Correlation between hematological parameters and ancylostomiasis: A retrospective study.

OBJECTIVE: Our aim intended to determine the relationship between hematological parameters (neutrophil-to-lymphocyte ratio [NLR], platelet-to-lymphocyte ratio [PLR], and eosinophil-to-lymphocyte ratio [ELR]) and ancylostomiasis. METHODS: There were 140 patients with ancylostomiasis and 159 healthy controls enrolled in this study. All data were collected from electronic medical records of the First Affiliated Hospital of Guangxi Medical University. RESULTS: The levels of NLR, PLR, and ELR in ancylostomiasis patients were significantly higher than those in the healthy controls (all P = 0.000). A receiver operating characteristic curve was generated to assess the diagnostic efficacy of these three hematological parameters. ELR (AUC = 0.850; sensitivity = 75.00%; specificity = 86.80%) showed the superior AUC than those of NLR (AUC = 0.718; sensitivity = 53.57%; specificity = 88.68%) and PLR (AUC = 0.806; sensitivity = 68.57%; specificity = 86.79%), respectively. A multivariate regression model using the two selected indices (RBC and ELR) was established with the model's sensitivity and specificity reached 82.86% and 96.23%, respectively. In the ancylostomiasis patient group, NLR (r = -0.452, P = 0.000) and PLR (r = -0.357, P = 0.000) were reversely associated with eosinophils. CONCLUSION: The pretreatment levels of the three hematological parameters (NLR, PLR, and ELR) may serve as valuable indicators for distinguishing patients with ancylostomiasis from healthy controls. NLR and PLR are negatively associated with the previous indicator, eosinophils.

Changes in biochemical analytes in female dogs with subclinical Ancylostoma spp. infection.

BACKGROUND: Ancylostoma spp. is one of the most prevalent canine intestinal nematode infections which usually causes subclinical disease in adult dogs and has zoonotic implications. Therefore, the aim of this study was to explore and evaluate the possible pathophysiological changes that Ancylostoma spp. could produce in female dogs naturally infected but without clinical signs of disease, by screening a wide variety of biochemical markers for potential changes. Samples of feces and blood of 45 dogs were collected and fecal flotation and zinc sulphate centrifugal flotation were performed. The biochemical analytes determined were: the acute-phase proteins C-reactive protein (CRP) and haptoglobin (Hp); the lipid profile (cholesterol, triglycerides, HDL, LDL); the serum iron profile: iron, unsaturated iron binding-capacity (UIBC), and ferritin; the enzyme butyrylcholinesterase (BChe); the pancreatic profile: amylase, lipase, and trypsin-like immunoreactivity (TLI); the oxidative stress markers: total antioxidant capacity (TAC) and paraoxonase -1 (PON-1), along with total protein, albumin, and insulin-like growth factor - 1 (IGF - 1). Ancylostoma spp. eggs were detected in 29/45 dogs (64.4 %). Dogs were divided into two groups according to the results of fecal flotation methods. Group 1: negative fecal floatation (n = 16), and Group 2: subclinical infection with the observation of Ancylostoma spp. type eggs/x 40 objective fields (n = 29). RESULTS: Mann-Whitney U test was used to compare the biochemical analyte results between the two groups (P < 0.05). Significant increases in CRP (µg/mL) (median): non-infected dogs: 5.5; subclinically infected dogs 18.7; P = 0.03, Hp (g/L) (median): G1: 2.4; G2: 3.3; P = 0.03, and UIBC (µg/dL) (median): non-infected dogs: 139.4; subclinically infected dogs: 216; P = 0.0015, and significantly decreased iron (µg/dL) (median): non-infected dogs: 202.5; subclinically infected dogs: 125.7; P = 0.0041, IGF-1 (ng/mL) (median): non-infected dogs: 224; subclinically infected dogs: 123; P = 0.02, and albumin (g/dL) (median): non-infected dogs: 2.8; subclinically infected dogs: 2.5; P = 0.04 concentrations were observed in dogs with subclinical Ancylostoma spp. infection when compared to non-infected dogs. CONCLUSION: These findings provide an overview of the biochemical effects produced by patent Ancylostoma spp. in naturally infected dogs without any evident clinical signs of disease, which could be considered in differential diagnosis, especially in an endemic area for this parasite.

Ivermectin treatment of free-ranging endangered Australian sea lion (Neophoca cinerea) pups: effect on hookworm and lice infection status, haematological parameters, growth, and survival.

