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Seal lice, unique among insects, show remarkable adaptability to the extreme conditions of the deep sea. Evolving with their seal and sea lion hosts, they have managed to tolerate hypoxia, high salinity, low temperature, and elevated hydrostatic pressure. Given the diving capabilities of their mammalian hosts, which can reach depths of hundreds to thousands of meters, our study examines the morphological variation among closely related seal lice species infesting hosts with different maximum diving depths. In particular, our research reveals a significant morphological difference between lice associated with regular and deep-diving hosts, where lice from deep-diving hosts tend to be rounder. This could be an adaptation to withstand the high hydrostatic pressures found in the deep ocean. The rounded shape optimizes the louse's ability to withstand external pressure by redistributing it over a larger ventral/dorsal plane. This in turn minimizes the internal energy required to support body deformations, thereby increasing the louse's resilience in the deep sea environment., (© 2024. The Author(s).)

Immature mosquitoes are thought to breathe only atmospheric air through their siphons despite reports of prolonged submerged survival. We studied the survival of last-instar larvae of Aedes aegypti fully submerged at different temperatures and measured the oxygen consumption from air and water-dissolved larvae and pupae of this species under different conditions. Larvae survived much longer than expected, reaching 50% mortality only after 58, 10, and 5 days at 15°, 25°, and 35 °C, respectively. Larval to pupa molt was only observed in larvae with access to air, whereas individuals kept submerged never molted. Although most of the oxygen was obtained from the air, larvae obtained 12.72% of their oxygen from the water, while pupae took only 5.32%. In both media, temperature affected the respiration rate of the larvae, with relatively close Q 10 values (1.56 and 1.83 for water and air, respectively). A similar pattern of O 2 consumption was observed in Ae. albopictus , whose larvae obtained 12.14% of their oxygen from the water. The detailed quantification of oxygen consumption by mosquito larvae showed that water-dissolved oxygen is not negligible and physiologically relevant, challenging the idea that mosquito larvae only breathe atmospheric oxygen.

Previous descriptions of Corynosoma bullosum (Linstow, 1892) show that specimens vary greatly in the proportions of different body structures, measurements of females and males, number of rows of hooks, and egg measurements, among other features. We redescribe this species from specimens found in southern elephant seal faeces from King George Island. We also provide a molecular characterization, in addition to 5.8S and internal transcribed spacer (ITS) existing sequences. We examined 41 elephant seals, and 30 adult acanthocephalans were found in 15 of them. The specimens were identified as belonging to the genus Corynosoma due to each having a tubular body with an inflated anterior part forming a thorny disk and the posterior part bearing somatic spines on the ventral surface, and genital spines surrounding the genital pore. Individual morphology corresponded to C. bullosum : large size, marked sexual dimorphism, and proboscis with 16-18 rows of spines with 11-15 spines per row. The molecular profile of three specimens of C. bullosum was analysed using 18S rDNA. We inferred phylogenetic relationships of the family Polymorphidae using maximum likelihood and Bayesian inference. We provide an updated morphological redescription for C. bullosum including electron microscopy photographs and molecular data. The 18S gene sequences showed low genetic variation and supported that C. bullosum is a sister to Corynosoma australe.

Through evolutionary time, seal lice have developed morphological, behavioral, and ecological adaptations to cope with the amphibious lifestyle of their hosts in a co-evolutionary process. Consequently, the dynamics of lice populations are determined by seals behavior. We aim to study the effects of host sex, age class, year, and sampling location, on the prevalence and mean abundance of Antarctophthirus carlinii, on Weddell seals (WS) Leptonychotes wedelli. The study was conducted at two sites in the Antarctic Peninsula, namely, Marambio/Seymour Island (MI) and the Danco Coast (DC). We collected lice from 71 WS: 33 from MI, during the reproductive season, and 38 from DC, during the molting season, between 2014 and 2017. According to our analyses, host age class and sex were the variables that affected prevalence levels of lice on WS. Whereas, age class, year, site, and sex affected lice mean abundance. Juveniles presented higher prevalence and mean abundance than adults, possibly acting as reservoirs for lice as they move through different colonies until they reach reproductive age. Concurrently, seals during molting season were more infested. Unlike nursing, during the molting season seals spend much time ashore forming mixed groups that favor both egg development and lice transmission.

