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Ancylostoma duodenale [keywords]
- Evaluation of Biochemical, Hematological and Parasitological Parameters of Protein-Deficient Hamsters Infected with Ancylostoma ceylanicum. [Journal Article]
- PLoS Negl Trop Dis 2014 Sep; 8(9):e3184.
Hookworms infect millions of people worldwide and can cause severe clinical symptoms in their hosts. Prospective cohort studies in Brazil show high rates of hookworm reinfection in malnourished children compared to well-nourished children, despite previous treatment. Additionally, soil-transmitted helminth (STH) infections can worsen the nutritional status of affected populations. Therefore, this study aims to clarify the effects of host malnutrition during Ancylostoma ceylanicum infection and how this infection affects host physiological parameters using a hamster model.Hamsters were divided into four experimental groups: normal diet or low-protein diet (also referred to as "malnourished") and A. ceylanicum infection or no infection. More severe pathogenesis was observed in the infected malnourished group, as demonstrated by significant decreases in the hemoglobin concentration, erythrocyte number and packed-cell volume compared to the non-infected malnourished group. Greater numbers of adult parasites and eggs were observed in the malnourished group compared to the control group; however, the oviposition rate was lower in the malnourished group. In general, greater values of total lipids were observed in malnourished animals compared to control animals, including lipids excreted in the stool.In this work, we have demonstrated that animals fed an isocaloric low-protein diet presented more severe pathogenesis when infected with A. ceylanicum. The increased lipid concentration in the liver and blood is related to the conversion of the excess carbohydrate into fatty acids that increase the concentration of triglycerides in general. Triglycerides were excreted in the feces, indicating that infection associated with malnutrition caused a greater loss of these molecules for this group of animals and confirming the hypothesis that both nutrition and infection are responsible for the malabsorption syndrome. Taken together, the results found in this work confirm the hypothesis that the nutritional condition of the host greatly influences the course of the infection.
- Gastrointestinal parasites in rural dogs and cats in Selangor and Pahang states in Peninsular Malaysia. [Journal Article]
- Acta Parasitol 2014 Oct; 59(4):737-44.
To estimate the current prevalence of gastrointestinal (GI) parasites in dogs and cats, a total of 105 fresh faecal samples were collected from rural areas in Peninsular Malaysia. Each faecal sample was examined for the presence of GI parasites by microscopic examination after formalin-ether concentration technique and for protozoa, trichrome and Ziehl-Neelsen staining were employed. The overall prevalence of GI parasitic infection was 88.6% (95% CI = 82.5-94.7) in which 88.3% of dogs and 89.3% of cats were infected with at least one parasites species, respectively. There were 14 different GI parasites species (nematodes, cestodes and protozoa) detected, including Ancylostoma spp. (62.9%), Toxocara spp. (32.4%), Trichuris vulpis (21.0%), Spirometra spp. (9.5%), Toxascaris leonina (5.7%), Dipylidium caninum (4.8%), Ascaris spp. (2.9%), Hymenolepis diminuta (1.0%) and others. General prevalence of GI parasites showed a significant difference between helminth (84.4%) and protozoa (34.3%) infections. Monoparasitism (38.1%) was less frequent than polyparasitism (46.7%). As several of these GI parasites are recognized as zoonotic agents, the results of this investigation revealed that local populations may be exposed to a broad spectrum of zoonotic agents by means of environmental contamination with dogs and cats faeces and this information should be used to mitigate public health risks. Prevention and control measures have to be taken in order to reduce the prevalence rates especially in socioeconomically disadvantaged communities where animals live in close proximity to people, poor levels of hygiene and overcrowding together with a lack in veterinary attention and zoonotic awareness.
- Beta carbonic anhydrases: novel targets for pesticides and anti-parasitic agents in agriculture and livestock husbandry. [JOURNAL ARTICLE]
- Parasit Vectors 2014 Aug 29; 7(1):403.