A placebo-controlled study was used to investigate the effectiveness of ivermectin to treat hookworm (Uncinaria sanguinis) and lice (Antarctophthirus microchir) infections in free-ranging Australian sea lion (Neophoca cinerea) pups and to test the hypotheses that these parasitic infections cause anaemia, systemic inflammatory responses, and reduced growth, and contribute towards decreased pup survival. Ivermectin was identified as an effective and safe anthelmintic in this species. Pups administered ivermectin had significantly higher erythrocyte counts and significantly lower eosinophil counts compared to controls at 1-2 months post-treatment, confirming that U. sanguinis and/or A. microchir are causatively associated with disease and demonstrating the positive effect of ivermectin treatment on clinical health parameters. Higher growth rates were not seen in ivermectin-treated pups and, unexpectedly, relatively older pups treated with ivermectin demonstrated significantly reduced growth rates when compared to matched saline-control pups. Differences in survival were not identified between treatment groups; however, this was attributed to the unexpectedly low mortality rate of recruited pups, likely due to the unintended recruitment bias towards pups >1-2 months of age for which mortality due to hookworm infection is less likely. This finding highlights the logistical and practical challenges associated with treating pups of this species shortly after birth at a remote colony. This study informs the assessment of the use of anthelmintics as a tool for the conservation management of free-ranging wildlife and outlines essential steps to further the development of strategies to ensure the effective conservation of the Australian sea lion and its parasitic fauna.

Evaluation of biochemical, hematological and parasitological parameters of protein-deficient hamsters infected with Ancylostoma ceylanicum.

BACKGROUND: 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. METHODOLOGY/PRINCIPAL FINDINGS: 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. CONCLUSIONS: 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.

Ac-AP-12, a novel factor Xa anticoagulant peptide from the esophageal glands of adult Ancylostoma caninum.

Immunoscreening an Ancylostoma caninum cDNA library with canine hookworm-infected dog serum resulted in the isolation of a 461 bp cDNA encoding Ac-AP-12, a new 9.1 kDa anticoagulant peptide (100 amino acids) with 43-69% amino acid homology to other nematode anticoagulant peptides (NAPs) from Ancylostoma hookworms. Messenger RNA transcription and expression of Ac-AP-12 was unique to the adult stage of A. caninum. The yeast expressed recombinant Ac-AP-12 demonstrated potent anticoagulant activity on human blood plasma in a concentration dependent manner, and was shown to specifically inhibit human factor Xa activity. Immunolocalization with specific rabbit antiserum showed that Ac-AP-12 was exclusively located in the esophageal glands of adult hookworm. Ac-AP-12 is hypothesized to facilitate both parasite blood feeding and digestion.

An anticoagulant peptide from the human hookworm, Ancylostoma duodenale that inhibits coagulation factors Xa and XIa.

A full-length cDNA encoding an anticoagulant peptide, named AduNAP4, was cloned and identified from the human hookworm Ancylostoma duodenale. AduNAP4 has 104 amino acids including a predicted 23-residue signal peptide and shows

Role for nitric oxide in hookworm-associated immune suppression.

Hookworm infection is a major cause of anemia and malnutrition in resource-poor countries. Human and animal studies suggest that infection with these intestinal nematodes is associated with impaired cellular immunity, characterized by reduced lymphocyte proliferation in response to both parasite and heterologous antigens. We report here data from studies aimed at defining mechanisms through which hookworms modulate the host cellular immune response. Splenocytes and mesenteric lymph node (MLN) cells from hamsters infected with Ancylostoma ceylanicum showed minimal proliferation in response to mitogen at days 20 and 30 postinfection (p.i.), with partial recovery noted at day 70 p.i. The proliferative capacity of enriched splenocyte T-cell preparations from infected animals following stimulation with hookworm antigens was partially restored in the presence of antigen-presenting cells from uninfected hamsters. Analysis by fluorescence-activated cell sorting revealed that hookworm infection is associated with reduced percentages of both CD4(+) and surface immunoglobulin G-positive lymphocytes in the spleen and MLN cells. Splenocytes from infected hamsters also secreted more nitric oxide (NO) in culture than did those from naïve animals. Inhibition of NO secretion was associated with partial restoration of the proliferative capacity of splenocytes from infected animals in response to concanavalin A, suggesting a role for NO in mediating this effect. Together, these data demonstrate that hookworm infection is associated with impaired function of antigen-presenting cells and depletion of important lymphocyte subpopulations and also suggests a role for NO in parasite-induced immunosuppression.

Ancylostoma duodenale infection: a study of serum immunoglobulin G4 response to the excretory secretory antigen of adult worm.