Insects are the most evolutionarily and ecologically successful group of living animals, being present in almost all possible mainland habitats; however, they are virtually absent in the ocean, which constitutes more than 99% of the Earth's biosphere. Only a few insect species can be found in the sea but they remain at the surface, in salt marshes, estuaries, or shallow waters. Remarkably, a group of 13 species manages to endure long immersion periods in the open sea, as well as deep dives, i.e., seal lice. Sucking lice (Phthiraptera: Anoplura) are ectoparasites of mammals, living while attached to the hosts' skin, into their fur, or among their hairs. Among them, the family Echinophthiriidae is peculiar because it infests amphibious hosts, such as pinnipeds and otters, who make deep dives and spend from weeks to months in the open sea. During the evolutionary transition of pinnipeds from land to the ocean, echinophthiriid lice had to manage the gradual change to an amphibian lifestyle along with their hosts, some of which may spend more than 80% of the time submerged and performing extreme dives, some beyond 2000 m under the surface. These obligate and permanent ectoparasites have adapted to cope with hypoxia, high salinity, low temperature, and, in particular, conditions of huge hydrostatic pressures. We will discuss some of these adaptations allowing seal lice to cope with their hosts' amphibious habits and how they can help us understand why insects are so rare in the ocean.

Seals (Phocidae) undergo an annual cycle of moulting that implies hair regeneration, and in the case of southern elephant seals, it also involves the superficial strata of the epidermis. Therefore, surviving the moulting period is crucial for their obligate and permanent ectoparasites. Throughout evolutionary time, sucking lice (Echinophtiriidae) have developed morphological, behavioural and ecological adaptations to cope with the amphibious lifestyle of their hosts. Lepidophthirus macrorhini, the Southern elephant seal louse species, faces the additional challenge of surviving attached to the host during the moulting period. Since lice live on the skin, L. macrorhini has developed a unique survival strategy by piercing the skin of their host, thus keeping them protected from moulting. During fieldwork in Patagonia and Antarctica, skin samples with lice within were collected for histological analysis to assess whether these parasites caused damage to the host. Lice generate an inflammatory process in the host's dermis, and these lesions could alter the normal chemical and mechanical protective properties of the skin facilitating secondary infections. Further studies that analyse the potential pathogens in those skin lesions are necessary to properly assess the real impact of ectoparasites on their host health., (© 2021 The Royal Entomological Society.)

Lice from pinnipeds - sea lions, seals and walruses - are the only insects capable of surviving marine dives. Throughout their evolutionary history, they have adapted to tolerate hypoxia, high salinity, low temperature and, in particular, to tolerate conditions of high hydrostatic pressure. To understand the limits of the capacity of lice to survive during host deep dives, we conducted a series of controlled experiments in the laboratory. We collected lice from elephant seals and submitted the different life stages to high pressure conditions. Lice were first exposed to one of four hydrostatic pressures: 30, 80, 150 or 200 kg cm -2 They were then exposed a second time to higher or lower hydrostatic pressure conditions to test for the impact of the first experience, which could either be deleterious or trigger physiological adaption, allowing them a better tolerance to high pressure. We found that lice from elephant seals can tolerate hydrostatic pressures higher than 200 kg cm -2 (close to 200 atm), which is equivalent to 2000 m depth. Adults exhibited lower recovery times than nymphs after immersion at high hydrostatic pressure. Our findings show that lice have developed unique adaptations to endure extreme marine conditions. We discuss these extreme performances in relation to the morphological characteristics and physiological responses to diving in these insects., Competing Interests: Competing interestsThe authors declare no competing or financial interests., (© 2020. Published by The Company of Biologists Ltd.)

