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1.
Vet Sci ; 11(7)2024 Jul 17.
Article in English | MEDLINE | ID: mdl-39058005

ABSTRACT

Three genera of viruses of the family Totiviridae establish endosymbiotic associations with flagellated protozoa responsible for parasitic diseases of great impact in the context of One Health. Giardiavirus, Trichomonasvirus, and Leishmaniavirus infect the protozoa Giardia sp., Trichomonas vaginalis, and Leishmania sp., respectively. In the present work, we review the characteristics of the endosymbiotic relationships established, the advantages, and the consequences caused in mammalian hosts. Among the common characteristics of these double-stranded RNA viruses are that they do not integrate into the host genome, do not follow a lytic cycle, and do not cause cytopathic effects. However, in cases of endosymbiosis between Leishmaniavirus and Leishmania species from the Americas, and between Trichomonasvirus and Trichomonas vaginalis, it seems that it can alter their virulence (degree of pathogenicity). In a mammalian host, due to TLR3 activation of immune cells upon the recognition of viral RNA, uncontrolled inflammatory signaling responses are triggered, increasing pathological damage and the risk of failure of conventional standard treatment. Endosymbiosis with Giardiavirus can cause the loss of intestinal adherence of the protozoan, resulting in a benign disease. The current knowledge about viruses infecting flagellated protozoans is still fragmentary, and more research is required to unravel the intricacies of this three-way relationship. We need to develop early and effective diagnostic methods for further development in the field of translational medicine. Taking advantage of promising biotechnological advances, the aim is to develop ad hoc therapeutic strategies that focus not only on the disease-causing protozoan but also on the virus.

2.
Microorganisms ; 9(12)2021 Dec 02.
Article in English | MEDLINE | ID: mdl-34946102

ABSTRACT

Leishmaniasis is a vector-borne parasitic disease caused by Leishmania species. The disease affects humans and animals, particularly dogs, provoking cutaneous, mucocutaneous, or visceral processes depending on the Leishmania sp. and the host immune response. No vaccine for humans is available, and the control relies mainly on chemotherapy. However, currently used drugs are old, some are toxic, and the safer presentations are largely unaffordable by the most severely affected human populations. Moreover, its efficacy has shortcomings, and it has been challenged by the growing reports of resistance and therapeutic failure. This manuscript presents an overview of the currently used drugs, the prevailing model to develop new antileishmanial drugs and its low efficiency, and the impact of deconstruction of the drug pipeline on the high failure rate of potential drugs. To improve the predictive value of preclinical research in the chemotherapy of leishmaniasis, several proposals are presented to circumvent critical hurdles-namely, lack of common goals of collaborative research, particularly in public-private partnership; fragmented efforts; use of inadequate surrogate models, especially for in vivo trials; shortcomings of target product profile (TPP) guides.

3.
PLoS One ; 13(3): e0193118, 2018.
Article in English | MEDLINE | ID: mdl-29513692

ABSTRACT

Haemonchus contortus is the most pathogenic gastrointestinal helminth of small ruminants. Natural or experimental repeated infections and several native antigens confer a partially protective immune response but vaccination with subunit antigens has been elusive. Promising results have been obtained with a recombinant form of a somatic antigen (rHc23). In this paper we present the results obtained in vaccination trials in lambs using two dosages of rHc23 and standard adjuvants. Six-months old Manchego females lambs were vaccinated with rHc23 (50 or 200 µg/dose) adjuvanted with 1mL aluminum hydroxide on days -42, -28 and -14 or with 200 µg/dose rHc23 and 5 mg Quil A on days -49, -28 and -7. Control lambs were kept receiving only the adjuvants the same days or no treatment. Moreover one group did not receive any treatment or infection. On day 0 vaccinated lambs, untreated animals and those receiving the adjuvant were infected per os with a monospecific single infection of 4000 L3 of H. contortus. Infection was kept for 58 days and follow-up included the determination of serum specific antibody response (ELISA, WB), hematological parameters (eosinophil counts, hematocrit) and fecal egg counts (epg). Absence of hematocrit alterations, reduction of helminth's eggs output and abomasal parasite burden at the end of the experiment were the efficacy criteria of vaccination with the recombinant. Immunization with both adjuvants and antigen dosages elicited strong antibody responses particularly with Quil A. Vaccinated groups showed significant reduction of fecal egg excretion and abomasal helminth burdens. Highest protection of lambs against challenge was achieved with aluminum hydroxide and 200 µg/dose rHc23 with a reduction of over 70% of the abomasal burden and over 80% of fecal egg output. Results suggest that rHc23 could be a valuable recombinant candidate for vaccination against haemonchosis. No clear relationship was found between antibody levels and protection this pointing towards involvement of both humoral and cellular components in the protective response elicited by rHc23.


