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In response to an outbreak of Crimean-Congo hemorrhagic fever in western Afghanistan, we measured immunoglobulin G seroprevalence among household members and their animals. Seroprevalence was 11.2% and 75.0% in humans (n = 330) and livestock (n = 132), respectively. Persons with frequent exposure to cattle had an elevated risk of being immunoglobulin G positive. [ABSTRACT FROM AUTHOR]

Abstract Background Lower respiratory tract infections (LRTI) are responsible for a considerable number of deaths among children, particularly in developing countries. In Egypt and the Middle East region, there is a lack of data regarding the viral causes of LRTI. In this study, we aimed to identify the relative prevalence of various respiratory viruses that contribute to LRTIs in young children. Although, nucleic acid-based methods have gained importance as a sensitive tool to determine the viral infections, their use is limited because of their prohibitive cost in low-income countries. Therefore, we applied three different laboratory methods, and presented the different virus prevalence patterns detected by each method. Methods We collected nasopharyngeal aspirate samples, demographic data and, clinical data from 450 children under five years of age who presented with LRTI at Abou El Reesh hospital in Cairo during a one-year period. To identify the viral causes of the LRTI we used direct fluorescence assay, real-time reverse-transcriptase polymerase chain reaction (rt-RT-PCR), and shell vial culture. We tested for eight major respiratory viruses. Results Two hundred sixty-nine patients (59.9%) had a viral infection, among which 10.8% had a co-infection with two or more viruses. By all three methods, respiratory syncytial virus (RSV) was the most predominant, and parainfluenza virus type 2 (HPIV-2), influenza B virus (FLUBV) were the least predominant. Other viral prevalence patterns differed according to the detection method used. The distribution of various viruses among different age groups and seasonal distribution of the viruses were also determined. Conclusions RSV and human adenovirus were the most common respiratory viruses detected by rt-RT-PCR. Co-infections were found to be frequent among children and the vast majority of co-infections were detected by nucleic acid-based detection assays.

An outbreak of nonspecific febrile illnesses occurred among U.S. Army troops in September 2007 at a remote, newly established, rural-situated patrol base, south of Baghdad, Iraq. Soldiers displayed an acute flu-like syndrome with symptoms of fever, headache, malaise, and myalgia. A total of 14 cases was identified and treated presumptively as query fever. Subsequent convalescent serum specimens confirmed 13 (92.9%) positive for sandfly Sicilian virus and 3 (21.4%) positive for Coxiella burnetii, with two positive for both. One sandfly Sicilian virus case tested positive for Brucella spp. This outbreak emphasizes the potential for multiple simultaneous disease exposures to endemic diseases in nonindigenous military personnel at remote military locations in Iraq. Recommendations include increased theater disease surveillance, medical training, and vector control. [ABSTRACT FROM AUTHOR]

Rodents serve as reservoirs and/or vectors for several human infections of high morbidity and mortality in the tropics. Population growth and demographic shifts over the years have increased contact with these mammals, thereby increasing opportunities for disease transmission. In Africa, the burden of rodent-borne diseases is not well described. To investigate human seroprevalence of selected rodent-borne pathogens, sera from 657 healthy adults in ten rural communities in Ghana were analyzed. An in-house enzyme-linked immunosorbent assay (ELISA), for immunoglobulin G (IgG) antibodies to Lassa virus was positive in 34 (5%) of the human samples. Using commercial kits, antibodies to hantavirus serotypes, Puumala and Dobrava, and Leptospira bacteria were detected in 11%, 12% and 21% of the human samples, respectively. Forty percent of residents in rural farming communities in Ghana have measurable antibodies to at least one of the rodent-borne pathogens tested, including antibodies to viral hemorrhagic fever viruses. The high seroprevalence found in rural Ghana to rodent-borne pathogens associated with both sporadic cases and larger disease outbreaks will help define disease threats and inform public health policy to reduce disease burden in underserved populations and deter larger outbreaks., Competing Interests: Dr. Nimo-Paintsil, Dr. Mohareb, Dr. Schoepp, CDR Letizia, and CAPT Kronmann are military service members or employees of the U.S. Government. This work was prepared as part of their official duties. Title 17 U.S.C. §105 provides that ‘Copyright protection under this title is not available for any work of the United States Government.’ Title 17 U.S.C. §101 defines a U.S. Government work as a work prepared by a military service member or employee of the U.S. Government as part of that person’s official duties. This does not alter our adherence to PLOS ONE policies on sharing data and materials.