Lice from family Echinophthiriidae are of the few insects that have successfully colonized marine environment living as ectoparasites of pinnipeds, i.e., sea lions, seals, and the walrus. They have developed unique adaptations to cope with the amphibious lifestyle of their hosts. Because eggs do not survive underwater, lice could only reproduce when their host remains on pack ice enough time. Consequently, lice generations per year are limited by host haul-out behavior. The objective of this work is to study the effect of host sex and age class, and the annual variation on the prevalence and mean abundance of Antarctophthirus lobodontis in crabeater seals from the Antarctic Peninsula. During three consecutive field-seasons, we collected lice from 41 crabeater seals (23 females, 16 males, 2 indeterminate, being 24 adults, and 17 juveniles). We investigated this effect on the prevalence and mean abundance by a generalized linear model formulation in a Bayesian framework. According to the lowest Deviance Index Criterion model, sex host does not affect prevalence nor mean abundance. We found that juveniles present greater abundance and prevalence than adults, possibly due to foraging habits. They spent more time on the ice than adults in groups of dozens of animals. This behavior would favor both egg development and lice transmission. We do not find adult females with lice, which suggests that transmission of A. lobodontis should be horizontal. The high mean abundance of lice in 2014 could be associated with an unusual increase in Lobodon carcinophaga population, probably related to the pack-ice availability and zooplankton abundance.

[This corrects the article DOI: 10.1016/j.ijppaw.2017.08.002.].

Novel sequencing technologies are rapidly expanding the size of data sets that can be applied to phylogenetic studies. Currently the most commonly used phylogenomic approaches involve some form of genome reduction. While these approaches make assembling phylogenomic data sets more economical for organisms with large genomes, they reduce the genomic coverage and thereby the long-term utility of the data. Currently, for organisms with moderate to small genomes ($<$1000 Mbp) it is feasible to sequence the entire genome at modest coverage ($10-30\times$). Computational challenges for handling these large data sets can be alleviated by assembling targeted reads, rather than assembling the entire genome, to produce a phylogenomic data matrix. Here we demonstrate the use of automated Target Restricted Assembly Method (aTRAM) to assemble 1107 single-copy ortholog genes from whole genome sequencing of sucking lice (Anoplura) and out-groups. We developed a pipeline to extract exon sequences from the aTRAM assemblies by annotating them with respect to the original target protein. We aligned these protein sequences with the inferred amino acids and then performed phylogenetic analyses on both the concatenated matrix of genes and on each gene separately in a coalescent analysis. Finally, we tested the limits of successful assembly in aTRAM by assembling 100 genes from close- to distantly related taxa at high to low levels of coverage.Both the concatenated analysis and the coalescent-based analysis produced the same tree topology, which was consistent with previously published results and resolved weakly supported nodes. These results demonstrate that this approach is successful at developing phylogenomic data sets from raw genome sequencing reads. Further, we found that with coverages above $5-10\times$, aTRAM was successful at assembling 80-90% of the contigs for both close and distantly related taxa. As sequencing costs continue to decline, we expect full genome sequencing will become more feasible for a wider array of organisms, and aTRAM will enable mining of these genomic data sets for an extensive variety of applications, including phylogenomics. [aTRAM; gene assembly; genome sequencing; phylogenomics.]., (© The Author(s) 2017. Published by Oxford University Press, on behalf of the Society of Systematic Biologists. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Forty-one imperial shag chicks were sampled for lice during the breeding season of 2014 in Punta León, Argentina. We found 2 lice species, Pectinopygus turbinatus infesting the body and Piagetiella caputincisum present in the oral cavity of the birds. This constitutes the first host record for P. turbinatus and the first record for the continental Argentina for P. caputincisum . Ninety-three percent of the chicks were infested by at least one lice species. P . turbinatus was present in all of the lousy chicks, while P. caputincisum infested 84.2% of them. The mean intensity was 29.5 and the range 1-129. There was no difference in prevalence, mean intensity or mean abundance between louse species. However, we found differences among the pattern of infestation of each species. Imperial shag chicks were infested by their parents during their first days of life by P. turbinatus , mainly in nymphal stage and by P. caputincisum as adult lice. Our results showed differences among lice species that could be related to the restrictions that lice from seabirds faced during their life cycle.

