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Background: The female lower genital tract is a gateway for pathogens entering the host through the mucous membrane. One of the prevalent human viruses is Torque teno virus (TTV). The major reported routes of TTV transmission are fecal-oral and parenteral. Furthermore, other modes of transmission, e.g. sexual contact, are suggested. To investigate the sexual route of TTV transmission, cervical smears of healthy women and those with cervical lesions were screened for the presence of TTV DNA., Methods: TTV DNA was studied in cervical smears of 95 patients with cervical lesions and 55 healthy women. Paired serum samples were available from 55 and 42 women, respectively. All healthy women had normal cytology while 44 patients had histologically confirmed low-grade lesion (LGL) and 51 high-grade lesion (HGL). TTV DNA was detected with primers specific for the non-coding region. In 40 paired cervical smears and serum samples, the phylogenetic group of TTV isolates was determined. The presence of HPV DNA in cervical smears was detected by means of PCR with MY09/11 primers., Results: The prevalence of TTV DNA in cervical smears of healthy women was 52.7% and was comparable with that in paired serum samples (50%). Symptomatic women had significantly higher prevalence of TTV DNA in cervical smears (74.7%) than healthy controls. The TTV DNA prevalence in patient serum samples was 51%. The phylogenetic groups of TTV serum isolates were concordant with those of TTV from cervical smears of the same subjects. In cervical smears, a wider variety of TTV isolates was found. The viral loads in cervical smears were 10 to 1000 times as high as in sera. The HPV-positive study subjects had significantly higher TTV DNA prevalence than HPV negatives. The prevalence of TTV was not associated with disease severity., Conclusion: High prevalence of TTV in cervical smears suggests that sexual transmission is another mode of expansion of TTV infection among the population. The higher viral load in cervical smears than in the respective serum samples might indicate active TTV replication in the female genital tract. Nevertheless, cooperation between TTV and HPV needs to be further investigated.

Sera and selected tissue homogenates collected from gnotobiotic swine never exposed to the environment or other swine tissues were tested for the presence of porcine torque teno virus (TTV) DNAs by nested and non-nested polymerase chain reactions (PCR) using primers specific for the untranslated region of porcine genogroups (g) 1 and 2. Twenty-three of 105 (21.9%) gnotobiotic piglets were g1- and/or g2-TTV DNA positive. Twenty-three of 27 (85.2%) sow sera, collected at the time of Caesarian derivation of the litters contained either or both TTV genogroup DNAs. These data demonstrate that porcine TTV may be transmitted to piglets by the in utero route and that the incidence of fetal infection is high.

Routes of swine torque teno virus (TTV) transmission have been minimally investigated in the pig population. Current knowledge suggests the faecal-oral route as the most probable way of viral dissemination. Other transmission routes, such as mother-to-infant, have been studied in humans, but no information is available for swine. Thus, the objective of the present study was to determine the prevalence of two swine TTV genogroups, TTV1 and TTV2, in colostrum samples (n=61) and sera samples from sows (n=10) and stillborn pigs coming from them (n=30). Colostrum was fractioned into two components, milk whey and cell pellets, and 26 out of 61 milk whey samples and 30 out of 58 cell pellets analyzed contained TTV1 or TTV2 genomes, respectively, detected by specific PCR methods. Six and 3 out of 10 serum samples from sows were positive for TTV1 and TTV2 DNA, respectively. Finally, 15 out of 30 sera from stillborns were PCR positive for TTV1, but only 2 were TTV2 positive. Positive stillborns were always infected with the same TTV genogroup as their mothers. However, TTV sequence analysis determined that sequences obtained from sows and their stillborns were not identical. In conclusion, our results indicated that swine TTVs can be transmitted vertically, and suggest that different sow-to-piglet transmission routes may coexist, including transplacental/intra-uterine as well as through lactation. This study represents the first description of swine TTV presence in colostrum and stillborn piglets.

Humans are frequently infected with three anelloviruses which have circular DNA genomes of 3.6-3.9 kb [Torque teno virus (TTV)], 2.8-2.9 kb [Torque teno mini virus (TTMV)] and 3.2 kb [a recently discovered anellovirus named Torque teno midi virus (TTMDV)]. Unexpectedly, human TTMDV DNA was not detectable in any of 74 chimpanzees tested, although all but one tested positive for both human TTV and TTMV DNA. Using universal primers for anelloviruses, novel variants of TTMDV that are phylogenetically clearly separate from human TTMDV were identified from chimpanzees, and over the entire genome, three chimpanzee TTMDV variants differed by 17.9-20.3 % from each other and by 40.4-43.6 % from all 18 reported human TTMDVs. A newly developed PCR assay that uses chimpanzee TTMDV-specific primers revealed the high prevalence of chimpanzee TTMDV in chimpanzees (63/74, 85 %) but low prevalence in humans (1/100). While variants of TTV and TTMV from chimpanzees and humans were phylogenetically interspersed, those of TTMDV were monophyletic for each species, with sequence diversity of <33 and <20 % within the 18 human and three chimpanzee TTMDV variants, respectively. Maximum within-group divergence values for TTV and TTMV were 51 and 57 %, respectively; both of these values were substantially greater than the maximum divergence among TTMDV variants (44 %), consistent with a later evolutionary emergence of TTMDV. However, substantiation of this hypothesis will require further analysis of genetic diversity using an expanded dataset of TTMDV variants in humans and chimpanzees. Similarly, the underlying mechanism of observed infrequent cross-species infection of TTMDV between humans and chimpanzees deserves further analysis.

