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.

Secretion of a proteolytic anticoagulant by Ancylostoma hookworms.

Hookworms of the genus Ancylostoma secrete an anticoagulant that both inhibits the clotting of human plasma and promotes fibrin clot dissolution. This anticoagulant activity is attributable to a 36,000 dalton proteolytic enzyme. The protease can degrade fibrinogen into five smaller polypeptides that intrinsically have anticoagulating properties, covert plasminogen to a mini-plasminogen-like molecule, and hydrolyze a synthetic peptide substrate with specificity for elastolytic enzymes. It is hypothesized that the parasite uses this enzyme to prevent blood clotting while feeding on villous capillaries.

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.

The anticoagulant effects of the hookworm, ancylostoma ceylanicum: observations on human and dog blood in vitro and infected dogs in vivo.

Extracts of adult Ancylostoma ceylanicum prolonged the prothrombin time (PT) and partial thromboplastin time with kaolin ( PPTK ) of both human and dog plasmas in vitro. Excretory/secretory (E/S) products of these worms had similar effects while larval extract prolonged the PTTK only. Thus, the anticoagulant activities of this parasite are dependent upon the stage of the worm's life cycle. Collagen- and ADP-induced platelet aggregation were inhibited by adult and larval extracts. When the peripheral blood and bleeding times of dogs with varying worm burdens were examined, the only abnormality was shortening of the PTTK in the most heavily infected animals. Homogenates of dog small bowel subjacent to adult hookworms prolonged the PT of dog plasma and electron microscopical examination of this tissue revealed aggregation of platelets in blood venules without fibrin deposition. Thus, this study provides evidence that the anticoagulant properties of hookworms may have biological significance in infected animals.

Molecular basis of hyperlipidemia in golden hamsters during experimental infection with Ancylostoma ceylanicum (Nematoda:Strongylidae).

An infection of golden hamsters with Ancylostoma ceylanicum, a hookworm parasite, induced profound hyperlipidemia, particularly hypertriglyceridemia, and the effect was directly related to the degree of infection. A significant increase was also noticed in serum cholesterol and phospholipid levels. The appearance of lipoprotein-X, an abnormal low density lipoprotein, was detected in the serum of hookworm-infected animals. The hyperlipidemia was further characterized by an increase in very low density lipoproteins (VLDL) and low density lipoproteins (LDL) with a concomitant decline in high density lipoproteins (HDL). Decreased lipolytic activities, especially triglyceride lipase, in hepatic tissue and induction of lipolytic activities in intestine and adipose tissues indicated mobilization of fats from adipose and jejunum with a defective removal of triglyceride-rich lipoproteins in hepatic tissues. Accumulation of lipids in liver and depletion in adipose tissue supported these results. The derangement may have a significant effect on host parasite interaction and is an important pathophysiological feature occurring during experimental ancylostomiasis.

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.

Arrested development in Ancylostoma duodenale: course of a self-induced infection in man.

The course of self-induced infection with about 100 Ancylostoma duodenale larvae was followed by twice-weekly blood and stool examinations. After a small initial increase, no further change in eosinophil counts occurred until the 33rd week of infection when they began to increase sharply. Seven weeks later, eggs were first observed in stools. Thus, in this infection, the prepatent period was five times the expected. It is suggested that a strain of A.duodenale exists which either 1) has an inherent, abnormally long prepatent period or 2) has a prepatent period which is susceptible to change depending upon the environmental conditions experienced by the free-living larvae. In either case, the third stage larvae probably enter a hypobiotic state within the host which lasts for about 8 months.

alpha 1-Acid glycoprotein, alpha 1-antitrypsin, and ceruloplasmin in human intestinal helminthiases.

alpha 1-acid glycoprotein, alpha 1-antitrypsin, and ceruloplasmin levels were followed in 150 children from the rural tropics after arrival in the temperate zone. It is postulated that multiple intestinal helminthiases, especially ancylostomiasis, cause an acute phase type of reaction which is enhanced by immunization with T.A.B.-cholera vaccine.

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.

Ancylostoma caninum: a report on the peripheral eosinophilia in naive and immunized Swiss albino mice.

The number of peripheral eosinophils was counted in female Swiss albino mice on 1, 4, 9, 16 and 30 days after infection with various single (500, 1000 and 2000) and weekly repeated (500 + + 500 + 1000, 1000 + 1000 and 1000 + 1000 + 2000) doses of filariform Ancylostoma caninum larvae. The eosinophil response was significantly higher in infected than in uninfected mice and reached a peak on day 16 in naive and day 4 and 9 in immunized mice. Immunized mice were significantly more eosinophilic than the naive mice. An attempt has been made to correlate eosinophilia with immunity of mice to A. caninum larvae.

The response of captive rodents to the migration of Ancylostoma caninum larvae.

The distribution of the larvae of Ancylostoma caninum in tissues of captive wild rodents (Rattus rattus) and the leucocytic and behavioural responses of these rodents were studied after experimental oral infection. There was a wide distribution of larvae in tissues with a preponderance of the larvae in the skeletal muscles of the anterior part of the body in older infection. The rats responded by an elevation of total leucocytes and eosinophils in blood, alternation of locomotory activity and behavioural dominance that may have a correlation with predation and epidemiology of A. caninum in a sylvatic setting.

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.

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.

Investigation of the haemolytic effects of Ancylostoma ceylanicum: observations on infected dogs in vivo and human and dog blood in vitro.

Investigations into the probable role of haemolysins in the causation of hookworm anaemia have been undertaken in living infected dogs. Secondly, the effects of living hookworms and various worm products on erythrocytes in vitro have been assessed. In dogs infected with varying numbers of A. ceylanicum, severe microcytic anaemia developed in the most heavily infected animals six weeks after infection. Erythrocytes from the latter animals showed significantly greater autohaemolysis in the presence of added glucose. When serum bilirubin and methaemalbumin, plasma haemoglobin, urinary urobilinogin and osmotic fragility of their red cells were measured, however, no evidence of haemolysis was detected. Erythrocytes from these animals appeared normal under scanning electron microscopy. In in vitro studies varying concentrations of adult worm extract had no effect on the haemolysis of either dog or human erythrocytes in the presence or absence of glucose nor on their mechanical fragility. There was no increase in 51Cr release from dog or human labelled red cells when incubated with either adult worm extract or excretory/secretory products of worms. Living adult worms caused an increase in 51Cr release from human but not dog labelled erythrocytes. Thus, the role of haemolysins in the genesis of hookworm anaemia is minimal.

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.

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.

[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.