Trichomonad infection in endemic and introduced columbids in the Seychelles

Island endemic avifaunas face many threats, including the now well-documented impacts of pathogens. The impacts of pathogens on the endemic Seychelles avifauna, however, have been little studied. The protozoan parasite Trichomonas gallinae has been shown to reduce survival and reproductive success of the endemic Pink Pigeon Columba mayeri on the nearby island of Mauritius. I investigated trichomonad infection prevalence and pathogenicity in endemic Seychelles Blue Pigeons, Alectroenas pulcherrima, and two introduced species of columbid, the Madagascar Turtle-dove, Streptopelia picturata, and the Barred Ground Dove, Geopelia striata, on the Seychelles island of Mahé during September-October 2007. I asked whether: 1) trichomonad infections occur in these species; 2) prevalence varies among species; and 3) birds show any signs of pathogenicity consistent with tricho-monosis. I use the results to assess the potential threat of this pathogen to A. pulcherrima. All three species were infected with trichomonads, and the overall prevalence was 27.5%. Alectroenas pulcherrima had higher prevalence (47.1%) than the two introduced species combined (24.3%). No infected individuals showed any signs of disease. These findings suggest that trichomonad parasites should be considered as a potential disease threat to the A. pulcherrima population.     

Detection of Dientamoeba fragilis in fresh stool specimens using PCR

Dientamoeba fragilis is a trichomonad parasite that causes human gastrointestinal disease. Currently microscopy is considered to be the gold standard for diagnosis of D. fragilis infection. However, this method is time-consuming and relatively insensitive. A PCR assay based on the small-subunit ribosomal RNA gene of D. fragilis for the specific detection of D. fragilis DNA in fresh unpreserved stool samples was developed. The D. fragilis PCR was positive in 29/31 samples with positive microscopy and did not cross-react with other protozoan parasites. The PCR protocol showed a specificity of 100% and a sensitivity of 93.5% and the entire procedure can be performed in one day.     

Entry and intracellular location of Mycoplasma hominis in Trichomonas vaginalis

The parasite Trichomonas vaginalis causes one of the most common non-viral sexually transmitted infections in humans. The coexistence of different sexually transmitted diseases in the same individual is very common, such as vaginal infections by T. vaginalis in association with Mycoplasma fermentans or Mycoplasma hominis. However, the consequences and behavior of mycoplasma during trichomonad infections are virtually unknown. This study was undertaken to elucidate whether mycoplasmas enter and leave trichomonad cells and if so how. M. hominis was analyzed in different trichomonad isolates and the process of internalization and the pathway within the parasite was studied. Parasites naturally and experimentally infected with mycoplasmas were used and transmission electron microscopy, cytochemistry and PCR analyses were performed. The results show that: (1) M. hominis enters T. vaginalis cells by endocytosis; (2) some mycoplasmas use a terminal polar tip as anchor to the trichomonad plasma membrane; (3) some trichomonad isolates are able to digest mycoplasmas, mainly when the trichomonads are experimentally infected; (4) some fresh virulent isolates are able to maintain mycoplasmas as cohabitants in the cell’s interior; (5) some mycoplasmas are able to escape from the vacuole to the trichomonad cytosol, and trichomonad plasma membrane budding suggested that mycoplasmas could leave the parasite cell.     

Pentatrichomonas hominis from beagle dogs--detection method, characteristics and route of infection

Pentatrichomonas sp. from the feces of beagles was cultured axenically, and identified as P. hominis. The culture medium used was slightly modified Diamond's medium supplemented with chicken liver extract and rifampicin. Based upon good proliferation after inoculating only a few organisms in this medium, a fecal examination method employing cultivation was developed. Resistance of the trichomonad against disinfectants and metronidazole was tested, and it was found that the protozoan was rather susceptible. After oral administration of the organism to mice and rats, all the treated animals were infected. Since two types of the trichomonad, moving and non-moving, were detected, the presence of any type resistant to standing or drying was ruled out. A possible route of trichomonad infection to beagles is discussed.     