The genomes of many insect and parasite species contain beta carbonic anhydrase (beta-CA) protein coding sequences. The lack of beta-CA proteins in mammals makes them interesting target proteins for inhibition in treatment of some infectious diseases and pests. Many insects and parasites represent important pests for agriculture and cause enormous economic damage worldwide. Meanwhile, pollution of the environment by old pesticides, emergence of strains resistant to them, and their off-target effects are major challenges for agriculture and society.In this study, we analyzed a multiple sequence alignment of 31 beta-CAs from insects, some parasites, and selected plant species relevant to agriculture and livestock husbandry. Using bioinformatics tools a phylogenetic tree was generated and the subcellular localizations and antigenic sites of each protein were predicted. Structural models for beta-CAs of Ancylostoma caninum, Ascaris suum, Trichinella spiralis, and Entamoeba histolytica, were built using Pisum sativum and Mycobacterium tuberculosis beta-CAs as templates.Six beta-CAs of insects and parasites and six beta-CAs of plants are predicted to be mitochondrial and chloroplastic, respectively, and thus may be involved in important metabolic functions. All 31 sequences showed the presence of the highly conserved beta-CA active site sequence motifs, CXDXR and HXXC (C: cysteine, D: aspartic acid, R: arginine, H: histidine, X: any residue). We discovered that these two motifs are more antigenic than others. Homology models suggested that these motifs are mostly buried and thus not well accessible for recognition by antibodies.The predicted mitochondrial localization of several beta-CAs and hidden antigenic epitopes within the protein molecule, suggest that they may not be considered major targets for vaccines. Instead, they are promising candidate enzymes for small-molecule inhibitors which can easily penetrate the cell membrane. Based on current knowledge, we conclude that beta-CAs are potential targets for development of small molecule pesticides or anti-parasitic agents with minimal side effects on vertebrates.
- [Cutaneous larva migrans after a trip to the Caribean]. [English Abstract, Journal Article]
- Rev Chilena Infectol 2014 Jun; 31(3):346-8.
Cutaneous larva migrans is a parasitic disease caused by Ancylostoma braziliense and Ancylostoma caninum larvae, which is transmitted by contact with sand infested with these parasites. Dogs and cats are the definitive hosts. This parasitic disease is endemic in the Caribbean, Africa, Australia, and Asia. We present the case of a 27-year-old woman, who developed skin lesions compatible with cutaneous larva migrans on her right foot after returning from beach vacations in the Mexican Caribbean. After clinical diagnosis, oral ivermectin was administered, with good clinical response.
- The prevalence and distribution of gastrointestinal parasites of stray and refuge dogs in four locations in India. [JOURNAL ARTICLE]
- Vet Parasitol 2014 Jul 30.
A gastrointestinal parasite survey of 411 stray and refuge dogs sampled from four geographical and climactically distinct locations in India revealed these animals to represent a significant source of environmental contamination for parasites that pose a zoonotic risk to the public. Hookworms were the most commonly identified parasite in dogs in Sikkim (71.3%), Mumbai (48.8%) and Delhi (39.1%). In Ladakh, which experiences harsh extremes in climate, a competitive advantage was observed for parasites such as Sarcocystis spp. (44.2%), Taenia hydatigena (30.3%) and Echinococcus granulosus (2.3%) that utilise intermediate hosts for the completion of their life cycle. PCR identified Ancylostoma ceylanicum and Ancylostoma caninum to occur sympatrically, either as single or mixed infections in Sikkim (Northeast) and Mumbai (West). In Delhi, A. caninum was the only species identified in dogs, probably owing to its ability to evade unfavourable climatic conditions by undergoing arrested development in host tissue. The expansion of the known distribution of A. ceylanicum to the west, as far as Mumbai, justifies the renewed interest in this emerging zoonosis and advocates for its surveillance in future human parasite surveys. Of interest was the absence of Trichuris vulpis in dogs, in support of previous canine surveys in India. This study advocates the continuation of birth control programmes in stray dogs that will undoubtedly have spill-over effects on reducing the levels of environmental contamination with parasite stages. In particular, owners of pet animals exposed to these environments must be extra vigilant in ensuring their animals are regularly dewormed and maintaining strict standards of household and personal hygiene.
- Identification of Angiostrongylus cantonensis and other nematodes using the SSU rDNA in Achatina fulica populations of Metro Manila. [Journal Article]
- Trop Biomed 2014 Jun; 31(2):327-35.