The use of heat as a cue for the orientation of haematophagous insects towards hot-blooded hosts has been acknowledged for many decades. In mosquitoes, thermoreception has been studied at the molecular, physiological and behavioural levels, and the response to heat has been evaluated in multimodal contexts. However, a direct characterization of how these insects evaluate thermal sources is still lacking. In this study we characterize Aedes aegypti thermal orientation using a simple dual choice paradigm, providing direct evidence on how different attributes of heat sources affect their choice. We found that female mosquitoes, but not males, are able to discriminate among heat sources that are at ambient, host-range and deleterious temperatures when no other stimuli are present, eliciting a positive response towards host-range and an avoidance response towards deleterious temperatures. We also tested the preference of females according to the size and position of the sources. We found that females do not discriminate between heat sources of different sizes, but actively orientate towards closer sources at host temperature. Furthermore, we show that females cannot use IR radiation as an orientation cue. Orientation towards a host involves the integration of cues of different nature in distinct phases of the orientation. Although such integration might be decisive for successful encounter of the host, we show that heat alone is sufficient to elicit orientation behaviour. We discuss the performance of mosquitoes' thermal behaviour compared to other blood-sucking insects., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Despite the incredible success of insects in colonizing almost every habitat, they remain virtually absent in one major environment--the open sea. A variety of hypotheses have been raised to explain why just a few insect species are present in the ocean, but none of them appears to be fully explanatory. Lice belonging to the family Echinophthiriidae are ectoparasites on different species of pinnipeds and river otters, i.e. they have amphibious hosts, who regularly perform long excursions into the open sea reaching depths of hundreds of meters (thousands of feets). Consequently, lice must be able to support not only changes in their surrounding media, but also extreme variations in hydrostatic pressure as well as breathing in a low oxygen atmosphere. In order to shed some light on the way lice can survive during the diving excursions of their hosts, we have performed a series of experiments to test the survival capability of different instars of Antarctophthirus microchir (Phthiraptera: Anoplura) from South American sea lions Otaria flavescens, when submerged into seawater. These experiments were aimed at analyzing: (a) immersion tolerance along the louse life; (b) lice's ability to obtain oxygen from seawater; (c) physiological responses and mechanisms involved in survival underwater. Our experiments showed that the forms present in non-diving pups--i.e. eggs and first-instar nymphs--were unable to tolerate immersion in water, while following instars and adults, all usually found in diving hosts, supported it very well. Furthermore, as long as the level of oxygen dissolved in water was higher, the lice survival capability underwater increased, and the recovery period after returning to air declined. These results are discussed in relation to host ecology, host exploitation and lice functional morphology., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

As a part of an ongoing long-term study on the biology of pack-ice seals in Antarctica, we had the opportunity to collect lice from Weddell seals (Leptonychotes weddelli). We did not find the original description of this host-parasite association. Antarctophthirus ogmorhini had previously been reported as a parasite for the Weddell seal, but the information is, to a certain extent, confusing. During the development of the present study, we had access to literature concerning the presence of A. ogmorhini on this host, which, to our knowledge, was not determined in any of the previous works on this species. We compared lice collected from Weddell seals with A. ogmorhini obtained from the type host, the leopard seal (Hydrurga leptonyx), and we found that both species can be distinguished. The main differences are the characteristic pattern of chaetotaxy in the dorsal side of the head in lice from Weddell seals, the size and form of the pseudopenis, and the distribution and size of the fringe of setae surrounding the genital opening. Considering the conservative morphology, and ecological and evolutionary features of sucking lice, we proposed that lice from Weddell seals constitute a new species. In the present work, we described and illustrated adults of this new species collected from Weddell seals during the austral summer of 2014 at the Danco Coast, Antarctic Peninsula.