Subject(s)
Antigens, Helminth/immunology , Haemonchus/immunology , Nematode Infections/immunology , Sheep Diseases/immunology , Sheep/immunology , Vaccination/methods , Abomasum/immunology , Abomasum/parasitology , Adjuvants, Immunologic/administration & dosage , Animals , Antibodies, Helminth/blood , Antibodies, Helminth/immunology , Antigens, Helminth/genetics , Feces/parasitology , Female , Haemonchus/genetics , Nematode Infections/parasitology , Nematode Infections/veterinary , Parasite Egg Count , Recombinant Proteins/immunology , Sheep/parasitology , Sheep Diseases/parasitology , Sheep Diseases/prevention & control , Time Factors
4.
PLoS Negl Trop Dis ; 10(3): e0004525, 2016 Mar.
Article in English | MEDLINE | ID: mdl-27023069

ABSTRACT

BACKGROUND: Allicin has shown antileishmanial activity in vitro and in vivo. However the mechanism of action underlying its antiproliferative effect against Leishmania has been virtually unexplored. In this paper, we present the results obtained in L.infantum and a mechanistic basis is proposed. METHODOLOGY/PRINCIPAL FINDING: Exposure of the parasites to allicin led to high Ca2+ levels and mitochondrial reactive oxygen species (ROS), collapse of the mitochondrial membrane potential, reduced production of ATP and elevation of cytosolic ROS. The incubation of the promastigotes with SYTOX Green revealed that decrease of ATP was not associated with plasma membrane permeabilization. Annexin V and propidium iodide (PI) staining indicated that allicin did not induce phospholipids exposure on the plasma membrane. Moreover, DNA agarose gel electrophoresis and TUNEL analysis demonstrated that allicin did not provoke DNA fragmentation. Analysis of the cell cycle with PI staining showed that allicin induced cell cycle arrest in the G2/M phase. CONCLUSIONS/SIGNIFICANCE: We conclude that allicin induces dysregulation of calcium homeostasis and oxidative stress, uncontrolled by the antioxidant defense of the cell, which leads to mitochondrial dysfunction and a bioenergetic catastrophe leading to cell necrosis and cell cycle arrest in the premitotic phase.


Subject(s)
Calcium/metabolism , Leishmania/drug effects , Mitochondria/drug effects , Sulfinic Acids/pharmacology , Adenosine Triphosphate/metabolism , Apoptosis/drug effects , Disulfides , Dose-Response Relationship, Drug , Membrane Potential, Mitochondrial , Reactive Oxygen Species , Sulfinic Acids/administration & dosage
5.
J Antimicrob Chemother ; 69(12): 3268-74, 2014 Dec.
Article in English | MEDLINE | ID: mdl-25096077