Angiostrongylus cantonensis is a parasitic nematode that causes eosinophilic meningitis in humans. Accidental infection occurs by consumption of contaminated intermediates, such as the giant African land snail, Achatina fulica. This study surveyed the presence of A. cantonensis juveniles in A. fulica populations from 12 sites in Metropolitan Manila, Philippines using the SSU rDNA. Fourteen distinct sequences from 226 nematodes were obtained; of these, two matched A. cantonensis and Ancylostoma caninum, respectively, with 100% identity. Exact identities of the remaining twelve sequences could not be determined due to low percent similarities. Of the sequenced nematodes, A. cantonensis occurred with the highest frequency (139 out of 226). Most of these (131 out of 139) were collected in just one area in Quezon City. Nematode infection of A. fulica in this area and two others from Makati and another area in Quezon City, respectively, were highest, combining for 95% of the total infection. Ancylostoma caninum, on the other hand, was detected in four different sites. A. caninum is a canine parasite, and this is the first report of the nematode in A. fulica. These results cause public health concerns as both A. cantonensis and A. caninum are zoonotic to humans.
- Efficacy of milbemycin oxime in combination with spinosad in the treatment of larval and immature adult stages of Ancylostoma caninum and Toxocara canis in experimentally infected dogs. [JOURNAL ARTICLE]
- Vet Parasitol 2014 Jul 28.
Ancylostoma caninum and Toxocara canis are two important zoonotic parasites of dogs. The primary objective of these studies were to confirm the oral effectiveness of milbemycin oxime (MO) and spinosad in dogs experimentally infected with immature (L4 and immature adult) stages of T. canis or A. caninum. Both trials were conducted as randomized, blinded, placebo-controlled dose confirmation studies. Treatments using the intended European commercial tablet formulation of Trifexis were administered in a timeframe relative to inoculation so that effectiveness could be assessed against specific immature stages of A. caninum or T. canis. In each study on Day 0, each of 32, 3-4 month old dogs were inoculated with 250 infective eggs of T. canis or 300 infective L3 of the hookworm, A. caninum. All dogs were weighed before their scheduled treatment, randomized to 1 of the 4 treatment groups in each study (8 dogs/group). All dogs were fed just prior to dosing. For T. canis, dogs were treated orally with an MO/spinosad tablet on Day 14 or Day 24. For A. caninum, dogs were treated orally with an MO/spinosad tablet on Day 7 or Day 11. Corresponding control groups in each study received a placebo tablet. Dogs were necropsied 5 or 6 days after their respective treatments. The digestive tract was removed and processed to recover, count, and identify all stages. The GM worm count for the MO/spinosad tablet on Day 14 (L4 T. canis) was 0.0, with efficacy calculated as 100%; however, only 3 of 8 control dogs had adequate infections. The GM worm count for the MO/spinosad tablet on Day 24 (immature adult stage) was 0.30; efficacy calculated at 96.15%. This is based on 5 of the 8 control dogs with adequate infections. In the two A. caninum studies, GM worm counts for the MO/spinosad tablets on Day 7 (L4 efficacy) was 2.37 and 0.8 with efficacy calculated as 98.92% and 99.25%, respectively. The GM count for the group treated with the MO/spinosad combination on Day 11 (immature adult) was 6.19 and 1.4; efficacy calculated at 97.77% and 98.58%, respectively. A minimum MO oral dose of 0.75mg/kg was highly effective for the treatment of immature stages of T. canis and A. caninum infections in dogs. The ability to kill immature stages of these two parasites before they become patent will benefit dogs, their owners and family members due to reduced exposure to these potentially zoonotic parasites.
- Molecular cloning and analysis of Ancylostoma ceylanicum glutamate-cysteine ligase. [JOURNAL ARTICLE]
- Mol Biochem Parasitol 2014 Aug 1.