Studies on Trichomonads. I. The Metabolism of Tritrichomonas foetus and Trichomonads from the Nasal Cavity and Gecum of Swine

SUMMARY. The metabolism of Tritrichomonas foetus (strain, BP-1) and trichomonads from the nasal cavity and cecum of swine was studied manometrically under similar experimental conditions. At pH 6.4, quantitative and qualitative differences were observed. The cecal (probably T. suis ) and nasal trichomonad used glucose, galactose, fructose, mannose, lactose, sucrose, raffinose, and trehalose. T. foetus used all except lactose and raffinose. All three were inhibited by iodoacetate and arsenite. T. foetus and the nasal form were significantly inhibited by fluoride and 8-hydroxyquinoline, whereas the cecal trichomonad was not. At varied pH, all failed to oxidize Krebs' cycle intermediates. The amounts of oxygen consumed by T. foetus and the nasal trichomonad in the presence of lactate and pyruvate were at levels similar to those with disaccharides; the cecal trichomonad was indifferent toward both substances. Anaerobically, lactate and pyruvate increased the evolution of gas by all three trichomonads. Aerobic acid formation was demonstrated for all three forms. Anaerobically, metabolic CO₂ and gas(es) that were not absorbed by KOH were evolved by all three. Pure oxygen was inhibitory to glucose utilization and stimulatory to the endogenous respiration of all trichomonads; the nasal form was affected the least.
The writer believes that the cecal trichomonad is different from T. foetus and the nasal trichomonad of swine. The relationship between the nasal trichomonad and T. foetus remains in doubt.     

Delayed Human Neutrophil Apoptosis by Trichomonas vaginalis Lysate

Neutrophils play an important role in the human immune system for protection against such microorganisms as a protozoan parasite, Trichomonas vaginalis; however, the precise role of neutrophils in the pathogenesis of trichomoniasis is still unknown. Moreover, it is thought that trichomonal lysates and excretory-secretory products (ESP), as well as live T. vaginalis, could possibly interact with neutrophils in local tissues, including areas of inflammation induced by T. vaginalis in humans. The aim of this study was to investigate the influence of T. vaginalis lysate on the fate of neutrophils. We found that T. vaginalis lysate inhibits apoptosis of human neutrophils as revealed by Giemsa stain. Less altered mitochondrial membrane potential (MMP) and surface CD16 receptor expression also supported the idea that neutrophil apoptosis is delayed after T. vaginalis lysate stimulation. In contrast, ESP stimulated-neutrophils were similar in apoptotic features of untreated neutrophils. Maintained caspase-3 and myeloid cell leukemia-1 (Mcl-1) in neutrophils co-cultured with trichomonad lysate suggest that an intrinsic mitochondrial pathway of apoptosis was involved in T. vaginalis lysate-induced delayed neutrophil apoptosis; this phenomenon may contribute to local inflammation in trichomoniasis.     

Dietary glycerol and adult access to water: effects on fecundity and longevity in the almond moth

The quality of food eaten by larval insects will affect traits such as gamete production, fat reserves, muscle bulk and body size in the adult. Moreover, larvae also depend on high moisture content in the diet for survival. The almond moth (Ephestia cautella) (W.) (Lepidoptera; Pyralidae) does not feed as an adult although it continues to drink water. We tested the idea that an almond moth could compensate for a low-water diet as a larva by increasing its water intake as an adult. We reared larvae on two different food sources with different moisture regimes; standard laboratory diet with glycerol (relatively wet) and standard diet without glycerol (relatively dry). Half the adult moths from each treatment were given water to drink before their first and only mating. Our results show that wet larval diets (i.e. containing glycerol) significantly decreased fecundity (total number of eggs laid and the proportion of hatched larvae), whilst it significantly increased male and female longevity. The interaction effect of water access for adult males and females was significant, independent of the glycerol in the larval diet. Longevity in females that were not presented with water as adults was slightly higher if mated with a male that had had access to water, suggesting a mating donation of water. However, females that received water as adults showed a decreased longevity if mated with a male who had also had access to water as an adult, indicating a negative effect of water if received by both males and females. In addition, when the larval diet included glycerol, increased number of eggs laid decreased female longevity, whilst an absence of glycerol in the larval diet resulted in low female longevity that was unlinked with fecundity. Glycerol is used in many artificial insect diets and the fact that it shows a strong effect on key life-history traits (reproductive output and longevity in this species), merits careful re-examination of its effects on these important traits in other laboratory models. We also discuss the possibility that larval diet can affect female reproductive decisions.     