Several Hantaviruses cause hemorrhagic fever with renal syndrome (HFRS) in Europe: Dobrava-Belgrade virus (DOBV), Puumala, Saaremaa, Sochi, and Seoul virus. Although HFRS is endemic in Bulgaria, genome sequences of hantaviruses have never been detected in wild rodents. To identify rodent reservoirs, a total of 691 rodents from three endemic regions were trapped in 2011-2012 and screened by TaqMan RT-PCR for detection of hantaviral genomic RNA. Partial small (S) and/or large (L)-segment sequences were recovered from six Apodemus mice: five of the species A. flavicollis and one A. agrarius. Phylogenetic analysis revealed that all recovered sequences belonged to DOBV. On the phylogenetic trees, the novel Bulgarian hantavirus sequences clustered together with sequences of established previously DOBV variants recovered from Bulgarian HFRS patients and also with variants found in wild rodents trapped in Slovenia, Greece, and Slovakia. One of the novel Bulgarian DOBV S-sequences from A. agrarius was related closely to DOBV sequences recovered from A. flavicollis, suggesting a spillover of DOBV from its natural host to A. agrarius mice. The results of this study confirmed the circulation of DOBV in wild rodents in Bulgaria. The complexity of the epidemiological situation in the Balkans requires further studies of hantaviruses in rodent hosts and human HFRS cases., (© 2014 Wiley Periodicals, Inc.)

Introduction: Zoonotic diseases are an important cause of human morbidity and mortality. Animal populations at locations with high risk of transmission of zoonotic pathogens offer an opportunity to study viral and bacterial pathogens of veterinary and public health concern., Methods: Blood samples were collected from domestic and imported livestock slaughtered at the Muneeb abattoir in central Egypt in 2009. Samples were collected from cattle (n=161), buffalo (n=153), sheep (n=174), and camels (n=10). Samples were tested for antibodies against Leptospira spp. by a microscopy agglutination test, Coxiella burnetii by enzyme immunoassay, Brucella spp. by standard tube agglutination, and Rift Valley Fever virus (RVFV), Crimean-Congo hemorrhagic fever virus (CCHFV), sandfly fever Sicilian virus (SFSV), and sandfly fever Naples virus (SFNV) by enzyme-linked immunosorbent assay., Results: Antibodies against Leptospira spp. were identified in 64 (40%) cattle, 45 (29%) buffalo, 71 (41%) sheep, and five (50%) camels; antibodies against C. burnetii in six (4%) buffalo, 14 (8%) sheep, and seven (70%) camels; and antibodies against Brucella spp. in 12 (8%) cattle, one (1%) buffalo, seven (4%) sheep, and one (10%) camel. Antibodies against RVFV were detected in two (1%) cattle and five (3%) buffalo, and antibodies against CCHFV in one (1%) cow. No antibodies against SFSV or SFNV were detected in any species., Discussion: RESULTS indicate that livestock have been exposed to a number of pathogens, although care must be taken with interpretation. It is not possible to determine whether antibodies against Leptospira spp. and RVFV in cattle and buffalo are due to prior vaccination or natural exposure. Similarly, antibodies identified in animals less than 6 months of age may be maternal antibodies transferred through colostrum rather than evidence of prior exposure. RESULTS provide baseline evidence to indicate that surveillance within animal populations may be a useful tool to monitor the circulation of pathogens of veterinary and public health concern in Egypt.