Among Anoplura, the family Echinophthiriidae includes those species that infest mainly the pinnipeds. Working with large hosts implies methodological considerations as the time spent in the sampling, and the way in that the animal is restrained. Previous works on echinophthiriids combined a diverse array of analyses including field counts of lice and in vitro observations. To collect lice, the authors used forceps, and each louse was collected individually. This implied a long manipulation time, i.e., ≈60 min and the need to physically and/or chemically immobilize the animal. The present work described and discussed for the first a sample technique that minimized the manipulation time and also avoiding the use of anesthesia. This methodology implied combing the host's pelage with a fine-tooth plastic comb, as used in the treatment of human pediculosis, and keeping the comb with the lice retained in a Ziploc® bag with ethanol. This technique was used successfully in studies of population dynamic, habitat selection, and transmission pattern, being a reliable methodology. Lice are collected entirely and are in a good condition to prepare them for mounting for studying under light or scanning electron microscopy. Moreover, the use of the plastic comb protects from damaging taxonomically important structures as spines being also recommended to reach taxonomic or morphological goals.

In this study, we document patterns of vertical transmission of the amphibious louse Antarctophthirus microchir (Echinophthiriidae) in pups of South American sea lion, Otaria flavescens, from Patagonia. Vertical transmission is fundamental for the long-term stability of A. microchir populations because only pups stay long enough (1 month) on land for the louse to reproduce. A total of 72 pups ≤7 days old from a single rookery were captured and examined for lice. Infection parameters and population structure of A. microchir did not differ among pups collected at the beginning, middle, and end of the reproductive season, suggesting that patterns of early vertical transmission are not affected by the increase of rookery size during this period. Over 60% of 1-day-old pups were infected with A. microchir, and recruitment increased in pups up to 3 days old and then leveled off. In 1-day-old pups, significantly more adults than nymphs were found, but the pattern was reversed in older pups. The number of first-stage nymphs was significantly smaller than that of second- and third-stage nymphs, as it was the number of males vs. females, particularly in 1-day-old pups. Three non-exclusive hypotheses could account for these patterns, i.e., recruitment merely reflects the population structure of A. microchir is cows; the relative ability of lice to pass from cows onto pups increases in advanced instars; and/or natural selection favors transmission of adults, especially females, because they accrue greater fitness. The importance of latter hypothesis should not be underestimated in a species with a tight reproductive schedule.

Background: Hepatitis B virus reactivation may occur in occult-infected carriers with haematological malignancies, whereas little data are available in patients undergoing chemotherapy for solid tumours., Aims: Evaluation of cancer patients undergoing chemotherapy to investigate occult hepatitis B virus infection and its clinical-virological outcome., Methods: Forty-four patients with solid tumours and without liver disease were prospectively enrolled and sampled before starting chemotherapy and between the second and third chemotherapy cycles (time points 1 and 2, respectively); 24 were also sampled 6 months after the end of chemotherapy (time point 3). At each time point, subjects were tested for liver biochemistry, hepatitis B serology and occult infection., Results: No sample tested positive for virus surface antigen. Twelve subjects (27.3%) were antibody positive to hepatitis B virus. Overall, occult infection was detected in 4 cases (9%), with positive HBV DNA at time points 1 and 2 (one case), at time point 1 only (one case), only at time points 2 and 3 (two cases), respectively. No occult-infected carrier experienced liver biochemistry flares and/or viral surface antigen positivity., Conclusions: Occult hepatitis B virus infection may occur in subjects with solid tumours, although the risk of its reactivation under chemotherapy appears to be very low., (Copyright © 2013 Editrice Gastroenterologica Italiana S.r.l. Published by Elsevier Ltd. All rights reserved.)

