Liesegang rings in a needle aspirate from a breast cyst

Ringlike structures of various sizes, resembling Liesegang rings (LRs), were found in the needle aspirate from a breast cyst in an elderly woman. They were characterized by a double-layer outer wall with striations and an amorphous central nidus. Immunohistochemical stains for calcium, iron, mucus, glycogen, amyloid, cytokeratin and epithelial membrane antigen yielded negative results. Since LRs can be mistaken for ova, larvae or parasites, it is important that cytologists be aware of their occurrence, albeit infrequent, in aspirated material to avoid a misdiagnosis.     

Ringlike structures in fine needle aspirates from the breast

Spherical ringlike structures of various sizes resembling Liesegang rings (LRs) are described in four fine needle aspirates (FNAs) from breast lesions over a period of 13 years. A characteristic finding in these structures was a distinct double layer outer wall with striations and an amorphous central nidus. Under polarized light they were non-refractile and no birefringence was noted in Congo red. Immunohistochemical stains for calcium, iron, mucus, glycogen, amyloid, cytokeratin and epithelial membrane antigen in all the cases were found to be negative. Since LRs can be mistaken for ova or parasites, their presence in aspirates of breast should be kept in mind to avoid misdiagnosis.     

Risk of contamination of human and agricultural environment with parasites through reuse of treated municipal wastewater in Riyadh, Saudi Arabia.

Two selected sites in the Riyadh metropolitan area were surveyed for the presence of human pathogenic parasites in treated municipal wastewater (TMWW). A total of 100 samples were collected from both sites at two different seasons, the winter and summer reason. The most common parasites seen were the larvae and adult of Strongyloides sp. There were few Ascaris lumbricoides ova, some of which were embryonated and possibly infective, while the highest frequency of Ascaris ova (100 +/litre) was found at site 2 h, the highest frequency of Strongyloides sp. larvae (36-72/litre) and Strongyloides sp. adult (100 +/litre) were found at sites 2 W and 1 W respectively. The variation between sites and seasonal fluctuations showed a significant difference in parasite per litre. High atmospheric temperatures in the Riyadh area seem to be lethal to most intestinal pathogens similarly the absence of protozoal cysts in the TMWW could be attributed to certain treatment processes and other environmental factors. Data obtained from this study will be valuable in planning for the control of parasitic diseases in particular and public health pathogens in general.     

Observations on the occurrence of crystal-like structures in nematode parasites of sheep and cattle

Waller P. J., Dash K. M. and Major G. W. 1979. Observations on the Occurrence of crystal-like structures in nematode parasites of sheep and cattle. Internationl Journal for Parasitology9: 147–151. Intracellular rod-like inclusions were found in a high percentage of inhibited fourth-stage larvae of H. contortus in sheep with naturally acquired infections. Artificial infections showed inclusions occurred in developing as well as inhibited fourth-stage larvae. The presence of these structures was associated with degenerative changes of the parasites and serial worm counts showed that larvae with large numbers of inclusions failed to persist within the host.Large refractive hexagonal crystals were observed in the gut lumen of fourth-stage and adult O. ostertagi acquired by previously worm-free lambs grazed on cattle pastures. The crystals tended to accumulate in the posterior gut region where they may cause gut blockage and lead to early mortality of the parasites in the abnormal host. No inclusions or crystals were observed in O. ostertagi from cattle or goats, or in O. circumcincta from sheep.     

Alternation of sleeping groves by yellow baboons (Papio cynocephalus) as a strategy for parasite avoidance

The primary locus of contact between free-living yellow baboons (Papio cynocephalus) in the Amboseli National Park of Kenya and the infective stages of their intestinal parasites is believed to be soil beneath sleeping trees contaminated by the baboons' own fecal emissions. In this report, we present evidence both that infective ova and larvae of intestinal parasites are found at very high densities in the soil beneath sleeping groves and that Amboseli baboons substantially reduce their contact with this reservoir of parasites by alternating periods of a few consecutive nights' use of any particular grove with much longer periods of avoidance of that grove. Although many factors other than the presence of parasite larvae also influence choice of sleeping groves, we propose as a working hypothesis that the temporal pattern of sleeping grove alternation shown by Amboseli baboons reflects a subtle behavioral strategy for parasite avoidance.     

