Restoration of hookworm egg development after prolonged storage in stool suspension.

Hookworm infection is still prevalent in southern Thailand despite control measures. Hookworm eggs submerged for an extended period under water from rainfall or in latrines may not survive, but they may recover their ability to develop into infective larvae when exposed to atmospheric air. This study examined the survival of the hookworm eggs in stool suspension and the restoration of development capability after prolonged storage. In stool mass, eggs developed normally and yielded infective filariform larvae (FL) in 7 days. On the contrary, in 1:10 stool suspension, hookworm eggs were found to remain at the 4-8 cell stage; degenerated eggs were observed after 15 days of storage, and the number of degenerated eggs reached 80 % on day 30. Aeration of the suspension, or transferring to a Petri dish or agar plate, restored the capacity of eggs stored for up to 15 days to develop into FL; thereafter, the capacity declined sharply. Retardation of egg development under water or in stool suspension may be due to a lack of atmospheric air. Use of "night soil" from latrines as fertilizer may be one factor in maintaining hookworm transmission, as worm eggs can undergo normal development upon exposure to atmospheric air.

Rapid detection and identification of human hookworm infections through high resolution melting (HRM) analysis.

BACKGROUND: Hookworm infections are still endemic in low and middle income tropical countries with greater impact on the socioeconomic and public health of the bottom billion of the world's poorest people. In this study, a real-time polymerase chain reaction (PCR) coupled with high resolution melting-curve (HRM) analysis was evaluated for an accurate, rapid and sensitive tool for species identification focusing on the five human hookworm species. METHODS: Real-time PCR coupled with HRM analysis targeting the second internal transcribed spacer (ITS-2) of nuclear ribosomal DNA as the genetic marker was used to identify and distinguish hookworm species in human samples. Unique and distinct characteristics of HRM patterns were produced for each of the five hookworm species. The melting curves were characterized by peaks of 79.24±0.05°C and 83.00±0.04°C for Necator americanus, 79.12±0.10°C for Ancylostoma duodenale, 79.40±0.10°C for Ancylostoma ceylanicum, 79.63±0.05°C for Ancylostoma caninum and 79.70±0.14°C for Ancylostoma braziliense. An evaluation of the method's sensitivity and specificity revealed that this assay was able to detect as low as 0.01 ng/µl hookworm DNA and amplification was only recorded for hookworm positive samples. CONCLUSION: The HRM assay developed in this study is a rapid and straightforward method for the diagnosis, identification and discrimination of five human hookworms. This assay is simple compared to other probe-based genotyping methods as it does not require multiplexing, DNA sequencing or post-PCR processing. Therefore, this method offers a new alternative for rapid detection of human hookworm species.

Prevalencia de la infección por strongyloides stercoralis en la primera infancia: Orán, Salta, Argentina / Prevalence of strongyloides stercoralis infection in first childhood: Orán, Salta, Argentina

Un estudio enteroparasitológico, clínico y epidemiológico fue efectuado en 36 niños de 0 a 4 años (promedio 28 meses) de edad, que fueron llevados al Hospital San Vicente de Paul, Orán, Salta, en busca de asistencia médica. Las principales causas de consulta fueron bronquitis en el 52,8 por ciento de los casos, inapetencia en el 52,8 por ciento y palidez en el 27,8 por ciento . Las características generales de las muestras fecales fueron: formadas en el 44.4 por ciento de los casos y diarreicas en el 55,6 por ciento . Las frecuencias de formas diagnósticas correspondientes a helmintos y protozoos fue: S. stercoralis 30 (83,3 por ciento ), N. americanus 5(13,9 por ciento ), A. lumbricoides 7(19,4 por ciento ), T. trichiura 3(8,3 por ciento ), H. nana 5(13,9 por ciento ), G. intestinalis 10(27,8 por ciento ), I. belli 1(2,8 por ciento ) y E. coli 4(11,1 por ciento ). El estado nutricional encontrado en los niños, de acuerdo a la relación talla/peso fue: eutróficos 17(42,2 por ciento ), con desnutrición leve 9(20,0 por ciento ), con desnutrición moderada 9(25,0 por ciento ) y con desnutrición severa 1(2,8 por ciento ). Los valores medios del hematrocito y de la hemaglobina encontrados estuvieron por debajo de los establecidos como mínimos por O.M.S. En recuento de leucocitos y eosinófilos estuvo sobre los valores normales medios

Experimental and field studies on the viability of eggs, and the infectivity of third-stage larvae of the human hookworm Necator americanus recovered from the creek waters of the Niger Delta, Nigeria, and their epidemiological significance.