ABSTRACT

OBJECTIVES: To evaluate the efficacy of the combination of allicin and amphotericin deoxycholate (AmB) in the chemotherapy of Leishmania infantum infection with the final aim of reducing the dose of AmB in the chemotherapy of visceral leishmaniasis. METHODS: Hamsters were intraperitoneally (ip) infected with L. infantum (10(7) stationary phase promastigotes). On day 45 post-infection animals were treated ip with AmB (1 or 5 mg/kg/day), allicin (5 mg/kg/day) or a combination of AmB (1 mg/kg/day) + allicin (5 mg/kg/day) for 5 days. Animals were clinically and biopathologically monitored and the antibody response (IgG, IgG1, IgG2) was determined. Parasite burdens were estimated by limiting dilution and AmB biodistribution was determined by HPLC in plasma, kidney, spleen and liver. RESULTS: No clinical signs or liver and kidney alterations were observed. AmB (1 mg/kg/day) did not clear the Leishmania infection and no parasites were detected in two animals treated with 5 mg/kg/day allicin. Combination therapy (5 mg/kg allicin + 1 mg/kg AmB) reduced the L. infantum burden by >95%. Antileishmanial activity of the combination was comparable (P < 0.05) to the standard AmB treatment (5 mg/kg). CONCLUSIONS: Allicin alone (5 mg/kg/day for 5 days) significantly reduced the Leishmania burden in spleen and liver of infected hamsters. Co-administration of allicin (5 mg/kg/day for 5 days) and AmB (1 mg/kg/day for 5 days) showed a partial additive effect on the reduction of leishmanial burden in both target organs.


Subject(s)
Amphotericin B/therapeutic use , Antiprotozoal Agents/therapeutic use , Leishmania infantum/drug effects , Leishmaniasis, Visceral/drug therapy , Sulfinic Acids/therapeutic use , Amphotericin B/pharmacokinetics , Animal Structures/chemistry , Animals , Antiprotozoal Agents/pharmacokinetics , Chromatography, High Pressure Liquid , Cricetinae , Disease Models, Animal , Disulfides , Drug Therapy, Combination/methods , Female , Leishmania infantum/isolation & purification , Parasite Load , Plasma/chemistry , Sulfinic Acids/pharmacokinetics , Treatment Outcome
6.
Int J Parasitol ; 44(7): 429-36, 2014 Jun.
Article in English | MEDLINE | ID: mdl-24759430

ABSTRACT

A somatic protein from adult Haemonchus contortus (Hc23), the most abundant component in a low molecular weight fraction with known immunizing effect against experimental haemonchosis, has been purified by immunochromatography. The immunoprophylactic value of Hc23 was tested in groups of 5-6 months old Assaf lambs using Al(OH)(3) or Escherichia coli lipopolysaccharide+inactivated Propionibacterium acnes as adjuvant and the results compared with uninfected control, uninfected and challenged or infected and challenged lambs. Immunization with Hc23 in either adjuvant elicited significant reductions in fecal egg counts after challenge with 15,000 L3s (70.67%-85.64%, respectively) and reduced (67.1% and 86%) abomasal worm counts (45 days post-challenge). Immunized lambs displayed higher peripheral eosinophil counts, were less anaemic and had weight gains than challenged controls. The results suggest that the Hc23 antigen can induce a partially protective response against haemonchosis in lambs.


Subject(s)
Haemonchiasis/veterinary , Haemonchus/immunology , Helminth Proteins/immunology , Sheep Diseases/prevention & control , Vaccines/immunology , Amino Acid Sequence , Animals , Antigens, Helminth/chemistry , Antigens, Helminth/genetics , Antigens, Helminth/immunology , Feces/parasitology , Haemonchiasis/parasitology , Haemonchiasis/prevention & control , Helminth Proteins/chemistry , Helminth Proteins/genetics , Parasite Egg Count/veterinary , Sheep , Sheep Diseases/parasitology
7.
Vet Parasitol ; 203(1-2): 231-6, 2014 Jun 16.
Article in English | MEDLINE | ID: mdl-24636785

ABSTRACT

Intestinal helminth infections are common and of paramount economic importance in domestic animals. Available chemotherapy is limited and anthelmintic resistance is widespread in some hosts. This scenario favors the exploration of alternative methods of control, among them immune modulators. The effect of Escherichia coli LPS+Propionibacterium acnes on a primary infection of Heligmosomoides polygyrus (Trichostongyloidea) in mice has been tested. Nematode infection induced a rise of specific IgG1, both serum and intestinal, and a significant reduction in the unspecific (ConA) lymphoproliferative response. Treatment with the immune modulator (days -2, 0, 7 and 14 post infection) elicited an apparent delay of larval intramucosal development. Moreover cumulative nematode egg shedding in treated mice was significantly lower (p=0.0041). Preliminary results point toward the interest of immune modulators to control intestinal helminths.