Little is known about the occurrence of chikungunya fever in the Eastern Mediterranean Region of the World Health Organization (WHO). In January 2011, the Ministry of Public Health and Population (MoPH&P) of Yemen reported to WHO an increasing number of "dengue-like" acute febrile illnesses of unknown origin from one of its coastal governorates. An epidemiological investigation was conducted in Al-Hudaydah governorate between 23 and 26 January 2011 by a joint team of WHO, the MoPH&P of Yemen and the U.S. Naval Medical Research Unit (NAMRU-3) in Cairo, Egypt. The investigation led to the detection of an outbreak of chikungunya in Yemen which was the first time ever from any of the 22 countries in the Eastern Mediterranean Region of WHO. Appropriate public health control measures were strengthened following the investigation, and the outbreak was contained. This paper provides a short description of the outbreak and its epidemiological characteristics and highlights the important lessons that were learned for early detection and control of chikungunya in countries where competent vectors for transmission of the virus exist., (Copyright © 2014 Ministry of Health, Saudi Arabia. Published by Elsevier Ltd. All rights reserved.)

Crimean-Congo haemorrhagic fever (CCHF) is a tick-borne zoonotic disease. Over the past decade, CCHF cases in humans have emerged in Turkey and reemerged in the Balkan countries, Ukraine and Tajikistan. Occupational contact with infected livestock has been recognized as a common cause of the disease. A cross-sectional seroprevalence study in livestock was conducted in farming communities of an endemic area in Bulgaria, southeastern Europe. Overall, 72% of the tested animals were positive for IgG antibodies to CCHF virus. By the time the animals were one-year old almost 50% had serologic evidence of CCHF infection, and by two years already 80% of them had been infected. The data obtained in this study reflect current situation of CCHF virus infection among livestock in Bulgaria. The results showed active CCHF virus circulation that poses risk for humans to be infected during contacts with animals and requires public health awareness.

A new human coronavirus (CoV), subsequently named Middle East respiratory syndrome (MERS)-CoV, was first reported in Saudi Arabia in September 2012. In response, we developed two real-time reverse transcription-PCR (rRT-PCR) assays targeting the MERS-CoV nucleocapsid (N) gene and evaluated these assays as a panel with a previously published assay targeting the region upstream of the MERS-CoV envelope gene (upE) for the detection and confirmation of MERS-CoV infection. All assays detected ≤10 copies/reaction of quantified RNA transcripts, with a linear dynamic range of 8 log units and 1.3 × 10(-3) 50% tissue culture infective doses (TCID50)/ml of cultured MERS-CoV per reaction. All assays performed comparably with respiratory, serum, and stool specimens spiked with cultured virus. No false-positive amplifications were obtained with other human coronaviruses or common respiratory viral pathogens or with 336 diverse clinical specimens from non-MERS-CoV cases; specimens from two confirmed MERS-CoV cases were positive with all assay signatures. In June 2012, the U.S. Food and Drug Administration authorized emergency use of the rRT-PCR assay panel as an in vitro diagnostic test for MERS-CoV. A kit consisting of the three assay signatures and a positive control was assembled and distributed to public health laboratories in the United States and internationally to support MERS-CoV surveillance and public health responses.

We used a seroepidemiologic study to estimate Q fever (Coxiella burnetii) seroprevalence, seroincidence, and risk factors for seroconversion in two deployed military populations in 2005. The first study group resided in an area with a known Q fever outbreak history (Al Asad, Iraq). Of this population, 7.2% seroconverted for an incidence rate of 10.6 seroconversions per 1,000 person-months. The second population included personnel transiting through Qatar on mid-deployment leave from southwest/central Asia. In this group, we found 2.1% prevalence with 0.92 seroconversions per 1,000 person-months. However, no significant risk factors for Q fever seroconversion were found in either population.