Chronic wuchereriasis presenting as a vaginoperineal fistula: report of a case with aspiration cytologic diagnosis

BACKGROUND: Filariasis is often an occult disease with myriad presentations. Cytology has an established role in diagnosing clinically unsuspected cases. CASE: A 20-year-old female presented with recurring perineal ulcers, vaginal discharge and inguinal lymphadenopathy. Radiology revealed a vaginoperineal fistula. Fine needle aspiration (FNA) of the ulcer bed and smears prepared from the ulcer discharge showed an adult, gravid, female filarial worm and unsheathed larvae. Similar microfilariae were also seen in FNA smears from inguinal lymph nodes. The atypical clinical presentation and unusual parasite morphology posed initial difficulties in characterization of the microfilarial species. CONCLUSION: This case report highlights the morphologic clues to the dif ferential diagnosis offilarial species on cytologic specimens. Chronic wuchereriasis presenting as a vaginoperineal fistula has not been reported previously.     

Effect of a nuclear polyhedrosis virus on the relationship between Trichoplusia ni (Lepidoptera: Noctuidae) and the parasite, Hyposoter exiguae (hymenoptera: Ichneumonidae)

The effect of a nuclear polyhedrosis virus on the relationship between Trichoplusia ni and the parasite, Hyposoter exiguae, was investigated to determine if the virus could invade and multiply in the tissues of the parasites, if parasites which emerged from virus-infected T. ni larvae had normal emergence, fecundity, and longevity, and if the parasite could serve as a vector for the virus. Light microscopy revealed particles which appeared to be polyhedra within the lumen of the midgut of parasite larvae from virus-infected hosts. Transmission electron microscopy confirmed the presence of polyhedra and free virions within the midgut of the larvae. Polyhedra or free virions were never found within any parasite tissues. Parasite larvae within hosts exposed to virus before parasitization perished when their hosts died of virus infection. Parasite larvae in hosts exposed to virus after parasitization completed their development before their hosts died of virus infection. The proportion of parasites which survived increased as the time between host parasitization and host virus exposure increased. Parasite larvae which developed in hosts exposed to the virus soon after parasitization spent significantly less time in their hosts than did parasites which developed in noninfected hosts. There was no significant difference in time spent in the pupal stage, percent adult emergence, adult longevity with and without food and water, and fecundity of parasites which developed in virus-infected hosts and those which developed in noninfected hosts. Female parasites laid as many eggs in virus-infected hosts as they did in noninfected hosts. Sixty percent of the female parasites which oviposited in virus-infected hosts vectored infective doses of virus to an average of 6% of the healthy hosts subsequently exposed to them. None of the healthy host larvae exposed to male parasites which had been exposed to virus-infected host larvae became infected with the virus. Forty percent of the female parasites which developed in virus-infected hosts transmitted infective doses of the virus to an average of 65% of the healthy host larvae exposed to them. Ninety percent of the male parasites which developed in virus-infected hosts transferred infective doses of the virus to an average of 21% of the healthy host larvae exposed to them.     

SYSTEMATIC POSITION OF THE COPEPOD LERN AEOCERA CENTROPRISTI

The larvae of the lernaeocerid copepods are as yet very imperfectly known. To extend the knowledge of their morphology, the larvae of Lernaeocera centropristi Pearse, 1947, were re-examined. The larvae had been discovered on the gills of Centropristus striatus (L.) in Beaufort, North Carolina. Their adult stage is still unknown. The examination has shown that the larvae are almost identical with the known larvae of Lernaeocera , but also that they are equipped with additional armament at the base of the mouth-cone. Since the presence of this armament appears to be the main distinguishing feature between the larvae of the genera Lernaeocera and Lernaeenicus , and in view of other small distinctions, it is proposed that Lernaeocera centropristi Pearse, 1947, should be transferred to the genus Lernaeenicus , as L. centropristi (Pearse, 1947).     