The commercial trade of wildlife occurs on a global scale. In addition to removing animals from their native populations, this trade may lead to the release and subsequent introduction of nonindigenous species and the pathogens they carry. Emerging infectious diseases, such as chytridiomycosis caused by the chytrid fungus Batrachochytrium dendrobatidis (Bd), and ranaviral disease have spread with global trade in amphibians and are linked to amphibian declines and die-offs worldwide, which suggests that the commercial trade in amphibians may be a source of pathogen pollution. We screened tiger salamanders involved in the bait trade in the western United States for both ranaviruses and Bd with polymerase chain reaction and used oral reports from bait shops and ranavirus DNA sequences from infected bait salamanders to determine how these animals and their pathogens are moved geographically by commerce. In addition, we conducted 2 surveys of anglers to determine how often tiger salamanders are used as bait and how often they are released into fishing waters by anglers, and organized bait-shop surveys to determine whether tiger salamanders are released back into the wild after being housed in bait shops. Ranaviruses were detected in the tiger salamander bait trade in Arizona, Colorado, and New Mexico, and Bd was detected in Arizona bait shops. Ranaviruses were spread geographically through the bait trade. All tiger salamanders in the bait trade were collected from the wild, and in general they moved east to west and north to south, bringing with them their multiple ranavirus strains. Finally, 26-73% of anglers used tiger salamanders as fishing bait, 26-67% of anglers released tiger salamanders bought as bait into fishing waters, and 4% of bait shops released tiger salamanders back into the wild after they were housed in shops with infected animals. The tiger salamander bait trade in the western United States is a useful model for understanding the consequences of the unregulated anthropogenic movement of amphibians and their pathogens through trade.

Objective: To determine whether commercial Mycoplasma hyopneumoniae bacterins sold for use in swine contain porcine torque teno virus (TTV)., Sample Population: 22 commercially available M hyopneumoniae bacterins., Procedures: Direct and nested PCR assays for genogroup-specific TTV DNAs were performed on serials of M hyopneumoniae bacterins by use of published and custom-designed primer pairs at 3 laboratories in North America and Europe., Results: Of the 22 bacterins tested by use of direct and nested PCR assays, 7 of 9 from the United States, 2 of 5 from Canada, and 4 of 8 from Europe contained genogroup 1- and genogroup 2-TTV DNAs. In some bacterins, the TTV DNAs were readily detected by use of direct PCR assays., Conclusions and Clinical Relevance: Analysis of these data indicated that many of the commercially available M hyopneumoniae bacterins were contaminated with TTV DNA. It is possible that some of these bacterins could inadvertently transmit porcine TTV infection to TTV-naïve swine.

A diverse class of proteins called lectins plays a major role in shrimp innate immunity. In this study, the cDNA encoding a C-type lectin of Penaeus monodon (PmLT) was cloned, and its potential role examined. Despite the low overall amino acid sequence identity with other animal lectins, PmLT includes conserved carbohydrate recognition domains (CRDs) characteristic of animal C-type lectins. Unlike the other two P. monodon lectin-like proteins described to date that have one CRD, PmLT has two CRDs. The first CRD contains a QPD motif with specificity for binding galactose, while the second CRD contains a EPN motif for binding mannose. PmLT transcripts can be detected in the hepatopancreas but not in other tissues. Expression studies showed that PmLT mRNA transcript level decreased initially and then gradually increased after whole shrimp or hepatopancreas tissue fragments were treated with white spot syndrome virus (WSSV) extract but were not affected by bacteria. Using anti-rPmLT antibody, PmLT was detected only in the hepatopancreas specific F cells (Hpf). In vitro encapsulation assay showed that agarose beads coated with rPmLT were encapsulated by hemocytes indicating a role in innate immune response. In summary, PmLT is produced in the hepatopancreas and may act as a pattern recognition protein for viral pathogens and also activates the innate immune responses of the shrimp to bacteria. The dual-CRD structure of PmLT may assist the recognition of diverse pathogens.

Current clinical studies on human annelloviruses infections are directed towards finding an associated disease. In this review we have emphasized the many similarities between human anellovirus and avian circoviruses and the cell and tissue types infected by these pathogens. We have done this in order to explore whether knowledge acquired from natural and experimental avian infections could reflect and be extrapolated to the less well-characterized human annellovirus infections. The knowledge gained from the avian system may provide suggestions for decoding the enigmatic human anellovirus infections, and finding the specific disease or diseases caused by these human anellovirus infections. Each additional parallelism between chicken anemia virus (CAV) and Torque teno virus (TTV) further strengthens this premise. As we have seen information from human infections can also be used to better understand avian infections as well. Increased attention must be focused on the "hidden" or unrecognized, seemingly asymptomatic effects of circovirus and anellovirus infections. Understanding the facilitating effect of these infections on disease progression caused by other pathogens may help to explain differences in outcome of complicated poultry and human diseases. The final course of a pathogenic infection is determined by variations in the state of health of the host before, during and after contact with a pathogen, in addition to the phenotype of the pathogen and host. The health burden of circoviridae and anellovirus infections may be underestimated, due to lack of awareness of the need to search past the predominant clinical effect of identified pathogens and look for modulation of cellular-based immunity caused by co-infecting circoviruses, and by analogy, human anneloviruses.

A combined functional investigation on the four per os infectivity factors (PIFs) of Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus (HearNPV) was conducted in this study. HearNPV bacmids with deletions of p74 (Ha20), pif1 (Ha111), pif2 (Ha132) and pif3 (Ha98) were constructed individually by homologous recombination in Escherichia coli cells. Repaired bacmids with respective pifs were also constructed. Western blot analyses revealed that all four PIFs were structural components of the envelope of HearNPV occlusion-derived virus (ODV). Electron microscopy showed that deletion of the pifs did not have any obvious effects on the morphology of the occlusion bodies (OBs). Bioassay analyses indicated that deletion of any of the above pifs resulted in loss of oral infectivity of OBs. The mixtures of the four pif-deletion mutants also resulted in deficiency of oral infectivity, implying that the four PIFs must be structural components of the same ODV to accomplish their function. Repairing of the respective genes into the pif-deletion bacmids could rescue the oral infectivity of the pif-deletion viruses. Calcofluor, which can damage the peritrophic membrane (PM), could not rescue the defects of the oral infectivity of the pif-deletion viruses, indicating that the PM is not likely to be the functional target of the PIFs.