A Realistic Appraisal of Methods to Enhance Desiccation Tolerance of Entomopathogenic Nematodes

Understanding the desiccation survival attributes of infective juveniles of entomopathogenic nematodes (EPN) of the genera Steinernema and Heterorhabditis, is central to evaluating the reality of enhancing the shelf-life and field persistence of commercial formulations. Early work on the structural and physiological aspects of desiccation survival focused on the role of the molted cuticle in controlling the rate of water loss and the importance of energy reserves, particularly neutral lipids. The accumulation of trehalose was also found to enhance desiccation survival. Isolation of natural populations that can survive harsh environments, such as deserts, indicated that some populations have enhanced abilities to survive desiccation. However, survival abilities of EPN are limited compared with those of some species of plant-parasitic nematodes inhabiting aerial parts of plants. Research on EPN stress tolerance has expanded on two main lines: i) to select strains of species, currently in use commercially, which have increased tolerance to environmental extremes; and ii) to utilize molecular information, including expressed sequence tags and genome sequence data, to determine the underlying genetic factors that control longevity and stress tolerance of EPN. However, given the inherent limitations of EPN survival ability, it is likely that improved formulation will be the major factor to enhance EPN longevity and, perhaps, increase the range of applications.     

Studies on Trichomonads. II. The Metabolism of a Trichomonas batrachorum-type Flagellate from the Cecum of Swine

SYNOPSIS. Certain aspects of the metabolism of a Trichomonas batrachorum -type flagellate from the cecum of swine were studied. This trichomonad (1) oxidized glucose, mannose, maltose, sucrose, and inulin, (2) was incapable of oxidizing Krebs' cycle intermediates, (3) possessed peroxide-splitting capabilities, (4) was inhibited by only iodoacetate and arsenite, and (5) formed acid(s) aerobically. Although there was no effect on oxygen uptake, pyruvate and lactate increased the anaerobic evolution of gas(es). In addition to CO₂, other gas, not absorbed by KOH, was produced anaerobically.
Compared with other porcine trichomonads, the metabolism of this small trichomonad resembles most closely that of the large cecal trichomonad, T. suis. However, the smaller trichomonad had a generally lower respiratory rate, a slightly lower optimal pH, and failed to oxidize fructose, galactose, lactose, raffinose, and trehalose.     

Pentatrichomonas hominis in laboratory-bred common marmosets

Trichomonadid protozoa have been found in the intestinal tracts of common marmosets (Callithrix jacchus). However, there is little information available on species identification and the pathogenicity of these trichomonads. In this study, we conducted a fecal survey of a common marmoset colony maintained as laboratory animals in Japan and identified the trichomonad species. Screening using a fecal smear examination revealed that 66% (58/88) of the marmosets had trichomonadid trophozoites in their feces. The trichomonads were found in both normal feces (31/49, 63%) and diarrhea (27/39, 69%), with no significant difference in frequency. The protozoa were identified as Pentatrichomonas hominis using morphological characters and the 100% identity of the nucleotide sequence of the partial 18S rRNA gene (297 bp). The intraspecific genetic variability between P. hominis from the marmosets in this study and P. hominis from other reported mammal hosts was ≤1% in the nucleotide sequence, including the internal transcribed spacer (ITS)-1, 5.8S rRNA gene, and ITS-2 (293 bp). P. hominis inhabits the large intestine of various mammalian hosts, including primates, and is considered nonpathogenic. These results suggest that P. hominis is transmitted among marmosets and other mammals but is not a primary cause of bowel disease in marmosets.     