The members of the Family Echinophthiriidae (Phthiraptera: Anoplura) are unique among insects because they infest hosts with an amphibious lifestyle. During their evolution they developed morphological traits that are reflected in unique features. The SEM is a helpful tool to analyze them. Knowing in detail the external structure of these lice is the first step to understand the whole process that derived from the co-adaptation of lice and pinnipeds to the marine environment. For the first time, we studied the external structure of all stages of an echinophthiriid louse. The results are discussed in the light of their evolutionary, functional, and ecological implications., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Among Anoplura, the family Echinophthiriidae includes species that infest pinnipeds and otters. Previous evidence obtained from pinnipeds infested by echinophthiriids, specifically from seals, indicates that flippers are the preferred infestation sites, while lice from fur seals select areas in the pelage. We studied habitat selection of Antarctophthirus microchir on South American sea lion pups (Otaria flavescens Shaw, 1800) from Patagonia, Argentina, during the austral summer of 2009. We found a clear pattern of habitat selection: eggs are laid on the dorsal surface; nymphs 1 hatch there and then migrate to the belly, where they develop into adults and copulate; and then ovigerous females return to the dorsal surface. On the one hand, nymphs 1 are characterised by their low locomotory ability; therefore, the fact that they migrate as soon as they hatch suggests a clear pressure leading to microhabitat restriction. On the other hand, the described pattern of microhabitat selection seems to respond to the physiological requirements of each stage, which vary according to the physiological process considered, e.g. oviposition, morphogenesis, hatching and development. Accordingly, it appears that A. microchir would prefer the host's ventral area for development and copulation and the dorsal area for oviposition. However, the causes of this pattern are not clear, and many factors could be involved. Considering that sea lion pups periodically soak at high tides, and that prolonged immersion and very high humidity are known to be lethal for lice eggs, selecting the dorsal area would be advantageous for oviposition because it dries much faster. Furthermore, because humidity should be retained for longer periods on the ventral surface of the pup, wetter conditions on the sea lion would prevent desiccation of the nymphs in the very arid environment where O. flavescens breeds.

Antarctophthirus microchir was originally described from Phocarctos hookeri on the basis of 1 female and 1 male only. We redescribe adults and describe, for the first time, the 3 nymphal stages from specimens collected from Otaria flavescens from Patagonia, using light and scanning electron microscopy. The present material can be distinguished from other Antarctophthirus species by the presence of a fringe of setae on the back of the head, only present in Antarctophthirus trichechi and Antarctophthirus callorhini. However, A. trichechi also possess a prominent proboscis with large hooks, and A. callorhini presents less abundant and nonuniform abdominal scales in shape and size. Other differential features of A. microchir are the pattern of ovoid and uniform scales and longitudinal grooves in the surface of spines. Nymphal stage 1 differs from 2 and 3 mainly by the absence of scales and thorax without ventral spines or hairs. Nymphal stages 2 and 3 may be distinguished by the disposition of the occipital apophyses. Antarctophthirus microchir has been reported from 5 sea lion species from both hemispheres. Considering the conservative morphology, and ecological and evolutionary features of sucking lice, we raise the question of whether A. microchir from different sea lion hosts may represent a complex of cryptic species.

We analysed population dynamics of the louse Antarctophthirus microchir in pups of the South American sea lion, Otaria flavescens, at the Punta León rookery (Argentina) over a period of 2 years. A total of 136 pups were aged and marked at the beginning of the lactation period ashore, then sampled for lice at different times within 30 days. Sampling was restricted to the chest and belly, two sites where lice were especially abundant. This concentration on ventral areas might protect lice from thermal stress in the austral summer. Infestation patterns in pups 3 days old suggested that the potential for transmission increased from first nymphs to adults. Population trends of each instar with pup age, based on standardised values of abundance, were conserved between years, reflecting the basic dynamics of recruitment and reproduction. However, trends based on log-transformed abundances varied significantly between years; apparently, environmental conditions affected growth of lice populations differently each year. Stage-based deterministic models for population growth of A. microchir suggested generation times from 18 to 23 days. Accordingly, only 2 lice generations might be produced before pups start going to the sea. Shortening the cycle to accommodate a third generation might be risky, whereas a 2-generation cycle might at least result in larger females producing higher numbers of viable offspring.