1. The hypothesis that habitat fragmentation (biotic or abiotic) alters the transmission of disease within a population is explored using field data from a well-studied amphibian-pathogen system. 2. We used the Ambystoma tigrinum-A. tigrinum virus (ATV) model system to show how habitat fragmentation as a result of emergent vegetation and habitat management affects disease transmission dynamics in ponds across a landscape. 3. We quantified variation in ATV infection over time and across the landscape. ATV infection was significantly higher in ponds modified for livestock use (P = 0.032). Disease incidence decreased with increased amounts of emergent vegetation (P < 0.001). These factors appear to control disease transmission by altering the host contact rate and with it disease transmission. 4. A field experiment to test the effect of emergent vegetation on the distribution of larvae in ponds demonstrated a behavioural change in larvae found in sparsely vegetated ponds. Microhabitat choices resulted in larvae being concentrated at the pond edge resulting in a 'halo effect' in sparsely vegetated ponds, whereas larvae in heavily vegetated ponds were distributed more evenly throughout. Microhabitat choice affects the effective density that larvae experience. This 'halo effect' increases contact rates in the shallows of sparsely vegetated ponds and increases the transmission of a directly transmitted pathogen. 5. Despite recurrent epidemics of a lethal Ranavirus in tiger salamanders on the Kaibab Plateau, Arizona, USA, these populations persist. We discuss the implications of our results in the context of density-dependent transmission and homogeneous mixing, two increases key assumptions of epidemiological theory.

Frog virus 3 (FV3) and FV3-like viruses, are members of the genus Ranavirus (family Iridoviridae), and they have been associated with infectious diseases that may be contributing to amphibian population declines. We examined the mode of transmission of an FV3-like virus, and potential hosts and reservoirs of the virus in a local amphibian community. Using the polymerase chain reaction to detect infected animals, we found an FV3-like virus in south-central Ontario, Canada, amphibian communities, where it infects sympatric amphibian species, including ranid and hylid tadpoles (Rana sylvatica, Hyla versicolor, and Pseudacris spp.), larval salamanders (Ambystoma spp.), and adult eastern-spotted newts (Notophthalmus viridescens). The high prevalence of FV3-like infections in caudate larvae suggests that salamanders are likely to be both hosts and reservoirs. In laboratory FV3 challenges of R. sylvatica, the rate of infection was dependent on the amount of virus to which the animals were exposed. In addition, although vertical transmission was suspected, horizontal transmission through exposure to infected pond water is the most likely route of infection in tadpoles. Based on our observations, a simple model of FV3/FV3-like virus transmission postulates that, in aquatic amphibian communities, transmission of the virus occurs between anuran and urodele species, with ambystomatid salamanders the most likely reservoir for the ranavirus in our study.

Distinguishing whether pathogens are novel or endemic is critical for controlling emerging infectious diseases, an increasing threat to wildlife and human health. To test the endemic vs. novel pathogen hypothesis, we present a unique analysis of intraspecific host-pathogen phylogenetic concordance of tiger salamanders and an emerging Ranavirus throughout Western North America. There is significant non-concordance of host and virus gene trees, suggesting pathogen novelty. However, non-concordance has likely resulted from virus introductions by human movement of infected salamanders. When human-associated viral introductions are excluded, host and virus gene trees are identical, strongly supporting coevolution and endemism. A laboratory experiment showed an introduced virus strain is significantly more virulent than endemic strains, likely due to artificial selection for high virulence. Thus, our analysis of intraspecific phylogenetic concordance revealed that human introduction of viruses is the mechanism underlying tree non-concordance and possibly disease emergence via artificial selection.

Frog virus 3 (FV3) or FV3-like viruses (Iridoviridae) infect a wide range of amphibian species, and they are becoming increasingly and causally associated with amphibian disease outbreaks worldwide. We have established the frog Xenopus laevis as an experimental model to study host defense and pathogenesis of FV3 infection. Although X. laevis adults usually clear FV3 infection within a few weeks, viral DNA has been detected in the kidneys several months after they had been experimentally infected; virus also has been detected in seemingly healthy nonexperimentally infected adults. Based on this information, we hypothesized that covert FV3 infection may occur in Xenopus. We first conducted a survey that detected FV3 by polymerase chain reaction (PCR) in the kidneys (the main site of FV3 infection) in a significant fraction of X. laevis raised in five different locations in the United States. Asymptomatic FV3 carriers also were detected by initiation of an acute systemic FV3 infection in frogs that had been immunosupressed by sublethal gamma-irradiation. Finally, we focused on macrophages as a potential site for viral persistence, and we showed that FV3 can infect peritoneal macrophages in vitro as determined by reverse transcriptase-PCR detection of viral mRNAs. Unlike kidney cell lines that are readily killed by FV3, infected macrophages, like uninfected macrophages, survived up to 12 days. Viral transcription also was detected in macrophages from animals up to 12 days after infection. These results suggest that FV3 can become quiescent in resistant species such as Xenopus, thereby making these species potential viral reservoirs.

During investigations of epidemic frog mortality in Britain, a novel fatal systemic haemorrhagic disease of common toads was discovered. This disease resembles a systemic haemorrhagic disease of common frogs in Britain, which is one of a range of fatal disease syndromes, characterized by systemic haemorrhages, skin ulceration or a combination of these lesions, caused by ranavirus infection. Ranavirus previously isolated from diseased toads was inoculated into common frogs to evaluate if this virus could infect and cause disease in common frogs. All virus-inoculated frogs died with systemic haemorrhages between 6 and 8 days post-inoculation, giving similar results to those produced by the inoculation of frogs with ranavirus cultured from naturally diseased frogs. These results indicate that the same, or similar, viruses are affecting both frogs and toads in the field and confirm that ranavirus has emerged as an important cause of amphibian mortality in Britain.

Transmission is central to pathogen fitness and strongly influences the impact of pathogens on host populations. Particularly important to transmission dynamics is the distinction between direct transmission requiring close physical contact (e.g. bumping, fighting, or coughing) and indirect transmission from environmental sources such as contaminated substrates. We present data from 4 experiments addressing the form, routes, and timing of transmission of Ambystoma tigrinum virus (ATV) among tiger salamanders Ambystoma tigrinum nebulosum. Our data suggest that ATV is efficiently transmitted by direct interactions between live animals (bumping, biting and cannibalism) as well as by necrophagy and indirectly via water and fomites. Determining which form of transmission is most important in nature is essential for understanding transmission at the population level. Our experiments also revealed an important temporal aspect of infectiousness: larval salamanders become infectious soon after exposure to ATV and their propensity to infect others increases with time. These results begin to clarify the mechanisms and dynamics of ATV transmission and lead to key questions that need to be addressed in future research.