First Report of Feline Intestinal Trichomoniasis Caused by Tritrichomonas foetus in Korea

Feline intestinal tritrichomoniasis by Tritrichomonas foetus was first recognized in USA in 1999 and has so far been reported from UK, Norway, Switzerland, and Australia, but not from the Far East Asian countries. In November 2008, 2 female and male littermate Siamese cats, 6-month old, raised in a household in Korea were referred from a local veterinary clinic with a history of chronic persistent diarrhea. A direct smear examination of fecal specimens revealed numerous trichomonad trophozoites which were isolated by the fecal culture in InPouch™ TF-Feline medium. A PCR testing of the isolate based on the amplification of a conserved portion of the T. foetus internal transcribed spacer (ITS) regions (ITS1 and ITS2) and the 5.8S rRNA gene, and the molecular sequencing of the PCR amplicons confirmed infection with T. foetus. This is the first clinical case of feline intestinal trichomoniasis caused by T. foetus in Korea.     

Molecular Characterization of Various Trichomonad Species Isolated from Humans and Related Mammals in Indonesia

Trichomonad species inhabit a variety of vertebrate hosts; however, their potential zoonotic transmission has not been clearly addressed, especially with regard to human infection. Twenty-one strains of trichomonads isolated from humans (5 isolates), pigs (6 isolates), rodents (6 isolates), a water buffalo (1 isolate), a cow (1 isolate), a goat (1 isolate), and a dog (1 isolate) were collected in Indonesia and molecularly characterized. The DNA sequences of the partial 18S small subunit ribosomal RNA (rRNA) gene or 5.8S rRNA gene locus with its flanking regions (internal transcribed spacer region, ITS1 and ITS2) were identified in various trichomonads; Simplicimonas sp., Hexamastix mitis, and Hypotrichomonas sp. from rodents, and Tetratrichomonas sp. and Trichomonas sp. from pigs. All of these species were not detected in humans, whereas Pentatrichomonas hominis was identified in humans, pigs, the dog, the water buffalo, the cow, and the goat. Even when using the high-resolution gene locus of the ITS regions, all P. hominis strains were genetically identical; thus zoonotic transmission between humans and these closely related mammals may be occurring in the area investigated. The detection of Simplicimonas sp. in rodents (Rattus exulans) and P. hominis in water buffalo in this study revealed newly recognized host adaptations and suggested the existence of remaining unrevealed ranges of hosts in the trichomonad species.     

Effects of sugars on the kinetics of drying and on the survival of partially dehydrated larvae of Anopheles mosquitoes

The ability to cryopreserve a stage of Anopheles mosquitoes would facilitate the development of strains incapable of transmitting malaria. Cryopreservation requires that the freezable water in cell systems be removed or rendered incapable of undergoing ice formation. The present study was concerned with the rate at which water is removed from lst instar larvae of Anopheles gambiae by air-drying, with the extent of dehydration that the larvae will tolerate, and with the effect of trehalose and sucrose on both drying kinetics and survival. Eighty-one percent of the larvae are water. Air-drying removes 90% of that water in approximately 20 min. Survivals after partial dehydration are highest if the larvae are rehydrated in 1/2x isotonic saline (0.13 osm); they are poorest if rehydrated in water or 0.13 osm sucrose. In the former, about 34% survive the removal of half the water, but next to none survive the loss of >70% initial water. Prior exposure to 0.2 M trehalose for as little as 1 min slows the drying rate and increases the tolerance of the larvae to dehydration. With 30-min exposure, 88% survive the loss of 50% of their water and 63% survive the loss of 75%. Protection is abolished with 0.4 M trehalose. The results are similar with sucrose. It is substantially reduced if sugar-exposed larvae are briefly washed with water prior to drying. The protection appears not to be related to the decreased drying rate. Rather it appears related, by an unknown mechanism, to the presence of sugar on the outer surface of the larvae.     