Crystals and their taxonomic significance in some Verbenaceae

MATHEW, L. & SHAH, G. L., 1984. Crystals and their taxonomic significance in some Verbenaceae. The morphology, plant distribution and chemical composition of crystals in 35 tropical species in the family Verbenaceae have been investigated using light microscopy, SEM and an energy dispersive X-ray analyser.
Crystals may be solitary, aggregated or as conglomerates and crystal sand. Eleven kinds of solitary crystals and five aggregate or conglomerate forms have been observed. Their occurrence varies from species to species and sometimes from organ to organ in the same plant, suggesting that there might be taxonomic significance of crystal form and distribution.
Most of the crystals are compounds of a range of elements, silicon, calcium, potassium, iron, magnesium, aluminium, sulphur sodium and titanium in varying proportions. Calcium is the major component of solitary acicular crystals and silicon of other kinds of crystals.     

Liesegang rings in a dermoid cyst of the floor of the mouth. Report of a case with cytologic findings

BACKGROUND: Eosinophilic, ring-shaped structures, referred to as Liesegang rings (LRs), have been identified in aspirates from many organs, usually in relation to cystic or inflammatory lesions, but never before in relation to a dermoid cyst. CASE: LRs were intermingled with squamous cells in aspirates from a cyst of the floor of the mouth. The patient, a 23-year-old man, presented with a sublingual mass that had recently grown. Histopathologic study showed a ruptured dermoid cyst with mural foci of granulomatous foreign body reaction mixed with numerous ringlike structures, LRs. CONCLUSION: To the best of our knowledge, this is the first reported case of LRs in a dermoid cyst. Pathologists should be aware of this association when diagnosing cystic lesions.     

Cytological diagnosis of filarial infection in an endemic area where screening and prophylaxis is in place

P. Singh, Anshu and N. Gangane Cytological diagnosis of filarial infection in an endemic area where screening and prophylaxis is in place Objective: Filariasis has a worldwide distribution. However, the disease is often overlooked or misdiagnosed because of its unusual presentations and there may be false‐negative results in endemic areas. This study was carried out to assess the role of cytology in the demonstration of filarial parasites in an area where screening and prophylaxis are in place. Methods: This study was carried out in Wardha district in central India, which is endemic for filarial infection. A total of 9182 smears of cases undergoing cytological evaluation were routinely screened at the time of reporting for the presence of adult filarial worm, microfilarial larvae or their fertilized and unfertilized eggs, irrespective of their original clinical diagnosis. Results: Microfilariae were found incidentally in fine needle aspiration (FNA) smears of patients presenting with other clinical conditions. Smears of seven cases were found to be positive for microfilaria. Four cases presented with subcutaneous nodules in the upper arm, two cases were seen incidentally in axillary lymph node aspirates and microfilariae were found in the pleural fluid in one case. Another case was suspected to have breast cancer, but aspirates from the axillary area showed lymphatic obstruction considered to be due to filariasis. Conclusion: FNA cytology is an inexpensive, simple and easy procedure for detecting microfilariae. Detection of microfilariae may not be common even in endemic areas as screening and prophylaxis is routinely performed, and patients may show atypical presentations such as subcutaneous nodules. We believe that careful screening of FNAs, especially those from subcutaneous swellings, as well as serous fluids, is very important in a filariasis‐endemic zone.     