An experimental transmission study was designed to determine whether a causal relationship exists between a Ranavirus (BSTRV) isolated from a Burmese star tortoise that died and the lesions observed in that tortoise. A pilot study was performed with 3 box turtles (Terrapene ornata ornata) and 3 red-eared sliders (RESs; Trachemys scripta elegans) to assess their suitability in a larger study. Based on the outcome of this study, RESs were selected, and 2 groups of 4 RESs received either an oral (PO) or intramuscular (IM) inoculum containing10(5) 50% Tissue Culture Infecting Dose (TCID(50)) of a BSTRV-infected cell lysate. One turtle each was mock inoculated PO or IM with the same volume of uninfected cell lysate. Three of four IM-inoculated RESs developed clinical signs (nasal and ocular discharge [3 of 3], oral plaques [1 of 3], conjunctivitis and hyphema [1 of 3] and extreme lethargy [3 of 3]). A Ranavirus was isolated from kidney homogenates of 3 euthanatized turtles; DNA sequences of a portion of the major capsid protein gene were amplified by polymerase chain reaction. Consistent histologic lesions were observed only in IM-inoculated turtles and included fibrinoid vasculitis centered on splenic ellipsoids, multifocal hepatic necrosis, and multicentric fibrin thrombi in a variety of locations, including hepatic sinusoids, glomerular capillary loops, and pulmonary capillaries. Virions compatible with Ranavirus were observed within necrotic cells of the spleen of 1 IM-inoculated turtle using transmission electron microscopy. This study fulfills Koch's postulates, confirming a causal relationship between BSTRV and the clinical and histologic changes in chelonians infected with this virus.

SEN is a newly discovered blood-transmissible virus. Among its variants, SENV-D and -H are most often associated with non-A, -E hepatitis. Very little is known about the risk of vertical transmission of the virus. By using polymerase chain reaction with specific primers for SENV-D and -H, we investigated the prevalence of SENV-H and -D infection, the transmission rate of SENV infection and clinical features of SENV-infected children in 89 hepatitis C virus (HCV)-positive human immunodeficiency virus type 1-negative mothers. SENV infection was found in 36 (40%) mothers, and SENV-D was more frequent than SENV-H infection (34/36, 94%vs 5/36, 14%, P < 0.01). No difference in SENV infection rates was found between injection drug user (IDU) mothers (17/51, 33%) and mothers with no risk for bloodborne infection (19/38, 50%, P = ns). SENV-H infection was found only in IDU mothers and mothers with HCV genotype1b. Both SENV-D and -H can be transmitted to the offspring with an overall rate of 47%. Vertical transmission of HCV does not facilitate SENV infection of the offspring. Among 17 SENV-infected children, none was co-infected with HCV. Maternal HCV genotype or viral load does not interfere with mother-to-infant transmission of SENV. Persistence of SENV infection was demonstrated in 100% of infected children after 1-year follow-up, but none had clinical evidence of liver disease.

TT virus (TTV) is widespread throughout the world and can be detected in 50-95% of healthy individuals. However, in chronic hepatitis C virus (HCV) infected patients coinfected with TTV, histological activity indices were higher than patients with HCV infection alone. There are studies which indicate that TTV can cause aplastic anemia and thrombocytopenia. While TTV is known to be transmitted through blood transfusions and by fecal-oral route, published information on transplasental transmission is controversial. The aims of this study were to detect the frequency of TTV infection among healthy pregnant women and to search whether TTV is transmitted transplasentally during pregnancy. For this purpose, plasma samples collected from 54 women and their newborns were investigated by polymerase chain reaction (PCR), and viral loads were determined for infected newborns, and their respective mothers. Also, ten mothers whose newborns were tested negative for TTV-DNA were enrolled in viral load tests for comparison. TTV-DNA was detected in 49 (90.7%) of 54 women. Of 49 newborns whose mothers were infected with TTV, only 4 (8.2%) were found positive for TTV-DNA. There was no statistically significant difference in TTV-DNA loads between mothers who transmitted the virus and those who did not (Mann-Whitney U analysis Z=-0.071, P=0.944). As a result, transplasental transmission of TTV is possible but occurs in low frequency and independent of viral load.

The present review gives an updated overview of transfusion transmitted virus (TTV), a novel agent, in relation to its molecular characteristics, epidemiological features, modes of transmission, tissue tropism, pathogenesis, role in various diseases and its eradication from the body. TTV, a DNA virus, is a single stranded, non-enveloped, 3.8 kb long DNA virus with a small and covalently closed circular genome comprising 3852 bases. It was tentatively designated Circinoviridae virus. TTV genome sequence is heterogeneous and reveals the existence of six different genotypes and several subtypes. TTV has been reported to transmit not only via parenteral routes, but also via alternate routes. This virus has been detected in different non-human primates as well. At present, TTV is detected by polymerase chain reaction (PCR) with no other available diagnostic assays. It shows its presence globally and was detected in high percent populations of healthy persons as well as in various disease groups. Initially it was supposed to have strong association with liver disease; however, there is little evidence to show its liver tropism and contribution in causing liver diseases. It shows high prevalence in hemodialysis patients, pointing towards its significance in renal diseases. In addition, TTV is associated with several infectious and non-infectious diseases. Though, its exact pathogenesis is not yet clear, TTV virus possibly resides and multiplies in bone marrow cells and peripheral blood mononuclear cells (PBMCs). Recently, attempts have been made to eradicate this virus with interferon treatment. More information is still needed to extricate various mysteries related to TTV.

It is thought that white sturgeon iridovirus (WSIV) is transmitted vertically from adult white sturgeon Acipenser transmontanus to progeny, and that wild adults are carriers of this virus. Based on this assumption, egg disinfection trials were initiated using wild Kootenai River white sturgeon. Over 2 consecutive years, post-fertilized eggs were disinfected with iodine at concentrations ranging from 0 to 400 ppm. Eggs were incubated and progeny were reared on either de-chlorinated municipal or Kootenai River water. Juvenile sturgeon (mean weight 3.0 g) from these treatment groups were then subjected to a density stress (15 or 20 g(-1)) to manifest WSIV disease in individuals harboring the virus. In Year 1, mortality in all groups ranged from 6 to 37% and the use of municipal water was shown to significantly improve survival. However, WSIV infection was not detected in fish from any of the treatment groups or controls, and therefore did not contribute to the observed mortality. In Year 2, all treatment and control groups reared on Kootenai River water tested positive for WSIV infection and exhibited mortality ranging from 59 to 94%, but fish from groups reared on municipal water did not test positive for WSIV infection. This shows that that vertical transmission did not occur in this study. Horizontal transmission played a significant role in WSIV infection, but the lack of infection in Year 1 suggests a cyclic occurrence of the virus in the Kootenai River system. Although survival tended to be better in iodine-treated groups, the effects of iodine treatment in relation to WSIV transmission remain unknown. An important finding is that not all wild white sturgeon broodstock yield WSIV-positive progeny.