Surveillance for influenza A viruses in wild birds has increased substantially as part of efforts to control the global movement of highly pathogenic avian influenza A (H5N1) virus. Studies conducted in Egypt from 2003 to 2007 to monitor birds for H5N1 identified multiple subtypes of low pathogenicity avian influenza A viruses isolated primarily from migratory waterfowl collected in the Nile Delta. Phylogenetic analysis of 28 viral genomes was performed to estimate their nearest ancestors and identify possible reassortants. Migratory flyway patterns were included in the analysis to assess gene flow between overlapping flyways. Overall, the viruses were most closely related to Eurasian, African and/or Central Asian lineage low pathogenicity viruses and belonged to 15 different subtypes. A subset of the internal genes seemed to originate from specific flyways (Black Sea-Mediterranean, East African-West Asian). The remaining genes were derived from a mixture of viruses broadly distributed across as many as 4 different flyways suggesting the importance of the Nile Delta for virus dispersal. Molecular clock date estimates suggested that the time to the nearest common ancestor of all viruses analyzed ranged from 5 to 10 years, indicating frequent genetic exchange with viruses sampled elsewhere. The intersection of multiple migratory bird flyways and the resulting diversity of influenza virus gene lineages in the Nile Delta create conditions favoring reassortment, as evident from the gene constellations identified by this study. In conclusion, we present for the first time a comprehensive phylogenetic analysis of full genome sequences from low pathogenic avian influenza viruses circulating in Egypt, underscoring the significance of the region for viral reassortment and the potential emergence of novel avian influenza A viruses, as well as representing a highly diverse influenza A virus gene pool that merits continued monitoring.

Hemorrhagic fever with renal syndrome (HFRS) and Crimean-Congo hemorrhagic fever (CCHF) are the 2 widespread viral hemorrhagic fevers occurring in Europe. HFRS is distributed throughout Europe, and CCHF has been reported mainly on the Balkan Peninsula and Russia. Both hemorrhagic fevers are endemic in Bulgaria. We investigated to what extent acute undifferentiated febrile illness in Bulgaria could be due to hantaviruses or to CCHF virus. Using enzyme-linked immunosorbent assays (ELISAs), we tested serum samples from 527 patients with acute febrile illness for antibodies against hantaviruses and CCHF virus. Immunoglobulin M (IgM) antibodies against hantaviruses were detected in 15 (2.8%) of the patients. Of the 15 hantavirus-positive patients, 8 (1.5%) were positive for Dobrava virus (DOBV), 5 (0.9%) were positive for Puumala virus (PUUV), and the remaining 2 were positive for both hantaviruses. A plaque reduction neutralization test (PRNT) confirmed 4 of the 10 DOBV-positive samples. PRNT was negative for all PUUV-positive samples. Serologic evidence of recent CCHF virus infection was found in 13 (2.5%) of the patients. Interestingly, HFRS and CCHF were not only detected in well-known endemic areas of Bulgaria but also in nonendemic regions. Our results suggested that in endemic countries, CCHF and/or HFRS might appear as a nonspecific febrile illness in a certain proportion of patients. Physicians must be aware of possible viral hemorrhagic fever cases, even if hemorrhages or renal impairment are not manifested.

We analyzed highly pathogenic avian influenza A(H5N1) viruses isolated from humans infected in Egypt during 2007-2011. All analyzed viruses evolved from the lineage of subtype H5N1 viruses introduced into Egypt in 2006; we found minimal evidence of reassortment and no exotic introductions. The hemagglutinin genes of the viruses from 2011 formed a monophyletic group within clade 2.2.1 that also included human viruses from 2009 and 2010 and contemporary viruses from poultry; this finding is consistent with zoonotic transmission. Although molecular markers suggestive of decreased susceptibility to antiviral drugs were detected sporadically in the neuraminidase and matrix 2 proteins, functional neuraminidase inhibition assays did not identify resistant viruses. No other mutations suggesting a change in the threat to public health were detected in the viral proteomes. However, a comparison of representative subtype H5N1 viruses from 2011 with older subtype H5N1 viruses from Egypt revealed substantial antigenic drift.

