carrier animals and that they are suitable for use in a laboratory-based assay for B. equi.