Hydrogenosome behavior during the cell cycle in Tritrichomonas foetus

The hydrogenosome is an unusual organelle found in several trichomonad species and other protists living in oxygen poor or anoxic environments. The hydrogenosome behavior in the protist Tritrichomonas foetus, parasite of the urogenital tract of cattle, is reported here. The hydrogenosomes were followed by light and transmission electron microscopy during the whole cell cycle. Videomicroscopy, immunofluorescence microscopy, and immunocytochemistry were also used. It is shown that the hydrogenosomes divide at any phase of the cell cycle and that the organellar division is not synchronized. During the interphase the hydrogenosomes are distributed mainly along the axostyle and costa, and at the beginning of mitosis migrate to around the nucleus. Three forms of hydrogenosome division were seen: (1). segmentation, where elongated hydrogenosomes are further separated by external membranous profiles; (2). partition, where rounded hydrogenosomes, in a bulky form, are further separated by a membranous internal septum and, (3). a new dividing form: heart-shaped hydrogenosomes, which gradually present a membrane invagination leading to the organelle division. The hydrogenosomes divide at any phase of the cell cycle. A necklace of intramembranous particles delimiting the outer hydrogenosomal membrane in the region of organelle division was observed by freeze-etching. Similarities between hydrogenosomes and mitochondria behavior during the cell cycle are discussed.     

Functional characterization of a novel plasma membrane intrinsic protein2 in barley

Water homeostasis is crucial to the growth and survival of plants. Plasma membrane intrinsic proteins (PIPs) have been shown to be primary channels mediating water uptake in plant cells. We characterized a novel PIP2 gene, HvPIP2;8 in barley (Hordeum vulgare). HvPIP2;8 shared 72–76% identity with other HvPIP2s and 74% identity with rice OsPIP2;8. The gene was expressed in all organs including the shoots, roots and pistil at a similar level. When HvPIP2;8 was transiently expressed in onion epidermal cells, it was localized to the plasma membrane. HvPIP2;8 showed transport activity for water in Xenopus oocytes, however its interaction with HvPIP1;2 was not observed. These results suggest that HvPIP2;8 plays a role in water homeostasis although further functional analysis is required in future.     

Variation in penaeid shrimp growth rates along an estuarine salinity gradient: Implications for managing river diversions

Freshwater inflows from river diversions may affect nekton populations by altering the salinity and temperature of estuarine waters. To investigate the influence of these environmental variables on the growth and survival rates of brown shrimp Farfantepenaeus aztecus and white shrimp Litopenaeus setiferus, we conducted field experiments in May and September 2007 to expose experimental animals to the range of different combinations of salinity and water temperature that commonly occur in an estuarine environment. Growth rates for shrimp held in mesocosms for approximately 7 days were compared among four locations and three treatments; locations were identified by the dominant marsh vegetation and distance from the Gulf of Mexico (low to high salinity: Intermediate, Brackish, Saline UE = Saline Up Estuary, Saline DE = Saline Down Estuary). At each location, the treatments were replicated four times and included shallow water with additional food, shallow water without food added, and deeper water (an attempt to expose animals to lower temperatures). Our experiments were designed to test the null hypothesis that shrimp growth and survival rates did not differ by location or treatment. Both brown shrimp and white shrimp grew more slowly at the Intermediate than higher salinity locations. Potential prey (benthic infauna) biomass was relatively low at both the Intermediate and Brackish locations in May, and both shrimp species consistently grew faster in mesocosms where food was added. We conclude that reduced growth in low salinity environments is likely due to the combined effects of increased metabolic costs and less food in these areas. River diversions that reduce estuarine salinities over a large portion of available habitat during peak recruitment periods may reduce overall growth rates and shrimp productivity in the affected areas.     