In a combined field and laboratory study, the eggs and 3rd-stage infective larvae of the human hookworm Necator americanus were recovered from the creek waters of the Niger Delta, Nigeria. They were tested for viability and infectivity. Eggs recovered from fresh water creeks showed a viability or hatching index of between 17.5 and 23.7%. Eggs recovered from brackish water showed a viability index of 18.8-21.4% in 'normal' cultures, and a maximum index of 6.3% when cultured in brackish water. The differences in the viability indices of these eggs were not significant (P greater than 0.05). Infective 3rd-stage larvae were recovered from both fresh creek water (205-258 L3/litre of water), and brackish water (45 L3/litre of water). Larvae hatched from brackish water eggs that were cultured in the brackish water were 20% infective by 3 days, but lost all infective potential by 7 days post-hatching. Larvae from fresh water eggs, cultured in fresh water and 'normal' laboratory cultures reached 50% infectivity in 3-5 days, losing potential infectivity in 11-15 days post-hatching. The infectivity patterns of larvae recovered directly from fresh and brackish waters did not differ significantly (P greater than 0.05) from the infectivity patterns of larvae recovered from laboratory cultures of eggs from all the experimental sources. Although larvae recovered directly from fresh and brackish waters survived in these respective media under laboratory conditions, there was a significant difference (P less than 0.05) in their survival patterns. The survival and infectivity rates of field larvae were considerably reduced over time relative to control larvae.(ABSTRACT TRUNCATED AT 250 WORDS)

Hookworm disease: host-pathogen biology.

The hallmark of hookworm infection, a common helminthic disease, is iron deficiency anemia. The development of anemia is dependent on the intensity of infection, the species of hookworm, and the ability of the host to resist infection and to maintain adequate stores of iron. When conditions are appropriate, the incidence of anemia caused by hookworm is high and has a significant economic impact since it results in a reduction of worker productivity. Loss of blood is caused by direct ingestion of red cells and by tissue trauma produced by worm attachment and feeding. This focal trauma may involve multiple villi and is characterized by local hemorrhage, tissue cytolysis, and neutrophilic response. Although focal intestinal lesions are apparent, their significance is questionable since diffuse mucosal changes are absent in intestinal biopsies of patients with heavy hookworm infection. Short-range control measures protecting against hookworm infection have not succeeded. Development of a vaccine against hookworm infection in humans is problematic since functional protective immunity in humans has not yet been demonstrated and no suitable animal model of hookworm infection in humans is available. At present, the most effective method of intervention appears to be supplementation of food staples with iron.

Skin penetration by Necator americanus larvae.

Skin penetration by Necator americanus larvae has been investigated in vitro. Larvae were able to penetrate completely human skin from both the epidermal and dermal directions; their passage through the epidermis was marked by cellular destruction. Removal of chloroform soluble skin lipids affected both the percentage of larvae invading and the percentage exsheathing. The larvae released an enzyme at about 37 degrees C, which showed peak activity against azocoll at 37 degrees C and pH 8. It is suggested that initial invasion is a mechanical process and that the enzyme is functional in passage through the germinal layers of the epidermis.

The infectivity of ingested adult hookworms.

Fourth-stage larvae and adults of Ancylostoma ceylanicum (dog strain) produced patent infections, with fertile ova, in hamsters to which they had been given in a drench. Similarly, 36-day-old (immature adult) Necator americanus given in a drench to hamsters also produced patent infections with fertile ova. The epidemiological significance of these experiments is discussed.

Pathogenicity of hookworms. The significance of population regression for the pathogenicity of hookworms.

A method is described to estimate the reduction of the hookworm population in the host during the infection period (population regression). A formula is proposed to estimate the average worm population during the infection period, on the assumption that a decreasing hookworm population causes a decrease in daily blood loss. Daily blood loss per worm was calculated for Ancylostoma caninum: 43 microliters; for A. ceylanicum: 14 microliters for A. braziliense: 5 microliters. Accordingly hookworms cause blood loss one half to two thirds smaller than formerly assumed.