Background: SEN virus (SENV) is a newly discovered DNA virus. We conducted this study to evaluate potential modes of SENV transmission and the pathogenic effect of SENV on liver diseases in children., Methods: Polymerase chain reaction was used to detect 2 SENV variant (SENV-D and SENV-H) DNA in sera from healthy individuals and diseased children. Nucleotide sequence of SENV was determined by direct sequencing., Results: SENV infection was assessed in healthy individuals, including 50 newborns (sera collected from the umbilical cord), 24 infants, 46 preschool children (aged 1-6 years), 42 school children of an age before that of the first sexual experience (aged 7-12 years), 62 adolescents (13-18 years), 72 young adults (19-30 years) and 32 adults (>30 years). The prevalence of SENV-D and/or SENV-H (SENV-D/H) viremia in each group was 0%, 17%, 24%, 24%, 27%, 33% and 40%, respectively. The prevalence of SENV-D/H viremia in 18 children with non-A to E hepatitis, 64 thalassemic children, 80 children transfused during cardiac surgery, 30 children with chronic hepatitis B, 9 children with chronic hepatitis C and 32 infants with biliary atresia was 11%, 61%, 80%, 83%, 67% and 50%, respectively. SENV was found more frequently in all patient groups than in 174 age-matched controls (P < 0.01), with the exception of non-A to E hepatitis (11% versus 24% in the control group; P = 0.27). In 2 infants with proven intrauterine hepatitis B viral infection, identical SENV-D nucleotide sequence existed in both the maternal and neonate serum. Elevated alanine aminotransferase concentrations were rarely observed in children who acquired isolated SENV viremia because of transfusion for surgery. Infection with SENV in children with chronic hepatitis C virus or hepatitis B viral infection was not associated with higher peak alanine aminotransferase values., Conclusion: SENV is transmitted mainly via nonparenteral daily contact and frequently occurs early in life. Transfusion can significantly increase the rate of SENV viremia. SENV does not appear to cause hepatitis in children.

Members of the genus Ranavirus (family Iridoviridae) can cause catastrophic mortality of pond-breeding amphibians and are associated with an emerging infectious disease that may be contributing to amphibian declines. We conducted three experiments to examine factors that may affect transmission both within and between local breeding populations of the wood frog (Rana sylvatica). In a laboratory study, when exposed to moribund tadpoles collected during a local ranaviral die-off, uninfected tadpoles died as soon as 4 days after exposure. The onset of death was accelerated when tadpoles were allowed to scavenge on carcasses of infected tadpoles. In a mesocosm experiment that was conducted in outdoor wading pools, die-offs of tadpoles began approximately 19 days after infected tadpoles were added to pools containing uninfected tadpoles. Mass die-offs with greater than 98% mortality occurred in all pools, regardless of the initial tadpole density. In a second mesocosm experiment, the addition of water and bottom sediments that were collected from a pond during a ranaviral die-off did not result in lower tadpole survival or growth relative to controls. Only a small percentage of tadpoles appeared to be sick, and most tadpoles survived until the first individuals began metamorphosing within a pool. However, tests for ranavirus using polymerase chain reaction were positive for most pools that received contaminated sediment, suggesting that some infections were sublethal. Our results indicate that transmission within ponds is enhanced by scavenging and that spread between local ponds could occur via the transport of contaminated sediment by animals or humans.

The aim of the study was to assess the prevalence, clinical impact and importance of different risk factors of transmission of TTV infection in Slovakia using two PCR methods. Sera of 426 adult persons were examined. TTV DNA was identified by PCR using primers from N22 and untranslated region (UTR) respectively. The established prevalence of TTV tested with N22 and UTR primers according to patients groups was: acute hepatitis of unknown etiology 4 resp. 28 of 37, acute hepatitis B 3 resp. 29 of 38, chronic hepatitis B 11 resp. 41 of 44, chronic hepatitis C 10 resp. 93 of 102, hemodialysis patients 13 resp. 72 of 72, health care workers 0 resp. 27 of 33, control group 8 resp. 83 of 100. Using N22 primers, TTV infection occurred more frequently in chronic hepatitis B group compared with health care workers, if UTR primers were used the group of hemodialysis patients differed significantly from both acute hepatitis groups, health care workers and controls (p < 0.05). From possible risk factors hemodialysis and transfusion count showed notable differences. Bilirubin and aminotransferase levels did not differ between TTV positive and negative groups. No pathogenetic role of TT virus in liver injury was confirmed.

Objective: To investigate the rate of infection caused by Torque teno virus (TTV) in United Arab Emirates (UAEs) healthy population as a pilot study in detecting TTV DNA in 100 healthy blood donors., Methods: We randomly choose a total of 100 healthy blood donors who attended Zayed Military Hospital, Abu Dhabi, UAE from January 20 to May 30, 2005. We carried out a real-time polymerase chain reaction (PCR) test to detect TTV DNA., Results: Real-time for TTV was positive in 75 (75%) donors. Eight (73%) non-UAE donors were TTV positive while 67 (75%) were UAEs. Among these donors, 72 (77%) were males and 3 (50%) were females., Conclusion: Our results demonstrated a high prevalence of TTV in UAE.

SEN virus is a recently discovered DNA virus, and eight genotypes (A to H) were detected by phylogenetic analysis. Genotype D (SENV-D) and H (SENV-H) are thought to be associated in the etiology of post-transfusion hepatitis. Although no strong association was revealed between liver pathology and SEN virus, the viral replication in hepatocytes and transmission by blood transfusions were well characterized. The aim of this study was to investigate the prevalence of SENV-D and SENV-H in blood donors. One hundred serum samples were included in the study which were obtained from blood donors comprised of 98 male and 2 female with a mean age 36.4 years who were enrolled at Mersin University Faculty of Medicine blood center. The DNAs of SENV-D and SENV-H were detected with polymerase chain reaction (PCR), by using D10S/L2AS and C5S/L2AS primers, respectively. SEN virus DNA positivity was detected in 25 of 100 (25%) sera, of which 10 (10%) were positive for SENV-D and 15 (15%) were positive for SENV-H DNA. Although, the number of samples were not sufficient to reflect the general prevalence, our study relatively reveals that asymptomatic carriage rate of these viruses were 25% in our province.