影响蝴蝶兰类原球茎耐脱水性的主要因素

为探讨蝴蝶兰(Phalaenopsis spp.)类原球茎(protocorm-like body,PLB)耐脱水性的主要影响因素,对PLB的平均粒重、含水率、脱水相对湿度、时间、温度、光周期与耐脱水性的关系进行了研究.结果表明,PLB的平均粒重与脱水后失水率、含水率、相对电导率、成活率呈显著或极显著相关.在较高湿度下...     

Loop-Mediated Isothermal Amplification Targeting Actin DNA of Trichomonas vaginalis

Trichomoniasis caused by Trichomonas vaginalis is a common sexually transmitted disease. Its association with several health problems, including preterm birth, pelvic inflammatory disease, cervical cancer, and transmission of human immunodeficiency virus, emphasizes the importance of improved access to early and accurate detection of T. vaginalis. In this study, a rapid and efficient loop-mediated isothermal amplification-based method for the detection of T. vaginalis was developed and validated, using vaginal swab specimens from subjects suspected to have trichomoniasis. The LAMP assay targeting the actin gene was highly sensitive with detection limits of 1 trichomonad and 1 pg of T. vaginalis DNA per reaction, and specifically amplified the target gene only from T. vaginalis. Validation of this assay showed that it had the highest sensitivity and better agreement with PCR (used as the gold standard) compared to microscopy and multiplex PCR. This study showed that the LAMP assay, targeting the actin gene, could be used to diagnose early infections of T. vaginalis. Thus, we have provided an alternative molecular diagnostic tool and a point-of-care test that may help to prevent trichomoniasis transmission and associated complications.     

Cryoprotective dehydration is widespread in Arctic springtails

Cryoprotective dehydration (CPD) is a cold tolerance strategy employed by small invertebrates that readily lose water by evaporation when subjected to sub-zero temperatures in the presence of ice. Until now, relatively few species have been investigated using methods by which CPD can be shown. In the present study we investigated the cold tolerance strategy of seven soil arthropod species from the high Arctic Spitzbergen, and compared water content and water loss, body fluid melting points (MP) and survival under cold and desiccating conditions. We tested the hypothesis that CPD is a commonly occurring cold hardiness strategy among soil arthropods. We found that four springtail species (Hypogastrura viatica, Folsomia quadrioculata, Oligaphorura groenlandica and Megaphorura arctica; Collembola) went through severe dehydration and MP equilibration with ambient temperature, and thus overwinter by employing CPD, whereas a beetle (Atheta graminicola) and one of the springtails (Isotoma anglicana) were typical freeze avoiding species over-wintering by supercooling. Desiccation tolerance of the red velvet mite (Neomolgus littoralis) was also investigated; very low water loss rates of this species indicated that it does not survive winter by use of CPD. All in all, the results of the present study confirm the hypothesis that CPD is an effective over-wintering strategy which is widespread within soil arthropods.     

三峡库区重庆段钉螺孳生条件

三峡工程的修建使得长江水生环境发生了变化,同时水库的阶段性蓄、放水使得阶段性淹没的江洲滩增加,更加有利于挺、浮水植物的繁殖,从而可能形成适合日本血吸虫(Schistosoma japonicum)的惟一中间宿主———钉螺(Oncomelania hupensis)的生长环境。本文在钉螺生活习性的基础上,结合数字高程模型(DEM)和相关的气象资料以及水库的蓄放水规律,从气候、淹没时间、植被、水库的蓄放水时间等角度对三峡库区钉螺孽生条件进行了分析。从气候上看,阶段性淹没区能够满足钉螺生长的需要;然而,水库冬季蓄水、夏季放水的周期与钉螺生长需求并不完全相同,尤其是冬季的高水位对钉螺生长十分不利。因此,水库的修建并不一定会造成钉螺的孽生和血吸虫的流行。