Subject(s)
Antibody Formation/immunology , Lipopolysaccharides/pharmacology , Propionibacterium acnes/immunology , Strongylida Infections/immunology , Animals , Antibodies, Helminth/analysis , Antibodies, Helminth/blood , Antibody Formation/drug effects , Feces/parasitology , Heligmosomatoidea/immunology , Immunologic Factors/pharmacology , Immunomodulation/immunology , Intestines/parasitology , Intestines/pathology , Larva , Mice, Inbred BALB C , Parasite Egg Count , Parasite Load
8.
J Parasitol ; 94(5): 1031-7, 2008 Oct.
Article in English | MEDLINE | ID: mdl-18576697

ABSTRACT

A study on age distribution and seasonal dynamics of abomasal helminths in wild red deer was conducted in central Spain, by monthly samplings of fawns (<1 yr), subadult (1-2 yr), and adult (>2 yr) animals. Both intensity and prevalence of abomasal parasitism were higher in older animals, particularly in males. A bimodal pattern for intensity of infection by gastrointestinal parasites was observed. Maximum values attained in winter and summer may be related to variation in climate and the shifting availability of forage resources. The pattern was largely due to the contribution of Spiculopteragia asymmetrica/Spiculopteragia quadrispiculata, whereas the other species found (Ostertagia leptospicularis/Ostertagia kolchida and Ostertagia drozdzi/Ostertagia ryjikovi) occurred with lower prevalence and intensity of infection. Among these ostertagiines, the ratio for major and minor morphotypes of males of respective species and the relative abundance of males and females were stable through the annual cycle.


Subject(s)
Abomasum/parasitology , Deer/parasitology , Helminthiasis, Animal/epidemiology , Stomach Diseases/veterinary , Age Distribution , Animals , Animals, Wild , Female , Helminthiasis, Animal/parasitology , Male , Ostertagiasis/epidemiology , Ostertagiasis/parasitology , Ostertagiasis/veterinary , Prevalence , Seasons , Sex Distribution , Spain/epidemiology , Stomach Diseases/epidemiology , Stomach Diseases/parasitology , Trichostrongyloidiasis/epidemiology , Trichostrongyloidiasis/parasitology , Trichostrongyloidiasis/veterinary
9.
Rev. cient. (Maracaibo) ; 17(6): 577-587, nov.-dic. 2007.
Article in English | LILACS | ID: lil-548564

ABSTRACT

La hemoncosis, causada por el nematodo del abomaso de rumiantes Haemonchus contortus, constituye una de las enfermedades parasitarias más notables del ganado ovino en todo el mundo. Las infecciones provocan síndromes anémicos y de mala digestión/absorción que pueden causar la muerte en los casos agudos y disminución de la producción en las formas crónicas. Las claves principales para la aparición de esta enfermedad son el comportamiento biológico del helmito, su patogenia, además de la respuesta del hospedador. El conocimiento actualizado de estos aspectos permitirá una mayor eficaciencia de los métodos de diagnóstico y contro del proceso y, como consecuencia, la disminución de los riesgos de aparición de esta enfermedad. La presente revisión analiza y discute algunos aspectos de la relación ganado ovino-H. contortus de interés para disminuir el impacto de esta parasitosis sobre la salud y producción ganaderas.