Transfusion-transmitted virus (TTV) has been reported from a number of hemodialysis (HD) units from various countries throughout the world. TTV has been associated with liver diseases, viral hepatitis B, and C. Clinical details and information regarding TTV prevalence from India are insufficient. The prevalence and clinical significance of TTV infection were studied in New Delhi, India in HD patients. Serum samples were derived from 75 patients on maintenance HD, and 75 age- and sex-matched voluntary blood donors were examined for TTV viremia by nested polymerase chain reaction (PCR) using primers derived from UTR (A) region of the TTV genome. The prevalence of TTV DNA in patients on HD (83%) was significantly (p<0.05) higher than in blood donors (43%). Clinical background including the mean age, sex, mean duration of HD, and mean alanine aminotransferase (ALT) levels did not differ significantly between TTV DNA-positive and -negative HD patients. Fifty-four (72%) TTV-positive HD patients and 7 (56%) TTV-negative HD patients had blood transfusion histories (p>0.05). Among TTV-positive patients, Hepatitis B virus (HBV) co-infection was present in 14.2% cases while hepatitis C virus (HCV) co-infection was absent. Persistent elevation of ALT levels was observed in 7(9.3%) HD patients; 3 (43%) of them were TTV positive and 4 (57%) were TTV negative (p>0.05). All 3 TTV-positive patients with elevated ALT levels were co-infected with HBV. Patients with TTV infection alone showed normal ALT levels. Prevalence of TTV infection is high in North Indian patients on maintenance HD. Also, none of the exclusively TTV DNA-positive patients had clinical or biochemical signs of liver disease. TTV seems to spread through parenteral routes. More often, TTV seems to be associated with parenterally transmitted virus HBV, indicating a parenteral mode of TTV transmission. The pathogenicity of TTV remains unclear from the present study.

SEN virus (SEN-V) is a blood-borne, single-stranded, nonenveloped DNA virus. Although its prevalence varies by geographic region, it has been detected in as many as 30 percent of postoperative transfusion recipients, compared to 3 percent of postoperative patients who did not receive transfusions. A significant association has been observed between transfusion volume and the occurrence of SEN-V infection. Transmission by transfusion also has been confirmed by the detection of greater than 99 percent homology between SEN-V in donor and recipient sera. Concurrent infections with SEN-V and hepatitis B virus, hepatitis C virus, or human immunodeficiency virus type 1 have been documented, and these observations probably reflect the blood-borne transmission of these viruses as well as SEN-V. Although SEN-V was discovered as part of a search for causes of posttransfusion hepatitis, there is no firm evidence so far that SEN-V infection either causes hepatitis or worsens the course of coexistent liver disease. Nevertheless, SEN-V appears to be transmitted by transfusion, and further studies may reveal more about its role in the future.

TT virus (TTV) is a naked, single stranded DNA virus, which has been discovered in the serum of a patient with posttransfusion hepatitis of unknown etiology. TTV is widespread in the population, however, the mode of its transmission is unclear. This study was conducted to search for TTV-DNA positivity rates and its relationship with the clinical outcomes of recipients who underwent multiple blood or blood product transfusion, together with healthy children. TTV-DNA was investigated in 52 multitransfused pediatric patients (age range: 3 mnths - 17.5 yrs, mean age: 9.2 +/- 5.7 years) and 18 healthy children (age range: 1 mnth - 16.5 yrs, mean age: 8.1 +/- 4.9 years), by qualitative in-house semi-nested polymerase chain reaction (PCR) with the primers NG059, NG061 and NG063, generated from ORF1 region of the viral genome. TTV-DNA was found positive in 30.8% of multitransfused, and 16.7% of healthy children. The differences of TTV-DNA positivity rates between the multitransfused and control groups, and ALT values between the patients with positive and negative TTV-DNA, were statistically insignificant (p>0.05). As a result, no relationship was detected between TTV positivity and hepatitis, although there was a statistically insignificant increase of TTV-DNA positivity in multitransfused children. However, since the primers of ORF1 N22 region used in our PCR method did not have enough sensitivity for the detection of TTV-DNA, it has been concluded that more sensitive primers such as UTR primers, should be used for more reliable evaluation of the results.

Background: TT virus is prevalent worldwide, but its prevalence and genotype distribution in Central and East-Europe has not been determined. The high prevalence of TTV in multiply-transfused patients points to the importance of a parenteral mode of transmission, but since more than half of the general population is infected other possible routes of transmission must be considered., Methods: In our study, we investigated the epidemiology, transmission and phylogeny of TTV in the Czech Republic. The following groups were selected: a control group of 196 blood donors, 20 patients with hemophilia, 49 intravenous drug users, 100 sex workers, 50 penitentiary prisoners, 208 healthy children aged 1 to 14 years, 54 cord blood samples, 52 patients with non-A-E hepatitis, 74 patients with hepatitis C, and 51 blood donors with increased ALT levels. Primers specific for the non-coding region were used. The genotype distribution was studied in 70 TTV-positive samples., Results: The prevalence rate of TTV among the Czech population was 52.6%. We have shown that TTV is not transmitted prenatally. Children were infected after birth with two peaks: one at the age of two years and the other after the beginning of primary school. Adults have shown a further increase in the TTV prevalence with age. The highest TTV prevalence was found in the group of patients who had received multiple blood transfusions. The TTV prevalence rate in subjects at an increased risk of sexual transmission was not significantly higher than in the general population. Genotypes G2 and G1 were most prevalent among the Czech population, followed by G8 and G3. The subjects positive for markers of HBV and/or HCV infection tested significantly more often TTV DNA positive, which is suggestive of a common route of transmission of these three infections., Conclusions: This study on TTV prevalence, mode of transmission and age-specific prevalence is the most extensive study performed in Central and Eastern Europe. It showed insights into the epidemiology of TTV infection, but failed to associate TTV infection with clinical manifestations.