To evaluate the effectiveness of an intensive hand hygiene campaign on reducing absenteeism caused by influenza-like illness (ILI), diarrhea, conjunctivitis, and laboratory-confirmed influenza, we conducted a randomized control trial in 60 elementary schools in Cairo, Egypt. Children in the intervention schools were required to wash hands twice each day, and health messages were provided through entertainment activities. Data were collected on student absenteeism and reasons for illness. School nurses collected nasal swabs from students with ILI, which were tested by using a qualitative diagnostic test for influenza A and B. Compared with results for the control group, in the intervention group, overall absences caused by ILI, diarrhea, conjunctivitis, and laboratory-confirmed influenza were reduced by 40%, 30%, 67%, and 50%, respectively (p<0.0001 for each illness). An intensive hand hygiene campaign was effective in reducing absenteeism caused by these illnesses.

Dengue fever (DF)/dengue hemorrhagic fever (DHF) has emerged as a global public health problem with countries in Asia and the Pacific sharing more than 70% of the disease burden. In 2004-2005 a total of 312 cases admitted to Pediatric and Sea Port Hospitals in Port Sudan were clinically diagnosed as DHF. The mortality rate recorded was 3.8% (n=12). Of the cases 73.4% were patients 5-15 years of age. A total of 91.2% of cases were admitted during May and June 2005 with 49.4% residing in the eastern region of Port Sudan. Dengue shock syndrome was observed in 37 of 312 (11.9%). All patients had thrombocytopenia with platelets count ranged from <100,000 to <150,000 cell/mm³. Of the 40 sera tested using RAPID-cassette test in the Khartoum Central Public Health Lab, 36 (90%) were dengue IgM positive. A subset of these sera (n=23) were sent to NAMRU-3 and confirmed by IgM-capture ELISA; 9 of 23 were PCR positive for dengue serotype 3., (Published by Elsevier Ltd.)

We conducted a prospective cohort study to determine prevalence and incidence of West Nile virus (WNV) in Egypt. Cohorts were established in Upper (UE), Middle (ME), and Lower (LE) Egypt. Additionally, a cross-sectional serosurvey was performed in the North (NS) and South (SS) Sinai. Cohorts were bled initially and 1 year later. Sera were tested for WNV-IgG by ELISA and positive sera were confirmed by plaque reduction neutralization test (PRNT). Sentinel chicken flocks placed in the above sites were bled monthly for virus isolation and serology. Mosquitoes were collected monthly from the above sites and tested for WNV. Human seroprevalence rates were 35%, 27%, 14%, 1% and 7% in UE, ME, LE, NS and SS, respectively. Seroconversion rates were 18%, 17% and 7% in UE, ME and LE, respectively; 49% of the seroconverters reported undiagnosed febrile illness. Sentinel chickens showed seroconversion in all study sites. WNV was isolated from both sentinel chickens and mosquitoes in cohort sites. This study demonstrates that WNV was actively circulating during the study period in different areas in Egypt and causing febrile illness in a considerable proportion of individuals in the study sites., (Published by Elsevier Ltd.)

In Egypt, the etiology of chronic renal failure (CRF) is not well defined. A hospital-based case-control study was initiated in February 1998, to determine whether hantavirus infection is involved in chronic renal disease (CRD) in Egypt. The study enrolled 350 study patients with a history of CRF and 695 matched controls with CRD due to renal calculus or renal cancer, but with normal renal functions. Sera from cases and controls were tested for anti-hantavirus IgG using ELISA with a cell-lysate antigen from Hantaan virus prototype strain 76-118. A demographic questionnaire was completed for each study participant. Five of the 350 cases (1.4%), and seven of the 695 controls (1.0%) were antibody-positive to hantavirus, with a titer > or =1:400. The difference in antibody prevalence between the study cases and the control cases was not statistically significant (P = 0.48). All antibody-positive study cases and controls had been exposed to rodents. Data indicated that in Egypt, hantavirus seroprevalence in CRD patients is low, and hantavirus infections do not appear to be a significant cause of CRF.