Haemonchosis, caused by the abomasal nematode Haemonchus contortus, is among the most relevant parasitic diseases of small ruminants all over the world. The infections are responsible for anemic and bad digestion/absorption syndromes causing the death of severely infected animals in acute cases and the reduction of production scores in the chronic forms of the disease. The keys for the appearance of the disease include the biological behavior of the helminth and mechanisms of pathogenesis, besides the host’s response. The updated knowledge of these aspects would result in a higher efficiency of diagnostic and control methods, thus reducing the risks of appearance of the disease. In the present review, several aspects of the sheep - H. contortus relationship are analyzed with the aim of reducing the impact of this parasitosis on livestock health and productivity.


Subject(s)
Animals , Digestion , Parasitic Diseases, Animal/pathology , Haemonchus/pathogenicity , Haemonchiasis/parasitology , Haemonchiasis/veterinary , Sheep/parasitology , Parasitology , Veterinary Medicine
10.
FEMS Microbiol Lett ; 267(1): 9-16, 2007 Feb.
Article in English | MEDLINE | ID: mdl-17233673

ABSTRACT

We have characterized diverse strains or species of Leishmania isolated in humans that are currently circulating throughout Peru, by means of isoenzymatic characterization, kDNA analysis by restriction enzymes, and multiplex PCR assay. The cluster analysis gave five groups. Cluster 1 includes L. (L.) donovani together with the isolates LP4 and LP7, forming the donovani complex. Thus, this complex corresponds to the New World visceral form, L. (L.) chagasi. Cluster 2 is formed by the isolates LP1-LP3, LP6, LP10, LP9, and LP11, phylogenetically intermediate between Cluster 1 and Cluster 3, or they can be treated as hybrids. Cluster 3 is divided into two subgroups: one formed by L. (V.) peruviana, together with the isolates LP14 and LP5, and the second one formed by L. (V.) brazilensis and the isolate LP8. These two subgroups form part of the brazilensis complex. The three strains of L. (L.) infantum [L. (L.) infantum I and II and la LSI] make up Cluster 4. In Cluster 5, we include the three Mexican strains (LM1-LM3) forming one subgroup while we would place L. (L.) amazonensis in another subgroup. These two subgroups would comprise the complex mexicana.


Subject(s)
DNA, Kinetoplast/metabolism , Leishmania/classification , Leishmania/isolation & purification , Leishmaniasis/parasitology , Polymerase Chain Reaction/methods , Animals , Cluster Analysis , DNA Fingerprinting , DNA Restriction Enzymes/metabolism , DNA, Kinetoplast/genetics , Isoelectric Focusing , Isoenzymes , Leishmania/genetics , Peru , Polymorphism, Restriction Fragment Length , Protozoan Proteins/analysis , Protozoan Proteins/isolation & purification
11.
J Parasitol ; 90(6): 1378-86, 2004 Dec.
Article in English | MEDLINE | ID: mdl-15715232

ABSTRACT

A survey of abomasal parasites in cervids from Central Spain was conducted at 3 sites, Quintos de Mora (Toledo), Maluéñez de Arriba (Cáceres), and La Herguijuela (Cáceres). Commonly occurring helminths belonged to 3 polymorphic species of the Ostertagiinae: Spiculopteragia asymmetricalS. quadrispiculata, Ostertagia leptospicularis/O. kolchida, and O. drozdzi/O. ryjikovi. Trichostrongylus axei was found in very few cases. Ostertagia drozdzi/O. ryjikovi and the minor male morphotype, S. quadrispiculata, are reported for the first time in red deer from Spain. The 3 ostertagiine species are also reported for the first time in fallow deer from Spain. These 3 species of Ostertagiinae are primarily parasites of cervids, and nematode species characteristic of domestic ruminants were not present. Prevalence of infection by gastrointestinal parasites in cervids was high, ranging from 97.5 to 100%, across the 3 areas sampled. Mean intensity of infection and abundance showed a positive relationship to the population density of red deer. Helminth burdens were higher in fallow deer than in the sympatric red deer and may reflect the gregarious social structure and different foraging patterns of fallow deer.