The TT virus (TTV) was isolated recently from the serum of a patient with post-transfusion hepatitis. TTV infection is widespread in the general population, and its prevalence increases continuously with age. The pathogenic role of TTV in liver disease remains controversial, and the source of transmission is still unclear. We investigated the pathogenicity and epidemiology of TTV infection in infants born to TTV DNA-positive mothers. Enrolled in this study were 22 mother-child pairs testing negative for antibodies to hepatitis B, hepatitis C, and the human immunodeficiency viruses (HIVs). The children were followed for 30 months after birth. Serum TTV DNA was detected by N22-PCR, and the PCR products were cloned and sequenced. The prevalence of TTV infection in children increased with age. Of the 22 children, 13 (59%) became positive for TTV DNA during the follow-up period. Of these 13 children, 6 (46%) had elevated levels of serum alanine aminotransferase (ALT), although the elevations were transient and mild. TTV viremia was not associated significantly with the abnormal ALT levels. Children with TTV viremia developed neither severe liver disease nor fulminant hepatitis. Phylogenetic analysis showed that, in 11 (85%) of the 13 pairs, the mother and child had the same genotype at the first PCR-positive time point. Among those 11 mother-child pairs, 6 (55%) had identical TTV nucleotide sequences. However, the genotype of predominant clones changed in 5 (50%) of 10 children who were positive for TTV DNA at two or more time points during the follow-up period. In conclusion, this study did not provide evidence that TTV infection is related to liver disease in children. Although the main source of TTV infection in children is presumed to be their mothers, transmitted via non-parenteral routes in the course of daily contact, intrafamilial carriers may also be sources of TTV infection.

Ranavirus (family Iridoviridae) is a genus of pathogens of poikilotherms, and some ranaviruses may play a role in widespread mortality of amphibians. Ecology of viral transmission in amphibians is poorly known but can be addressed through experimentation in the laboratory. In this study, we use the Ranavirus frog virus 3 (FV3) as an experimental model for pathogen emergence in naive populations of tadpoles. We simulated emerging disease by exposing tadpoles of the Italian agile frog (Rana latastei), to the North American Ranavirus FV3. We demonstrated that mortality occurred due to viral exposure, exposure of tadpoles to decreasing concentrations of FV3 in the laboratory produced dose-dependent survival rates, and cannibalism of virus-carrying carcasses increased mortality due to FV3. These experiments suggest the potential for ecological mechanisms to affect the level of exposure of tadpoles to Ranavirus and to impact transmission of viral pathogens in aquatic systems.

Nucleopolyhedrovirus (NPV) was tested for vertical transmission in the silkworm, Bombyx mori. Fifth instar larvae were exposed to four different dosages of BmNPV (830, 1300, 1800, and 2000OBs/larva) and a dosage of about 2000OBs/larva was found suitable for obtaining infected adults. Histopathological studies revealed the infection in susceptible tissues and organs initially, and at later stages of infection cycles the spermatocytes and nurse cells in the young oocytes were infected in the larval rudiments of testis and ovary, respectively. The mating of infected females with uninfected males resulted in significant reduction in fecundity (P < 0.01) and hatching of eggs (P < 0.001) due to transovarial transmission of BmNPV. Mating tests of uninfected females and infected males also confirmed venereal transmission as there was a significant reduction in hatching of eggs (P < 0.01). Further, among the F1 hybrid offspring (infected female x uninfected male) that were infected transovarially, larval progeny died at first and second instar stages, whereas those infected venereally developed acute lethal infection late and died by the end of third and fourth instar stage. PCR amplification and sequencing of 473bp of immediate early-1 (ie-1) gene of BmNPV isolated from the viral-infected parent and the F1 offspring confirmed that the viral infection is vertically transmitted to the progeny.

Background: The aim of this study was to evaluate the transfusional transmitted virus (TTV) seroprevalence in asymptomatic HBsAg (+) patients and to assess the influence of TTV on the course of these patients., Methods: Sixty asymptomatic HBV carriers were included and 31 healthy volunteers served as controls. Cases were followed at 6-month intervals for a total duration of 4 years., Results: In the asymptomatic carrier group, 31 patients (51.7%) had a history of surgery and 10 (16.7%) had a history of blood transfusions. TTV-DNA was detected in 45 of these patients (75%). In the control group, 12 patients (38.7%) had a history of surgery and 2 had (6.5%) a history of blood transfusions. TTV-DNA was found in 20 (64.5%) of these subjects. The incidence of TTV-DNA positivity was not significantly different between the two groups (P > 0.05)., Conclusion: In spite of the common occurrence of HBV and TTV, TTV-DNA was also detected in 64.5% of healthy controls. Furthermore, during 4 years of follow up, TTV had no detrimental effects on the course of asymptomatic HBV carriers. These results suggest that the hepatic injury due to TTV is insignificant in this group of patients.

A nested PCR was used to detect TTV DNA in serum from 490 mother-infant pairs to determine the rate of vertical transmission of TTV in Guangzhou City, P.R. China. TTV DNA was positive in 87 mothers and 12 of their babies, giving a prevalence of 17.8% in pregnant women and a mother-to-child transmission rate of 13.8%. In eight mother-infant pairs, TTV DNA nucleotide sequences from the mothers and their corresponding infants were identical. The homology of Guangzhou TTV and Japanese TTV ranged from 85.3%-98.2% at nucleotide level. TTV infection is common in pregnant women in South China and vertical transmission can occur. Guangzhou's and Japanese TTV isolates belong to the same genotype.

Objective: To determine the prevalence and clinical impact of transfusion-transmitted virus (TTV) DNA in patients with chronic liver diseases in the Southeast Anatolia region of Turkey where hepatitis B and C viral infections are endemic., Subjects and Methods: Patients diagnosed with chronic liver disease by clinical, biochemical and histologic means were enrolled in the study. Serum samples of 60 patients (19 males, 41 females) with chronic liver diseases, and of 45 healthy volunteer blood donors as a control group were collected. The chronic liver disease group consisted of 11 patients with hepatitis B, 44 with hepatitis C and 5 with chronic liver disease of unknown etiology. Presence of TTV DNA was investigated by the polymerase chain reaction. Using a scoring system histological grading of inflammation and staging of fibrosis were performed only in the chronic hepatitis C group., Results: TTV DNA was detected in 47 (78%) patients with chronic liver disease and 5 (11%) volunteers in the control group. The difference was statistically significant (p < 0.001). Ten of the 11 (91%) patients with hepatitis B, 32 of 44 (73%) of those with hepatitis C-related chronic liver disease, and 5 of 5 (100%) of the patients with cryptogenic liver disease were positive for TTV DNA., Conclusion: TTV is highly prevalent in patients with chronic liver diseases in Southeast Anatolia, Turkey but no pathogenic effect attributable to TTV infection was detected., (Copyright 2003 S. Karger AG, Basel)