Fungal and oomycete parasites of Chironomidae,Ceratopogonidae and Simuliidae (Culicomorpha,Diptera)

Members of the families Chironomidae (chironomids or non-biting midges), Ceratopogonidae (ceratopogonids or biting midges) and Simuliidae (simulids or blackflies) are ubiquitous dipterans of the infraorder Culicomorpha. They are extremely diversified in ecological strategies. Their larvae play major roles in aquatic food webs as detritivores or predators, whereas their adults can be general predators (Chironomidae), hemolymphagous or hematophagous predators (Ceratopogonidae and Simuliidae) or pollinators. Both larval and adult stages are commonly infected by bacteria, viruses, protists, nematodes, true fungi and oomycetes. These phylogenetically diverse assemblages of microorganisms can simultaneously infect multiple species of chironomids, ceratopogonids and simulids, and each host may become trophically interrelated with other hosts by sharing their parasites. Here, we review the information on fungal and oomycete parasites of these dipteran groups with special reference to the natural regulation of host populations, the impact of parasitism in food webs, and the potential of these parasites as biocontrol agents.     

Nippostrongylus brasiliensis: effects of immunity on the pre-intestinal and intestinal larval stages of the parasite

Nippostrongylus brasiliensis: effects of immunity on the pre-intestinal and intestinal larval stages of the parasite. International journal for Parasitology4: 183–191. Migration of the pre-intestinal larval stages of N. brasiliensis was studied in rats undergoing either primary or challenge infections. In rats undergoing a primary infection, more than 67 percent of larvae successfully migrated from the skin to the oesophagus by 70 h after infection, and subsequently over 90 per cent of these larvae became established in the small intestine as sexually mature adults. In immune rats undergoing a second infection, 46 per cent of larvae completed migration to the oesophagus by 70 h and of these, only 1·6 per cent became established in the intestine to produce eggs. These inhibitory effects on the pre-intestinal and intestinal larval stages were even more pronounced in immune rats undergoing a third or fourth infection and in addition, there was a prolonged sojourn and substantial retention of larvae in their lungs. There was no evidence that the immune response had an adverse effect on oesophageal fourth stags larvae as these organisms (obtained from immune donors) were able to establish and develop to maturity when transferred per os to normal animals.Syngeneic transfer of immune mesenteric lymph node cells to normal recipients, caused expulsion of parasites from the intestine but failed to effect migration of pre-intestinal larval stages. The implications of these findings are discussed in the context of current knowledge of the mechanisms of immunity to helminths.     

The role of relatedness in structuring the social network of a wild guppy population

Parasite avoidance is increasingly considered to be a potential driving factor in animal migrations. In many marine and freshwater benthic fish, migration into a pelagic environment by developing larvae is a common life history trait that could reduce exposure to parasites during a critical window of developmental susceptibility. We tested this hypothesis on congeneric fish (family Galaxiidae, genus Galaxias) belonging to a closely related species complex sampled from coastal streams in southeastern New Zealand. Migratory Galaxias have larvae that migrate to pelagic marine environments, whereas the larvae of non-migratory species rear close to adult habitats with no pelagic larval phase. Both migratory and non-migratory fish are hosts to two species of skin-penetrating trematodes that cause spinal malformations and high mortality in young fish. Using generalized linear models within an Akaike information criterion and model averaging framework, we compared infection levels between migratory and non-migratory fish while taking into account body size and several other local factors likely to influence infection levels. For one trematode species, we found a significant effect of migration: for any given body length, migratory fish harboured fewer parasites than non-migratory fish. Also, no parasites of any kind were found in juvenile migratory fish sampled in spring shortly after their return to stream habitats. Our results demonstrate that migration spares juvenile fish from the debilitating parasites to which they would be exposed in adult stream habitats. Therefore, either the historical adoption of a migratory strategy in some Galaxias was an adaptation against parasitism, or it evolved for other reasons and now provides protection from infection as a coincidental side-effect.     

Parasites of recruiting coral reef fish larvae in New Caledonia

Recruiting coral reef fish larvae from 38 species and 19 families from New Caledonia were examined for parasites. We found 13 parasite species (Platyhelminthes: Monogenea, Cestoda and Trematoda) but no acanthocephalan, crustacean or nematode parasites. Over 23% of individual fish were infected. Didymozoid metacercariae were the most abundant parasites. We conclude that most of the parasites are pelagic species that become ‘lost’ once the fish larvae have recruited to the reef. Larval coral reef fish probably contribute little to the dispersal of the parasites of the adult fish so that parasite dispersal is more difficult than that of the fish themselves.     