The serum anti-Ancylostoma duodenale immunoglobulin (Ig) G4 antibody response to fraction III of the partially purified excretory secretory antigen of adult worm (Ad III ESA) was studied. The work included 60 patients with A. duodenale infection (GI), 40 patients with other parasitic infections (GII) and 30 apparently healthy parasite-free controls (GIII). Level of serum specific IgG4 was measured by indirect enzyme linked immunosorbent assay and compared with serum specific IgG, IgG 1, 2 & 3 subclass antibodies. Patients of GI had gastro-intestinal manifestations and symptoms suggestive of anaemia, and by investigations they had anaemia in 31.7% & eosinophilia in 100%. Measuring the intensity of A. duodenale infection, quantified as fecal egg counts, in patients of GI revealed that 60%, 30% & 10% had light, moderate, and heavy infections, respectively. The serum anti-Ad III ESA IgG & IgG 1-4 subclass antibodies were significantly elevated (P < 0.001) in patients of GI compared with GIII. Serum specific IgG4 was expressed in 100% of patients of GI at a significantly highly elevated level than IgGI (P < 0.01), IgG2 & IgG3 (P < 0.001). Specific IgG1 was expressed in 88.3% of patients of GI at a significantly elevated level (P < 0.001) than IgG2 & IgG3 which were expressed in 31.7% & 38.3%, respectively and elevated to a moderate extent. Serum specific IgG4 showed a 1.0, 1.1, 3.1 & 2.6-fold increase in detection rate of positive cases than IgG, IgG1, IgG2 & IgG3, respectively. The highest ability to differentiate between infected and healthy subjects was by serum specific IgG4 recording a discrimination coefficient of 9.4, while IgG, IgG1, IgG2 & IgG3 recorded 5.2, 6.3, 3.2 & 3.4, respectively. Serum specific IgG4 showed a significant positive correlation (r = 0.41, P < 0.001) with the intensity of A. duodenale infection, as was demonstrated by IgG & IgG3 (P < 0.01 & P < 0.05, respectively). Detection of serum anti-Ad III ESA IgG4 antibody recorded a 100% sensitivity that was significantly higher (P < 0.001) than IgG1, IgG2 & IgG3, but insignificantly different (P > 0.05) from IgG. Finally, serum specific IgG4 recorded a 100% specificity that was significantly higher than IgG, IgG2, IgG3 (P < 0.01) & IgG1 (P < 0.05). They showed cross-reactions with ascariasis, lymphatic filariasis and strongyloidiasis. The results are discussed.

[Iron-deficiency anemia related to ancylostoma duodenale infection among Ethiopian immigrants to Israel].

BACKGROUND: Among the diseases imported by the Ethiopian immigrants to Israel are many parasite infections. Hookworm infections, caused by the nematodes Necator americanus and Ancylostoma duodenale, involve the gastrointestinal tract, causing iron-deficiency anemia. OBJECTIVES: To evaluate the rate of iron-deficiency anemia related to hookworm infections among immigrants from an endemic area. METHODS: We retrospectively studied the medical records of all Ethiopians over the age of 18, who immigrated to Israel in the early 90's, in two primary care clinics of Clalit Health Services in Ofakim. RESULTS: Sixty patients (64%) had evidence of A. duodenale infection. The mean hemoglobin level was 11.92.3 g/dl in the Ancylostoma group and 13.81.6 g/dl in the control group (p = 0.0001). Analyzing the data according to the patient's sex revealed significant differences in the hemoglobin levels between the Ancylostoma group and the control group. Patients infected with A. duodenale had significantly lower mean corpuscular volume (MCV) and serum iron, and were likely to have eosinophilia and hypoalbuminemia. CONCLUSION: Among the Ethiopian immigrant population, Ancylostoma duodenale infection is a common cause of iron deficiency anemia. In young patients it should be ruled out before invasive and expensive investigations are performed.

T-lymphocyte subsets in patients with hookworm infection in Zaria, Nigeria.

Peripheral lymphocyte subsets CD3, CD4, CD8 were studied using monoclonal antibodies to determine the mechanism of immunosuppression observed in an earlier study with total T-cells using the sheep erythrocyte rosetting technique. The study was carried out in 37 Nigerian patients with hookworm infection (Necator americanus and Ancylostoma duodenale and 17 healthy Nigerians as controls. The leucocyte migration inhibition (LMI) test was also carried out to assess the functional integrity of the lymphocytes, while the hookworm status of the patients was assessed by the Stoll technique for measuring egg counts. The results of the T-cell studied showed that CD3 and CD4 cell percentages were significantly depressed in hookworm patients compared to controls (P<0.05). The difference between mean CD8 percentages and absolutes in hookworms patients and controls were not significant. Absolute CD3 and CD4 numbers were not significantly lowered compared to controls in the study and CD4/CD8 ratios were insignificantly reduced. Patients with polyparasitism including hookworm infection showed significant reduction in CD3 and CD4 cells when compared with controls and with patients with hookworm infection alone. The leucocyte migration inhibition response to purified protein derivative of M. tuberculosis (PPD) was significantly decreased in hookworm patients compared to controls, confirming that functionally cell mediated immunity is depressed in hookworm infection.