We conducted a serosurvey among patients of a health center in Hashimiah, a Jordanian town of 30,000 inhabitants located near a wastewater treatment plant and its effluent channel. Serum samples from 261 patients >/=5 years of age were assessed for immunoglobulin G (IgG) and IgM antibodies against West Nile, sandfly Sicilian, sandfly Naples, and Rift Valley viruses; the seroprevalence of IgG antibodies was 8%, 47%, 30%, and 0%, respectively. Female participants were more likely to have been infected than male. Persons living within 2 km of the treatment plant were more likely to have been infected with West Nile (p=0.016) and sandfly Sicilian (p=0.010) viruses. Raising domestic animals within the house was a risk factor for sandfly Sicilian (p=0.003) but not for sandfly Naples virus (p=0.148). All serum samples were negative for IgM antibodies against the tested viruses. Our study is the first documentation of West Nile and sandfly viruses in Jordan and calls attention to the possible health hazards of living close to wastewater treatment plants and their effluent channels.

The objective of the present work is to evaluate Fasciola E/S antigens for diagnosis of early fascioliasis utilizing different diagnostic techniques. Using enzyme-linked immunoelectro-transfer blot (EITB), Fasciola-specific E/ S protein fraction band (49.5 kDa) was determined and electroeluted. The mono-specific antibodies against this specific fraction band were prepared by immunizing pathogen-free rabbit. Assessment of the prepared mono-specific antibodies in diagnosis of human fascioliasis was performed through the detection of E/S copro-antigens by enzyme-linked immunosorbent assay (ELISA) in stool eluates obtained from patients with confirmed fascioliasis, other parasites as well as from other healthy individuals. Serum samples were collected and tested to detect serum antibodies against Fasciola E/S antigen using EITB and counter immunoelectrophoresis (CIEP). Analysis of Fasciola adult worm E/S products by SDS/PAGE revealed a number of bands, the molecular weight (MW) of which ranged from 14-200 kDa; with three major bands (27.5, 32.5 and 55 kDa). Fasciola EIS 49.5 kDa protein fraction proved to be specific to F. gigantica. Cross reaction with S. mansoni was observed at higher MW (110-120 kDa). The sensitivity, specificity and diagnostic accuracy of EITB were 45.2%, 100% and 70.7%, respectively, while those of CIEP were 38.7%, 100% and 67.2%, respectively. ELISA technique using mono-specific anti-49.5 kDa to detect copro-antigens proved to be practical and reliable. It showed higher sensitivity (91.4%) and higher diagnostic accuracy (91.8%), while the specificity was 92.3%. In addition, ELISA had higher negative predictive value (88.9%) and fair positive predictive value (94.1%).

Rabbit monospecific antibody Leishmania major surface glycoprotein (gp63) was used in an indirect immunofluorescence assay (IFA) to identify promastigotes and amastigotes from sandflies and lesion fluid aspirates, respectively. Anti-gp63 fluoresced the entire periphery and flagella of promastigotes of different Leishmania species at dilutions to 1:50. Promastigotes were distinctively demonstrated in whole infected sandfly homogenate for 17 days post-infection. Amastigotes from lesion aspirates of infected BALB/c mice and from a human cutaneous leishmaniasis case were also fluoresced by the antibody. This technique could prove to be especially useful in arthropod vector surveillance efforts in which detection of the pathogen is as important as speciation of the sand fly vector.