Brugia pahangi: effects of duration of infection and parasite burden on lymphatic lesion severity, granulomatous hypersensitivity, and immune responses in jirds (Meriones unguiculatus)

The effects of Brugia pahangi infection duration and parasite burden on parasite-associated inflammatory and immune responses were determined over a 181-day period in jirds receiving from one to eight inoculations of infective larvae. Multiple infections did not produce a protective resistance to reinfection as determined by adult worm recovery at necropsy. Intralymphatic granulomatous lesions, lymph thrombi, were first seen at 48 days post initial inoculation (DPI). The numbers of lymph thrombi reached peak levels in singly inoculated jirds at 90 DPI and significantly decreased to low levels by 160 DPI. The ratio of lymph thrombi to adult worms recovered from the spermatic cord lymphatics followed a similar pattern. Sizes of renal lymph nodes, which drain lymphatics containing parasites, followed a temporal pattern of increase and decrease similar to that of lymph thrombi numbers. Peak granuloma areas around antigen-coated beads embolized in lungs were seen at 27 DPI. Granuloma areas around antigen-coated beads began to decrease after 69 DPI and reached sizes not significantly different from uninfected controls by 118 DPI. Multiple inoculations of infective larvae and increasing worm burdens did not affect the pattern of granulomatous response to antigen-coated beads. Eosinophilia of singly and multiply infected jirds peaked at 26 DPI. Eosinophilia of singly infected jirds returned to normal levels by 103 DPI but those of multiply infected jirds remained elevated until 160 DPI. Lymph node cell blastogenic responses to antigen were greater than those of splenocytes at all time intervals measured. However, significant differences in stimulation indexes between groups with different infection durations were not seen with either cell type. Antibody responses to somatic adult worm antigen as measured by ELISA reached near peak levels by 48 DPI and remained elevated for the course of the study in all infected jirds. The decrease in lymphatic lesion severity seen in chronically infected jirds temporally corresponds to the decrease in granulomatous reactivity measured around antigen-coated beads embolized in the lungs. This observation suggests that host and/or parasite factors associated with these two phenomena may be similar. Although these decreases may be the result of down-regulated immune responses, corresponding decreases in antibody levels and blastogenesis of lymphocytes stimulated by crude worm extracts were not observed in chronic infections.     

Trichinella spiralis: vascular recirculation and organ retention of newborn larvae in rats

The recirculation of Trichinella spiralis newborn larvae was studied in inbred AO rats. Newborn larvae collected after in vitro incubation of adult T. spiralis worms for 2 or 24 hr were injected into rats through the tail vein or hepatic portal vein. Blood samples from the femoral vein, hepatic portal vein, and abdominal aorta were collected at intervals from 1 min to 24 hr after larval injection. Newborn larvae of both ages (24 hr or 2 hr old) persisted in femoral vein blood for less than or equal to 5 hr after injection, but they could be detected in portal vein blood by 24 hr after injection. The injection of larvae into a tail vein or the portal vein did not influence the pattern of larval circulation, although there was a 1-5 min delay in newborn larval appearance time after injection into the portal vein. Transcapillary migration through tissue and back to the circulation was evident in the appearance of newborn larvae in the thoracic duct lymph up to 24 (occasionally 48) hr after tail vein injection of newborn larvae. During the course of a natural primary infection, no evidence for trapping of larvae in the mesenteric lymph node could be found despite direct larval migration through this organ. Injected newborn larvae were retained in the lungs, and small numbers could be recovered 24 hr after intravenous injection. We conclude that a proportion of newborn larvae recirculates within the vasculature for several hours; a smaller population extravasates but can reenter the circulatory system via the lymphatics. Furthermore, some newborn larvae are found in organs rich in capillaries up to 24 hr after their entry into the blood.     