Epidemiology of human ancylostomiasis among rural villagers in Nanlin County (Zhongzhou Village), Anhui Province, China: II. Seroepidemiological studies of the age relationships of serum antibody levels and infection status.

Anti-hookworm antibody serologic responses were measured in residents of an Anhui provincial Chinese village where Ancylostoma duodenale is the predominant hookworm. Antibody responses were measured against either soluble infective third-stage larval (L3) or adult antigens. Immunoglobulins of the IgG class, especially IgG4 correlated with both the prevalence and intensity of A. duodenale hookworm infections. In contrast, there was an inverse correlation with IgM, but no correlation with IgA or IgE. Circulating IgG4 antibody responses might serve as a surrogate marker for active A. duodenale hookworm infection.

Enzyme-linked immunoelectrotransfer blotting analysis of human serologic responses to infective hookworm larval antigen.

OBJECTIVE: To explore the possibility of using specific antigens for immunodiagnosis of hookworm disease in endemic area. METHOD: Infective third-stage larvae of the canine hookworm, Ancylostoma caninum (A. caninum), were prepared as the source of antigen. Enzyme-linked immunoelectrotransfer blotting (EITB) was employed as an immunodiagnostic method. RESULTS: Two immunodominant bands of hookworm antigens (42 kDa and 55 kDa) were recognized by the sera of hookworm-infected patients (serum dilution 1:200; antigen centrifuged at 36,000 r/m for 20 minutes, but not by sera from negative controls. CONCLUSION: The 42 kDa and 55 kDa A. caninum antigens might be the specific antigens that could be used for immunodiagnosis of hookworm disease in endemic area.

Immunoglobulin G subclass antibodies against excretory/secretory antigens of Ancylostoma caninum in human enteric infections.

Most patients with proven or suspected enteric infection with the common hookworm of dogs, Ancylostoma caninum, produce immunoglobulin G (IgG) and IgE antibodies to an immunodominant excretory/secretory antigen (Ac68) of the parasite. These antibodies were detected in both enzyme-linked immunosorbent assay (ELISA) and Western blots; the Western blot to detect IgG antibodies to Ac68 was the most specific and sensitive. The subclasses of IgG of the antibody response to the parasite were analyzed using Western blots with anti-IgG subclass-specific monoclonal antibodies as marker systems in an attempt to further improve the specificity of the assay. Eight patients with confirmed enteric infections with A. caninum (positive controls) were tested; six had antibodies in all IgG subclasses against Ac68. Twenty sera from patients with suspected enteric infection with A. caninum (manifested as eosinophilic enteritis or unexplained abdominal pain with peripheral eosinophilia) were tested; 16 had total IgG antibodies to Ac68, while IgG1, IgG2, IgG3, and IgG4 responses were found in 11, 10, 9, and 12 of these sera, respectively. Small numbers of sera from groups of patients infected with other helminths and from healthy blood donors had various combinations of IgG, IgG1, IgG2, and IgG3 antibodies to Ac68, but none of these sera had IgG4 antibodies to Ac68. Sera from all nine patients with human hookworm infection had IgG, IgG1, IgG2, and IgG3 antibodies to Ac68 and eight of the nine were also positive for IgG4 antibodies. These results indicate the Western blot to detect IgG4 antibodies to Ac68 is the most reliable immunodiagnostic test yet described for enteric infection with A. caninum, although this test does not discriminate between infections with human and canine hookworms.

Pathogenicity of the hookworm, Ancylostoma pluridentatum, in a Florida panther (Felis concolor coryi) kitten.

We evaluated clinical signs and administered anthelmintic treatment to a wild-caught, captive Florida panther (Felis concolor coryi) kitten from Big Cypress National Preserve, Florida (USA) infected with the hookworm, Ancylostoma pluridentatum. Clinical signs observed included poor body condition, lethargy, and below normal red blood cell numbers, hemoglobin concentration, and packed cell volume, and elevated eosinophil numbers. In addition, a maximum of 936 Ancylostoma sp. eggs/g of feces were found on Day 11 of captivity. Following oral administration of 20 mg/kg pyrantel pamoate on Day 11, 26 A. pluridentatum were collected from the feces. Based on the resolution of clinical signs, cessation of egg shedding, and a return to normal hematologic values following anthelmintic treatment, we believe that infection with A. pluridentatum was the primary cause of the stressed conditions in the panther kitten.