Aim: To determine the prevalences of TTV and HGV infections among blood donors and patients with chronic liver disease in Korea, to investigate the association of TTV and HGV infections with blood transfusion, and to assess the correlation between TTV and HGV viremia and hepatic damage., Methods: A total of 391 serum samples were examined in this study. Samples were obtained from healthy blood donors (n=110), hepatitis B surface antigen (HBsAg)-positive donors (n=112), anti-hepatitis C virus (anti-HCV)-positive donors (n=69), patients with type B chronic liver disease (n=81), and patients with type C chronic liver disease (n=19). TTV DNA was detected using the hemi-nested PCR. HGV RNA was tested using RT-PCR. A history of blood transfusion and serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were also determined., Results: TTV DNA was detected in 8.2 % of healthy blood donors, 16.1 % of HBsAg-positive donors, 20.3 % of anti-HCV-positive donors, 21.0 % of patients with type B chronic liver disease, and 21.1 % of patients with type C chronic liver disease. HGV RNA was detected in 1.8 % of healthy blood donors, 1.8 % of HBsAg-positive donors, 17.4 % of anti-HCV-positive donors, 13.6 % of patients with type B chronic liver disease, and 10.5 % of patients with type C chronic liver disease. The prevalence of TTV and HGV infections in HBV- or HCV-positive donors and patients was significantly higher than in healthy blood donors (P<0.05), except for the detection rate of HGV in HBsAg-positive donors which was the same as for healthy donors. There was a history of transfusion in 66.7 % of TTV DNA-positive patients and 76.9 % of HGV RNA-positive patients (P<0.05). No significant increase in serum ALT and AST was detected in the TTV- or HGV-positive donors and patients., Conclusion: TTV and HGV infections are more frequently found in donors and patients infected with HBV or HCV than in healthy blood donors. However, there is no significant association between TTV or HGV infections and liver injury.

Serum samples from healthy and diseased children were studied for the presence of TTV DNA by nested PCR using primer sets generated from N-22 region and from the untranslated region (UTR) of the viral genome. N-22 positive TTV DNA was detectable in 33 (27%) of 122 healthy children, 47 (73.4%) of 64 polytransfused thalassemic children, 37 (46.3%) of 80 children who received transfusion during cardiac surgery, 8 (42.1%) of 19 non-A to E hepatitis, 10 (33.3%) of 30 HBV carrier children, and 5 (15.6%) of 32 infants with biliary atresia. A much higher prevalence of TTV DNA with rates varying from 78-100% in the above study groups was observed using the UTR primers. For children with N-22 positive TTV DNA, biochemical assessment of isolated TTV viremia in thalassemic children or children transfused during surgery showed no convincing association between raised ALT levels and TTV viremia. Coinfection with TTV in chronic HCV-infected or HBV-infected children did not result in higher peak ALT levels during follow-up, suggesting that TTV has no synergistic pathogenic effect. The phylogenetic analysis of the N-22 positive TTV DNA isolates revealed that most isolates from healthy children, children transfused during surgery, and non-A to E fulminant hepatitis children were type 1 TTV. These results indicate that TTV infection in children was significantly associated with transfusion. TTV infection is highly prevalent in early childhood in Taiwan but plays a minimal role in the induction of hepatitis in children., (Copyright 2003 Wiley-Liss, Inc.)

Background: The major transmission route of Helicobacter pylori, oral-oral or fecal-oral, remains to be established. TT virus (TTV), a recently discovered microbe that is prevalent in healthy persons, is believed to be mainly transmitted by nonparenteral routes. The purpose of this study was to test the hypothesis that these two microorganisms have a common mode of transmission., Methods: We investigated the seroprevalence of H. pylori and TTV in a cross-sectional study of 454 healthy Japanese children from birth to age 15 years, living in five different geographic areas. Determination of H. pylori status was based on the presence of specific serum IgG and IgA antibodies, determined using enzyme immunoassays. TTV DNA was detected and the titer was determined using semiquantitative polymerase chain reaction with heminested primers., Results: The overall prevalences of H. pylori and TTV were 12.2% and 21.6%, respectively. An age-related increase of prevalence was shown for H. pylori ( P < 0.001), but not for TTV ( P = 0.23). Titers of TTV DNA significantly decreased with age (P = 0.02). There were significant geographic differences in TTV prevalence ( P < 0.001), but not in H. pylori seroprevalence (P = 0.33). There was no true correlation between the prevalence of these two organisms (Phi coefficient = -0.02 and P = 0.66)., Conclusions: Although Japanese children frequently acquire both H. pylori and TTV, especially in early childhood, their acquisition appears to be independent.

Background/aims: TT virus is a novel DNA virus that is associated with posttransfusion hepatitis. The prevalence, risk factors, and clinical significance of TT virus infection were evaluated in patients with chronic renal failure who are on hemodialysis., Methodology: Nested polymerase chain reaction was used to test for TT virus DNA in the serum of patients on hemodialysis as well as in healthy controls., Results: TT virus DNA was detected in 15 (20.0%) of the 75 patients on hemodialysis and 10 (13.2%) of the 76 healthy controls (P > 0.05). In chronic renal failure patients on hemodialysis, the prevalence of TT virus did not differ according to the duration of hemodialysis or the amount of blood transfusion. The prevalence of TT virus was higher in IgG anti-hepatitis B core antibody-positive patients than IgG anti-hepatitis B core antibody-negative patients (27.5% vs. 4.2%, P = 0.03). Two (13.3%) of the 15 TT virus-positive and 6 (10.0%) of the 60 TT virus-negative patients showed elevated alanine aminotransferase levels (P > 0.05)., Conclusions: TT virus infection did not occur more frequently in patients on hemodialysis than in healthy controls. No relationship was found between TT virus infection and liver disease. The correlation between TT virus infection and IgG anti-hepatitis B core antibody suggests that TT virus may share some routes of transmission with hepatitis B virus.