Stool and blood specimens were collected from each of 404 and 576 individuals at Sindbis village (Qualiubia Governorate) in the Nile Delta where schistosomiasis is endemic and El-Rashda village (New Valley Governorate) in the Western Desert of Egypt where there is no schistosomiasis; respectively. Based on the microscopical examination of stool specimens, the prevalence of infection with Entamoeba (E. histolytica and/or E. dispar which are morphologically indistinguishable) was higher at Sindbis than at El Rashda village (29.3% and 20%, respectively). At Sindbis, the prevalence of Entamoeba (both species) was 35.2% (50/142) in S. mansoni infected individuals versus 26.3% (69/262) in S. mansoni negative individuals. Serum antibodies develop only against E. histolytica but not against E. dispar infection. When serological results were considered, the prevalence of E. histolytica was 4.7% in Sindbis and 3.4% at El Rashda based on those who were positive microscopically and serologically in the two villages, respectively. In other words, only 16-17% of those who were positive microscopically can be considered infected with E. histolytica as determined serologically. However, the prevalence of E. histolytica (present or past) based on those who were positive serologically whether positive or negative microscopically was 13.4% and 12.7% at the two villages, respectively. At Sindbis, the prevalence of E. histolytica infection was lower in S. mansoni negative (8.5%) than in S. mansoni positive (16.0%) individuals. These epidemiologic data suggest that: (1) S. mansoni infection may suppress the immune response of the host and therefore, the prevalence of E. histolytica based on serological testing is probably underestimated in the S. mansoni infected people and it may be higher than in the S. mansoni negative people. (2) Serological examinations can be used in determining the true prevalence of E. histolytica (present or past infections) until a routine test for detecting E. histolytica specific antigen in stool becomes available to differentiate E. histolytica from E. dispar infections.

The life cycle of Leishmania consists of two distinct developmental stages: the amastigote, which is the ovoid non-flagellated form found in the vertebrate host, and the promastigote, which is an elongated flagellated form found in the gut of an infected sandfly. Following its injection into the vertebrate host by the sandfly vector, the promastigote transforms into an amastigote after entering the host macrophage. The environmental cues inducing this transformation are not fully understood. Attempts to axenically develop and cultivate amastigotes from different Leishmania species have indicated that species and sometimes even strains of the same species vary in their requirements for this process (Pan et al., 1993). The majority of the available published data on transformation pertains to New World Leishmania and provides evidence that elevation in the incubation temperature and/or acidic pH can in some cases induce amastigote formation from promastigotes.

Promastigotes of a viscerotropic Leishmania tropica strain, originally isolated from a U.S. veteran returning from the Gulf War, was studied for extracellular transformation to amastigotes. Promastigotes were grown in three different media namely, Schneider's Drosophila, Dulbecco's modified Eagle Medium (D-MEM) and RPMI-1640. For each medium the effect of low pH (5.8), elevated temperature (34 degrees C) and the effect of both conditions combined were compared. Cells were counted and examined for morphological changes daily. Transformation to morphologically typical amastigotes occurred in D-MEM and Schneider's media at each of the three different experimental conditions with maximum transformation occurring on day eight in D-MEM maintained at pH 5.8 and 34 degrees C. Complete transformation was not achieved in RPMI-1640 under any of the experimental conditions described. Transformed amastigotes could successfully infect murine macrophages indicating biological competence. Amastigotes could not be propagated under any of the conditions described herein.