Brugia malayi: transient transfection by microinjection and particle bombardment

To develop a method for the introduction of DNA into filarial parasites, several methods that have proven successful in other organisms were evaluated for their ability to transform Brugia malayi. Luciferase activity was detectable in embryos bombarded with gold particles coated with a construct consisting of a luciferase reporter gene under the control of the 5S rRNA intergenic spacer (SL promoter). Similar results were seen in adult parasites and infective larvae bombarded with this construct, or in adult female parasites microinjected with the plasmid. In similar experiments employing the SL promoter driving a green fluorescent protein (GFP) reporter, expression of the reporter was detectable in the intrauterine embryos of the microinjected adult parasites, and in the sub-cuticular tissues of biolistically transfected adult female parasites. A similar pattern of GFP expression to that seen in the SL promoter construct transfected parasites was noted in parasites transfected with constructs consisting of the upstream domain derived from an aspartyl aminoacyl tRNA synthetase gene of B. malayi. The ability to transfect B. malayi embryos may provide a foundation for studies of the regulation of gene expression and function in these organisms.     

Transmembrane regulation of intracellular calcium by a plasma membrane sodium/calcium exchanger in mouse ova

Regulation of cytoplasmic free calcium concentration ([Ca2+)]i) is a key factor for maintenance of viability of cells, including oocytes. Indeed, during fertilization of an ovum, [Ca2+]i is known to undergo oscillations, but it is unknown how basal [Ca2+]i or calcium oscillations are regulated. In the present study we investigated the role of the plasma membrane in regulating [Ca2+]i of metaphase II-arrested mouse oocytes (ova). Ova were collected from B6C3F1 mice treated with eCG (10 IU) and hCG (5 IU), and intracellular calcium was determined by means of fura-2. Extracellular calcium flux across the zona pellucida was detected noninvasively by a calcium ion-selective, self-referencing microelectrode that was positioned by a computer-controlled micromanipulator. Under basal conditions ova exhibited a calcium net efflux of 20.6 +/- 5.2 fmol/cm2 per sec (n = 69). Treatment of ova with ethanol (7%) or thapsigargin (25 nM-2.5 microM) transiently increased intracellular calcium and stimulated calcium efflux that paralleled levels of [Ca2+]i. The presence of a Na+/Ca2+ exchanger was indicated by experiments employing both bepridil, an inhibitor of Na+/Ca2+ exchange, and sodium-depleted media. In the presence of bepridil, a net influx of calcium was revealed across the zona pellucida, which was reflected by an increase in the [Ca2+]i. In addition, replenishment of extracellular sodium to ova that had been incubated in sodium-depleted media induced a large calcium efflux, consistent with the actions of Na+/Ca2+ exchange. Sodium/calcium exchange in mouse ova may be an important mechanism that regulates [Ca2+]i.     

Prevalence and abundance of Ixodes pacificus immatures (Acari: Ixodidae) infesting western fence lizards (Sceloporus occidentalis) in northern California: temporal trends and environmental correlates.

The prevalence and abundance of immature Ixodes pacificus ticks on western fence lizards (Sceloporus occidentalis) were examined in relation to time of year, host attributes (i.e., age, gender, and presence or absence of blood parasites), and 5 environmental characteristics, including topographic exposure and ground cover substrate, over a 2-year period in northern California. Lizards were infested with subadult ticks from early March until late July or early August, with peak median numbers of larvae and nymphs recorded in late April and early May of both years. Peak larval and nymphal abundances differed between years. The overall ratio of larvae to nymphs on adult male lizards was low, ranging from 0.80 in 1999 to 2.41 in 2000. Such intensive feeding of nymphs versus larvae on these lizards, which are reservoir-incompetent for Borrelia burgdorferi spirochetes, may explain previous observations of decreasing spirochetal infection prevalence from the nymphal to adult stage in northwestern California. Adult male lizards were more likely to be infested with nymphs and harbored greater abundances of larvae and nymphs than adult females. Lizards uninfected with blood parasites had more nymphs than infected lizards. The measured environmental characteristics could explain only a small percentage of the total variation observed in larval prevalence (22%) and in larval and nymphal abundance (12 and 3%, respectively).