Objective: The aim of this study was to describe a case of acute acalculous cholecystitis occurring in the course of primary Epstein-Barr virus (EBV) infection., Methods: The clinical features of the case were analyzed and compared to those of three other similar cases reported in the international literature., Results: All cases occurred in European females with cholestatic hepatitis, presented with gallbladder wall thickening, and recovered uneventfully without the need for surgical intervention., Conclusions: Acute acalculous cholecystitis may occur during the course of acute EBV infection, especially in patients with cholestatic hepatitis. Clinicians should be aware of the possible involvement of the gallbladder during EBV infection to avoid unnecessary invasive procedures or the overuse of antibiotics.

A Dot-ELISA is described that lends itself to the screening of large series of sera in relation to tetanus antibodies. A threshold of 0.06 I.U. per ml has been chosen as representing the protective tetanus antibody level. Two hundred and fifty-four sera were assayed by conventional ELISA, passive hemagglutination test and Dot-ELISA. The results obtained using concurrently these 3 techniques were in agreement with one another and permitted us to ascribe the same sera as tested by the various techniques to the same group of sera containing a non protective antitetanus antibody level and to the same group of sera-containing a protective level. It is suggested that Dot-ELISA should be used for the assessment of the tetanus immune status of populations.

An enzyme-linked immunosorbent assay using nitrocellulose strips (dot-ELISA) for the routine laboratory detection of IgG antibodies to mumps and varicella viruses is described. The virus antigens are dotted onto nitrocellulose strips, and the dotted strips are incubated with the sera to be tested. The bound antibodies are revealed using enzyme-labeled antihuman IgG antibodies. Reliable results are obtained when the assay is carried out at 37 degrees C. The reported data indicate that the dot-ELISA can reliably be used for the detection of IgG antibodies to mumps and varicella viruses in human sera.

A procedure for the routine and simultaneous laboratory detection of IgG antibodies produced in humans in the course of various infectious diseases is described. The procedure, based on dot-enzyme-linked immunosorbent assay (ELISA), used single nitrocellulose strips onto which several antigens were dotted in close proximity. Optimal conditions were specified that allowed the unequivocal and simultaneous detection of IgG antibodies specifically directed against Toxoplasma gondii, rubella virus, cytomegalovirus, and herpes simplex virus type 1 and type 2 antigens. This technique has proved to be simultaneously specific, sensitive, and reliable, and it has been applied to prenatal screening of sera from pregnant women. It is suggested that this technique should also be used for the screening of large numbers of sera under field trial conditions.

Eighty samples of human serum were tested for antibody to Toxoplasma gondii by enzyme-linked immunosorbent assay (ELISA) and indirect immunofluorescence (IIF). Development of ELISA suggested that accurate results could only be obtained with serial serum dilutions. ELISA and IIF tests showed a significant degree of correlation, but ELISA was found to be more sensitive: 17% of 24 sera showing negative IIF values showed slightly positive ELISA titres.

A modified dot-immunobinding assay is described for the rapid, specific and reliable screening of human sera for Toxoplasma gondii immunity status. This simple, rapid and sensitive assay proved to be a practical diagnostic technique since it can be performed in 3 hours employing a small amount of antigen (0.3 micrograms/microliters).

We describe a Dot-ELISA system for the rapid, specific and reliable assessment of a subject's antitetanus immune status. The conditions for Dot-ELISA were selected to give positive reactions only for sera with antitetanus antibody titers equal to or higher than 0.06 I.U./ml, considered to represent protection when using "in vitro" methods. This Dot-ELISA method could advantageously be applied to the monitoring of vaccination campaigns, as well as for assessing the antitetanus immune status of the wounded.