Glutamate-cysteine ligase (GCL) is a heterodimer enzyme composed of a catalytic subunit (GCLC) and a modifier subunit (GCLM). This enzyme catalyses the synthesis of γ-glutamylcysteine, a precursor of glutathione. cDNAs of the putative glutamate-cysteine ligase catalytic (Ace-GCLC) and modifier subunits (Ace-GCLM) of Ancylostoma ceylanicum were cloned using the RACE-PCR amplification method. The Ace-gclc and Ace-gclm cDNAs encode proteins with 655 and 254 amino acids and calculated molecular masses of 74.76 and 28.51kDa, respectively. The Ace-GCLC amino acid sequence shares about 70% identity and 80% sequence similarity with orthologs in Loa loa, Onchocerca volvulus, Brugia malayi, and Ascaris suum, whereas the Ace-GCLM amino acid sequence has only about 30% sequence identity and 50% similarity to homologous proteins in those species. Real-time PCR analysis of mRNA expression in L3, serum stimulated L3 and adult stages of A. ceylanicum showed the highest level of Ace-GCLC and Ace-GCLM expression occurred in adult worms. No differences were detected among adult hookworms harvested 21 and 35dpi indicating expression of Ace-gclc and Ace-gclm in adult worms is constant during the course of infection. Positive interaction between two subunits of glutamate-cysteine ligase was detected using the yeast two-hybrid system, and by specific enzymatic reaction. Ace-GCL is an intracellular enzyme and is not exposed to the host immune system. Thus, as expected, we did not detect IgG antibodies against Ace-GCLC or Ace-GCLM on days 21, 60 and 120 of A. ceylanicum infection in hamsters. Furthermore, vaccination with one or both antigens did not reduce worm burdens, and resulted in no improvement of clinical parameters (hematocrit and hemoglobin) of infected hamsters. Therefore, due to the significant role of the enzyme in parasite metabolism, our analyses raises hope for the development of a successful new drug against ancylostomiasis based on the specific GCL inhibitor.
- Schistosoma mansoni venom allergen-like protein 4 (SmVAL4) is a novel lipid-binding SCP/TAPS protein that lacks the prototypical CAP motifs. [JOURNAL ARTICLE]
- Acta Crystallogr D Biol Crystallogr 2014 Aug 1; 70(Pt 8):2186-2196.
Schistosomiasis is a parasitic disease that affects over 200 million people. Vaccine candidates have been identified, including Schistosoma mansoni venom allergen-like proteins (SmVALs) from the SCP/TAPS (sperm-coating protein/Tpx/antigen 5/pathogenesis related-1/Sc7) superfamily. The first SmVAL structure, SmVAL4, was refined to a resolution limit of 2.16 Å. SmVAL4 has a unique structure that could not be predicted from homologous structures, with longer loops and an unusual C-terminal extension. SmVAL4 has the characteristic α/β-sandwich and central SCP/TAPS cavity. Furthermore, SmVAL4 has only one of the signature CAP cavity tetrad amino-acid residues and is missing the histidines that coordinate divalent cations such as Zn(2+) in other SCP/TAPS proteins. SmVAL4 has a cavity between α-helices 1 and 4 that was observed to bind lipids in tablysin-15, suggesting the ability to bind lipids. Subsequently, SmVAL4 was shown to bind cholesterol in vitro. Additionally, SmVAL4 was shown to complement the in vivo sterol-export phenotype of yeast mutants lacking their endogenous CAP proteins. Expression of SmVAL4 in yeast cells lacking endogenous CAP function restores the block in sterol export. These studies suggest an evolutionarily conserved lipid-binding function shared by CAP proteins such as SmVAL4 and yeast CAP proteins such as Pry1.
- Assessing the speed of kill of hookworms, Ancylostoma caninum, by Advantage Multi(®) for Dogs using endoscopic methods. [JOURNAL ARTICLE]
- Vet Parasitol 2014 May 23.
Endoscopic capsules and endoscopy were used to assess the speed of kill and the clearance of hookworms in dogs experimentally infected with Ancylostoma caninum. A total of four adult dogs were inoculated in two separate cohorts comprised of two 4-year-old females and two 7-year-old males. Dogs were treated topically with Advantage Multi(®) for Dogs 13 days (Cohort 1) or 16 days (Cohort 2) after infection. Endoscopic imaging of the small intestine was carried out both pre- and post-treatment. Examination of the first cohort revealed that the worms had been cleared and the hookworm-induced lacerations were markedly diminished within 48h of treatment. In the second cohort, endoscopic capsules were given the day of, the day after, and two days after treatment; within 24h of product administration, the worms had been removed with a concurrent reduction in observed lesions. Topical application of Advantage Multi(®) for Dogs rapidly removed worms from the small intestine of the dogs in this study as early as 24h post-treatment, with a marked reduction in the number of mucosal lesions seen.