Immunosuppression in Nigerians with hookworm infection.

T rosette cells results in patients with (Necator americanus and Ancylostoma duodenale) hookworm infection, divided into groups on the basis of the degree of anaemia (Group I had marked anaemia Hb 1-7, hypochromia ), Group II had moderate anaemia (Hb 8-11, hypochromia ++) and Group III had no signs of anaemia despite the underlying hookworm infection), were compared with results in 28 healthy controls who had no signs of other infection and excreting no ova of hookworm or other parasites. The level of sheep erythrocyte rosette-forming T-lymphocytes (SE-TL) in the patients (Group I:42 +/- 8%; Group II:45 +/- 11%; Group III:47 +/- 6%) was markedly reduced compared to controls (58 +/- 13%) (P < 0.001). The leucocyte migratory responses to purified protein derivative (P.P.D); were significantly (P < 0.05) reduced in all the subgroups as compared with the controls. The results support the hypothesis of the existence of immunosuppression in patients with hookworm infection.

Haematological changes in dogs infected with Ancylostoma duodenale.

A definite relation between the intensity of Ancylostoma duodenale and haematological changes in dogs were studied. There was a relation between the increase in egg count with the haemoglobin percentage. The present study reported haematological data collected during the acute phase of experimental infection with A. duodenale.

Occult enteric infection by Ancylostoma caninum: a previously unrecognized zoonosis.

BACKGROUND/AIMS: Human disease caused by the canine hookworm Ancylostoma caninum and a high incidence of eosinophilic enteritis have been reported from northern Queensland, Australia. The aim of this study was to evaluate patients with unexplained abdominal pain and a possible association with cryptic infections by A. caninum. METHODS: The clinical and demographic features of patients from this region with eosinophilic enteritis (group A1, n = 42), obscure abdominal pain associated with (group A2, n = 105) and without (group A3, n = 84) blood eosinophilia were reviewed and sera were tested against A. caninum excretory-secretory antigens by enzyme-linked immunosorbent assay (ELISA) and Western blot. Four additional patients, two with confirmed A. caninum, had hookworm infection. RESULTS: The level of dog ownership in these four groups was 79%-100%, higher than in the local population (P < 0.001). The ELISA tested positive in 71% of A1, 67% of A2, and 30% of A3, versus 8% in controls (P < 0.002). All cases tested were positive on Western blot versus 10% of controls (P < 0.0001). The ELISA values increased with chronicity and decreased during convalescence. CONCLUSIONS: We conclude occult human A. caninum infections are common and are characterized by eosinophilic enteritis and obscure abdominal pain with or without blood eosinophilia. The diagnosis can be confirmed by serology.

Weight loss, total and differential leucocytic counts in mice treated with alcohol during Ancylostoma caninum infection.

Females of Swiss albino mice were treated with various doses of alcohol and Ancylostoma caninum larvae. Mice which received 40% alcohol for 20 days showed a significant decrease in body weight and in the number of leucocytes. Eosinophilia was also observed in mice which were treated with 40% alcohol for 20 days.

[Parasitologic, clinical, hematologic and serologic findings in puppies after lactogenic infection with Ancylostoma caninum ERCOLANI 1859 (Ancylostomidae)]. / Parasitologische, klinische, hämatologische und serologische Befunde bei Welpen nach galaktogener Infektion mit Ancylostoma caninum ERCOLANI 1859 (Ancylostomidae).

The correlation between intensity of Ancylostoma caninum infections in bitches and the intensity of lactogenic infections and clinical signs of their puppies was investigated. On average, 825, 1,867 or 2,125 specimens were observed in litters of two bitches inoculated with 5,000, 10,000 or 20,000 third stage larvae (LIII) respectively, at the day of conception. Adverse effects of the infection on the growth and behaviour of the puppies were observed after onset of their second week of life: 11/27 puppies died during the fourth week. The body weight of puppies surviving 28 days was up to 750 g less than that of uninfected controls. Eosinophilia, erythroblastosis and microcytic, hypochromic anemia developed in all puppies during their first four weeks. The IFA test (LIII antigen) was positive for only 5/18 heavily infected puppies after uptake of colostrum.