Giardia lamblia is recognized as one of the most common agents for diarrhea world wide. To date, microscopical examination of stool samples is the gold standard for giardiasis diagnosis. However, intermittence of the Giardia cycle and some medications may cause temporary disappearance of cysts from stools, thus giving false negative results. In the present study, we evaluated a commercially available enzyme immunoassay kit (GiardEIA) for the detection of Giardia copro-antigens and compared the results with those of the merthiolate-iodine-formaldehyde concentration (MIFC) microscopical examination technique. Sixty-nine fecal samples from children 2-12 years old were emulsified and allowed to react with a Giardia specific antibody, then with an enzyme conjugated antibody and the reaction was developed colorimetrically. Seventy-four percent of the parasitologically positive Giardia cases were also positive by GiardEIA while 26% of the microscopically negative cases were positive by the assay. GiardEIA gave negative results with 82% and 100% of stools with helminthic and protozoan (other than Giardia) infections, respectively. Similarly, no cross-reactivity was found with any of the bacterial agents including Shigella flexneri, pathogenic E. coli, Klebsiella spp. and Salmonella typhi. GiardEIA is a simple assay that can diagnose 24 samples in less than an hour without the need for any special equipment and can be useful in epidemiological surveys and in giardiasis outbreaks.

During the period from October 1982 to July 1985 cutaneous leishmaniasis was diagnosed in 113 Multinational Force and Observers (MFO) soldiers from various nations, East Sinai. Leishmania was isolated from twelve of these patients and maintained in vitro using Tanabe's medium. Intrasplenic inoculation of promastigotes from isolate cultures into hamsters resulted in dissemination patterns typical of cutaneous leishmaniasis. The excreted factor in culture when serotyped against known marker strains indicated subserotype A1B2. They also exhibited similar banding patterns to L. major L-137 for GPI, G 6-PD, MDH, NH, MPI, PGM and variant B for 6-PGDH. This leishmaniasis focus appears epidemiologically similar to that of nearby Mid Western Negev where transmission involves Phlebotomus papatasi, Psammomys, and Meriones.

Two visceral Leishmania isolates from children (aged 1 1/2 and 4 years) living in El Agamy area, Alexandria, Egypt, were compared with 5 marker strains, and 2 other human isolates from Sinai and Sudan, identified on clinical and geographical grounds as cutaneous and visceral leishmaniasis respectively. Isoenzyme variations were assessed on the basis of their electrophoretic profiles on cellulose acetate membranes. The enzymes studied were glucose 6-phosphate dehydrogenase E.C.1.1.1.49, phosphoglucomutase E.C.2.7.5.1, 6-phosphogluconate dehydrogenase E.C.1.1.1.44 (6-PGD), glucose phosphate isomerase E.C.5.3.1.9, malate dehydrogenase E.C.1.1.1.37, mannose phosphate isomerase E.C.5.3.1.8 and nucleoside hydrolase E.C.3.2.2.2. The last 4 enzymes could differentiate between cutaneous and visceral strains. The Alexandria strains proved to belong to the L. donovani complex; however, their 6-PGD pattern was identical to that of L. infantum, which was different from that of the L. donovani marker strain.

Disease concepts and medical treatment practices surrounding schistosomiasis haematobium were studied among males in Upper Egyptian villages and towns using interview methods. Most informants considered bilharzia to be a serious disease for which they commonly sought treatment. Its occurrence was attributed primarily to natural causes, particularly various aquatic worms and insects, dirts, excrement, dead animals, toxins and stagnant and vegetated waters, mostly large canals. Contact with water from the Nile river was generally thought to be quite safe. Drug treatment was weakly associated with amount of education. All groups reported use of antischistosomal drugs and plant medicines. Seventy-four per cent of the sample had a treatment history, 64% having taken oral drugs and/or injections, 40% plant medicines and 29% both. Drinking decoctions of damsissa (Ambrosia maritima) was the most commonly used household remedy. Plant materials were usually obtained from fields, gardens and local markets and patent medicines from nearby clinics and private physicians in towns. Recommendations are made for the national mass chemotherapy programme.

1. The pattern and activity of isocitrate, lactate and malate dehydrogenases and malic enzyme were studied in plasma of normal hamsters and hamsters at the 26th day of infection with S. mansoni. 2. Although the electrophoretic patterns of these enzymes were similar in normal and infected hamsters, their activities were higher in the latter than the former group of animals. The elevation in the enzymic activity indicates that there is tissue damage caused by the larvae at this stage.