Many microorganisms and compounds of microbial origin exhibit immunomodulatory activities and have been extensively used in immunotherapy of experimental animal tumors and in patients with neoplasia. In this paper we describe the effect of the C. granulosum-derived P40 fraction on the growth and metastatization of the transplantable epithelioma T8 of Guèrin. Moreover, we evaluated the effect of P40 treatment on several depressed macrophage functions of tumor-bearing rats. In particular, the phagocytic and chemotactic activities of such cells were studied, as well as the antiviral intrinsic and extrinsic activities against HSV-1 and the anti-Toxoplasma gondii activity. All these functions were depressed in untreated tumor-bearing rats. Administration of a single intravenous injection of P40 fraction led to the restoration of all depressed macrophage activities to normal values. In particular, the possibility of restoring the antimicrobial activity of macrophages from tumor-bearing rats by treatment with this immunomodulator is of great concern when one considers the increasing incidence of opportunistic infections in immunocompromised hosts. Results are discussed in terms of both the possible mechanism of action of P40 and of its possible target cells.

A micro plate Enzyme-linked immunosorbent assay (ELISA), developed for detection of antibodies to Streptokinase, was used to analyze 80 human sera. The aim was to provide a simple method for antibody screening that required neither sophisticated equipment nor a high degree of technological skill. Preliminary results show that ELISA is specific, reproducible and sensitive. Comparative evaluation of the ELISA and the coagulum lysis test for antibodies against streptokinase show excellent correlation. We also evaluated the anti-Streptolysin O titer either by conventional method or ELISA. The sensitivity of ELISA for detecting anti-Streptolysin O antibodies was 3-4-fold higher than that of the hemolytic test. The sensitivity of ELISA for detecting anti-streptokinase antibodies was 10-20-fold higher than that of the coagulum lysis. The Enzyme-linked immunosorbent assay seems to offer some advantages over the more commonly used coagulum lysis test as far as sensitivity is concerned and potentially it is suggested that this method could be usefully employed clinically in human streptococcal infections.

A new ELISA test using the purified Listeria monocytogenes-derived antigen LM 84 Ag is described together with its application in the diagnosis of listeriosis. Fifty-seven human sera were titrated by the ELISA and the complement fixation method and compared for specificity and sensitivity. The problem of possible cross-reactions with other antigens is discussed.

In the last few years the interest about the influence on host/parasite relations exerted by antibiotics has increased. In this study we have studied the effect of ceftizoxime, a third generation cephalosporin, on some functional parameters of human macrophages and granulocytes. Ceftizoxime does not seem to exert any stimulatory effect on phagocytosis and chemotaxis, but at the same time it allows these cells to explicate their functions during an infective process. A separate series of experiments was designed in order to investigate the immunogenicity of ceftizoxime and its immunological cross-reactivity versus other beta-lactam antibiotics. The low ELISA title of ceftizoxime indicates its weak immunogenic power. Cross-reactivity was also very low (title 1:25) when ceftizoxime was tested against the other antibiotics.

The authors have carried out an epidemiologic research about the diffusion of antibody to hepatitis A antigen in the inhabitants of Ginostra, fraction of Stromboli, and Alicudi Islands (Eolie's arcipelago). We have examined by ELISA 86 human sera. We have detected a percentage of positivity about 82 and 70.6% for two populations. A critic examination of the results with parameters auxiliary (sex, age), showed significant differences of positivity about two different sexs and ages.

An indirect enzyme-linked immunosorbent assay, using antigen coupled to paper, has been adapted for the detection of Brucella melitensis antibodies. Optimum conditions were achieved by incubation of 1 ml of diluted serum with a single piece of paper coated with purified Brucella antigens for a period of one hour, and by addition of a goat anti-human enzyme conjugate antibody for one hour again. Under these conditions 80 human sera were examined and the results obtained were compared with Wright agglutination test.