Subject(s)
Abomasum/parasitology , Deer/parasitology , Nematode Infections/veterinary , Stomach Diseases/veterinary , Animal Husbandry/methods , Animals , Female , Male , Nematode Infections/epidemiology , Nematode Infections/parasitology , Prevalence , Sex Distribution , Spain/epidemiology , Stomach Diseases/epidemiology , Stomach Diseases/parasitology
12.
Acta Trop ; 84(3): 175-81, 2002 Dec.
Article in English | MEDLINE | ID: mdl-12443795

ABSTRACT

Fourteen dogs naturally infected with Leishmania infantum and treated with allopurinol were monitored clinically and serologically with immunofluorescent antibody test (IFAT, amastigotes and promastigotes), enzyme linked-immunosorbent-assay (ELISA, IgG1 and IgG2) and Western blotting (WB). In all dogs therapy lead to clinical improvement together with decreasing specific antibodies in IFAT, ELISA and WB, demonstrating the usefulness of serology for follow-up. Although IgG1 and IgG2 varied considerably between individual animals, IgG2 of all dogs was predominantly in both ELISA and WB. This suggests the value of monitoring the IgG2 response (especially against 29 and 67 kDa antigens) in the follow-up of treated dogs.


Subject(s)
Allopurinol/therapeutic use , Antiprotozoal Agents/therapeutic use , Dog Diseases/parasitology , Leishmania infantum/pathogenicity , Leishmaniasis, Visceral/veterinary , Animals , Blotting, Western/methods , Dog Diseases/diagnosis , Dog Diseases/drug therapy , Dogs , Enzyme-Linked Immunosorbent Assay/methods , Fluorescent Antibody Technique, Direct/methods , Follow-Up Studies , Host-Parasite Interactions , Immunoglobulin G/analysis , Leishmania infantum/cytology , Leishmaniasis, Visceral/diagnosis , Leishmaniasis, Visceral/drug therapy , Treatment Outcome
13.
Clin Diagn Lab Immunol ; 9(5): 1137-41, 2002 Sep.
Article in English | MEDLINE | ID: mdl-12204974

ABSTRACT

This study of several techniques for detecting cryptic leishmaniasis in dogs from areas in Spain where Leishmania infantum is highly endemic concludes that immunological techniques (enzyme-linked immunosorbent assay, immunofluorescence antibody test, Western blotting, delayed-type hypersensitivity reaction, and in vitro lymphocyte proliferation assay) do not clearly differentiate between noninfected and infected asymptomatic dogs and that culture and PCR are more reliable diagnostic tools.


Subject(s)
Dog Diseases/diagnosis , Enzyme-Linked Immunosorbent Assay , Leishmania infantum/isolation & purification , Leishmaniasis/diagnosis , Animals , Blotting, Western , Carrier State , Dog Diseases/immunology , Dog Diseases/parasitology , Dogs , Fluorescent Antibody Technique , Hypersensitivity, Delayed/diagnosis , Hypersensitivity, Delayed/parasitology , Leishmaniasis/immunology
14.
J Parasitol ; 88(2): 417-8, 2002 Apr.
Article in English | MEDLINE | ID: mdl-12054028

ABSTRACT

Sequences of ITS-1 and ITS-2 rDNA for adult males of Spiculopteragia asymmetrica and Spiculopteragia quadrispiculata in red deer (Cervus elaphus) were determined. They were found to be identical, suggesting that S. asymmetrica and S. quadrispiculata represent a single species and do not refute the concept of dimorphic species in the Spiculopteragia.


Subject(s)
DNA, Helminth/genetics , DNA, Ribosomal Spacer/genetics , Deer/parasitology , Trichostrongyloidea/genetics , Animals , Base Sequence , DNA, Helminth/chemistry , DNA, Ribosomal Spacer/chemistry , Male , Molecular Sequence Data , Polymerase Chain Reaction , Sequence Homology, Nucleic Acid , Species Specificity , Trichostrongyloidea/chemistry , Trichostrongyloidea/classification
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